GB2324302A - Phenolic acid esterase - Google Patents
Phenolic acid esterase Download PDFInfo
- Publication number
- GB2324302A GB2324302A GB9707540A GB9707540A GB2324302A GB 2324302 A GB2324302 A GB 2324302A GB 9707540 A GB9707540 A GB 9707540A GB 9707540 A GB9707540 A GB 9707540A GB 2324302 A GB2324302 A GB 2324302A
- Authority
- GB
- United Kingdom
- Prior art keywords
- enzyme
- gly
- ggt
- asn
- ala
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 150000007965 phenolic acids Chemical class 0.000 title claims abstract description 34
- 108010013043 Acetylesterase Proteins 0.000 title claims abstract description 31
- 102000004190 Enzymes Human genes 0.000 claims abstract description 121
- 108090000790 Enzymes Proteins 0.000 claims abstract description 121
- 229940088598 enzyme Drugs 0.000 claims abstract description 120
- 230000000694 effects Effects 0.000 claims abstract description 34
- 238000002360 preparation method Methods 0.000 claims abstract description 30
- ZZXREVQVZCYAIE-IGBVEJJSSA-N [(2s,3r,4r)-5-[(2s,3r,4s,5r)-3,5-dihydroxy-2-[(2r,3r,4r)-1,3,4-trihydroxy-5-oxopentan-2-yl]oxyoxan-4-yl]oxy-3,4-dihydroxyoxolan-2-yl]methyl (e)-3-(4-hydroxy-3-methoxyphenyl)prop-2-enoate Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)OC[C@H]2[C@@H]([C@@H](O)C(O[C@@H]3[C@H]([C@H](O[C@H](CO)[C@H](O)[C@@H](O)C=O)OC[C@H]3O)O)O2)O)=C1 ZZXREVQVZCYAIE-IGBVEJJSSA-N 0.000 claims abstract description 20
- 239000000758 substrate Substances 0.000 claims abstract description 20
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 18
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 14
- 238000004519 manufacturing process Methods 0.000 claims abstract description 14
- 239000000872 buffer Substances 0.000 claims abstract description 10
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 9
- 239000005017 polysaccharide Substances 0.000 claims abstract description 9
- 150000004804 polysaccharides Chemical class 0.000 claims abstract description 9
- 230000000593 degrading effect Effects 0.000 claims abstract description 8
- 241000422444 Piromyces equi Species 0.000 claims abstract description 7
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims abstract description 6
- 229910000397 disodium phosphate Inorganic materials 0.000 claims abstract description 6
- 235000019800 disodium phosphate Nutrition 0.000 claims abstract description 6
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims abstract description 5
- -1 arabinase Proteins 0.000 claims abstract description 5
- 239000013598 vector Substances 0.000 claims abstract description 5
- 108010059892 Cellulase Proteins 0.000 claims abstract description 4
- 239000000463 material Substances 0.000 claims abstract description 4
- 235000009048 phenolic acids Nutrition 0.000 claims abstract description 4
- 235000000346 sugar Nutrition 0.000 claims abstract description 4
- 239000002699 waste material Substances 0.000 claims abstract description 4
- 108010011619 6-Phytase Proteins 0.000 claims abstract description 3
- 239000004382 Amylase Substances 0.000 claims abstract description 3
- 102000013142 Amylases Human genes 0.000 claims abstract description 3
- 108010065511 Amylases Proteins 0.000 claims abstract description 3
- 102100032487 Beta-mannosidase Human genes 0.000 claims abstract description 3
- 108090000604 Hydrolases Proteins 0.000 claims abstract description 3
- 102000004157 Hydrolases Human genes 0.000 claims abstract description 3
- 108090001060 Lipase Proteins 0.000 claims abstract description 3
- 102000004882 Lipase Human genes 0.000 claims abstract description 3
- 239000004367 Lipase Substances 0.000 claims abstract description 3
- 241001465754 Metazoa Species 0.000 claims abstract description 3
- 102100036617 Monoacylglycerol lipase ABHD2 Human genes 0.000 claims abstract description 3
- 108091005804 Peptidases Proteins 0.000 claims abstract description 3
- 108010059820 Polygalacturonase Proteins 0.000 claims abstract description 3
- 239000004365 Protease Substances 0.000 claims abstract description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 3
- 108010093941 acetylxylan esterase Proteins 0.000 claims abstract description 3
- 235000019418 amylase Nutrition 0.000 claims abstract description 3
- 108010055059 beta-Mannosidase Proteins 0.000 claims abstract description 3
- 108010080434 cephalosporin-C deacetylase Proteins 0.000 claims abstract description 3
- 244000038559 crop plants Species 0.000 claims abstract description 3
- 235000019621 digestibility Nutrition 0.000 claims abstract description 3
- 108010093305 exopolygalacturonase Proteins 0.000 claims abstract description 3
- 125000003147 glycosyl group Chemical group 0.000 claims abstract description 3
- 235000019421 lipase Nutrition 0.000 claims abstract description 3
- 244000144972 livestock Species 0.000 claims abstract description 3
- 229940085127 phytase Drugs 0.000 claims abstract description 3
- 150000008163 sugars Chemical class 0.000 claims abstract description 3
- 108020004414 DNA Proteins 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 12
- 102000053602 DNA Human genes 0.000 claims description 9
- 108010041969 feruloyl esterase Proteins 0.000 claims description 7
- 239000001913 cellulose Substances 0.000 claims description 6
- 229920002678 cellulose Polymers 0.000 claims description 6
- 239000003674 animal food additive Substances 0.000 claims description 4
- 235000013305 food Nutrition 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 241000193632 Piromyces sp. Species 0.000 claims description 2
- 108010060309 Glucuronidase Proteins 0.000 claims 1
- 102000053187 Glucuronidase Human genes 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 abstract description 6
- 229940114124 ferulic acid Drugs 0.000 abstract description 6
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 abstract description 6
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 abstract description 6
- 235000001785 ferulic acid Nutrition 0.000 abstract description 6
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 abstract description 6
- 241000235379 Piromyces Species 0.000 abstract 1
- 102000005840 alpha-Galactosidase Human genes 0.000 abstract 1
- 108010030291 alpha-Galactosidase Proteins 0.000 abstract 1
- 108010084650 alpha-N-arabinofuranosidase Proteins 0.000 abstract 1
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 46
- RLMISHABBKUNFO-WHFBIAKZSA-N Ala-Ala-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O RLMISHABBKUNFO-WHFBIAKZSA-N 0.000 description 32
- 108010078144 glutaminyl-glycine Proteins 0.000 description 28
- 108010090461 DFG peptide Proteins 0.000 description 20
- WJZLEENECIOOSA-WDSKDSINSA-N Gly-Asn-Gln Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)O WJZLEENECIOOSA-WDSKDSINSA-N 0.000 description 18
- 241000196324 Embryophyta Species 0.000 description 15
- KMSGYZQRXPUKGI-BYPYZUCNSA-N Gly-Gly-Asn Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(N)=O KMSGYZQRXPUKGI-BYPYZUCNSA-N 0.000 description 14
- 108010038745 tryptophylglycine Proteins 0.000 description 14
- BYXHQQCXAJARLQ-ZLUOBGJFSA-N Ala-Ala-Ala Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O BYXHQQCXAJARLQ-ZLUOBGJFSA-N 0.000 description 13
- KVNOBVKRBOYSIV-SZMVWBNQSA-N Met-Pro-Trp Chemical compound CSCC[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O)N KVNOBVKRBOYSIV-SZMVWBNQSA-N 0.000 description 12
- QJHOOKBAHRJPPX-QWRGUYRKSA-N Asp-Phe-Gly Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 QJHOOKBAHRJPPX-QWRGUYRKSA-N 0.000 description 10
- RPLLQZBOVIVGMX-QWRGUYRKSA-N Gly-Asp-Phe Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O RPLLQZBOVIVGMX-QWRGUYRKSA-N 0.000 description 10
- QPCVIQJVRGXUSA-LURJTMIESA-N Gly-Gly-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)CNC(=O)CN QPCVIQJVRGXUSA-LURJTMIESA-N 0.000 description 10
- 210000002421 cell wall Anatomy 0.000 description 10
- VJTWLBMESLDOMK-WDSKDSINSA-N Asn-Gln-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O VJTWLBMESLDOMK-WDSKDSINSA-N 0.000 description 8
- LPAJOCKCPRZEAG-MNXVOIDGSA-N Lys-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCCCN LPAJOCKCPRZEAG-MNXVOIDGSA-N 0.000 description 8
- 150000001413 amino acids Chemical group 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 229920001221 xylan Polymers 0.000 description 7
- 150000004823 xylans Chemical class 0.000 description 7
- NITWSHWHQAQBAW-QPJJXVBHSA-N (E)-4-coumaric acid methyl ester Chemical compound COC(=O)\C=C\C1=CC=C(O)C=C1 NITWSHWHQAQBAW-QPJJXVBHSA-N 0.000 description 6
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 6
- RGNMNWULPAYDAH-JSGCOSHPSA-N Gln-Trp-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCC(=O)N)N RGNMNWULPAYDAH-JSGCOSHPSA-N 0.000 description 6
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 108010077515 glycylproline Proteins 0.000 description 6
- 108010051242 phenylalanylserine Proteins 0.000 description 6
- AUJXJFHANFIVKH-GQCTYLIASA-N trans-methylferulate Chemical compound COC(=O)\C=C\C1=CC=C(O)C(OC)=C1 AUJXJFHANFIVKH-GQCTYLIASA-N 0.000 description 6
- 229920005610 lignin Polymers 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- MIPWEZAIMPYQST-FXQIFTODSA-N Ala-Cys-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O MIPWEZAIMPYQST-FXQIFTODSA-N 0.000 description 4
- PAIHPOGPJVUFJY-WDSKDSINSA-N Ala-Glu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PAIHPOGPJVUFJY-WDSKDSINSA-N 0.000 description 4
- QHASENCZLDHBGX-ONGXEEELSA-N Ala-Gly-Phe Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QHASENCZLDHBGX-ONGXEEELSA-N 0.000 description 4
- HHRAXZAYZFFRAM-CIUDSAMLSA-N Ala-Leu-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O HHRAXZAYZFFRAM-CIUDSAMLSA-N 0.000 description 4
- WUQXMTITJLFXAU-JIOCBJNQSA-N Asn-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)N)N)O WUQXMTITJLFXAU-JIOCBJNQSA-N 0.000 description 4
- PXLNPFOJZQMXAT-BYULHYEWSA-N Asp-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O PXLNPFOJZQMXAT-BYULHYEWSA-N 0.000 description 4
- DTNUIAJCPRMNBT-WHFBIAKZSA-N Asp-Gly-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](C)C(O)=O DTNUIAJCPRMNBT-WHFBIAKZSA-N 0.000 description 4
- PZXPWHFYZXTFBI-YUMQZZPRSA-N Asp-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(O)=O PZXPWHFYZXTFBI-YUMQZZPRSA-N 0.000 description 4
- JNNVNVRBYUJYGS-CIUDSAMLSA-N Asp-Leu-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O JNNVNVRBYUJYGS-CIUDSAMLSA-N 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- XSBGUANSZDGULP-IUCAKERBSA-N Gln-Gly-Lys Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CCCCN)C(O)=O XSBGUANSZDGULP-IUCAKERBSA-N 0.000 description 4
- SOEXCCGNHQBFPV-DLOVCJGASA-N Gln-Val-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(O)=O SOEXCCGNHQBFPV-DLOVCJGASA-N 0.000 description 4
- CIMULJZTTOBOPN-WHFBIAKZSA-N Gly-Asn-Asn Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CIMULJZTTOBOPN-WHFBIAKZSA-N 0.000 description 4
- YIFUFYZELCMPJP-YUMQZZPRSA-N Gly-Leu-Cys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(O)=O YIFUFYZELCMPJP-YUMQZZPRSA-N 0.000 description 4
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 4
- VLIJYPMATZSOLL-YUMQZZPRSA-N Gly-Lys-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN VLIJYPMATZSOLL-YUMQZZPRSA-N 0.000 description 4
- WNZOCXUOGVYYBJ-CDMKHQONSA-N Gly-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)CN)O WNZOCXUOGVYYBJ-CDMKHQONSA-N 0.000 description 4
- CSMYMGFCEJWALV-WDSKDSINSA-N Gly-Ser-Gln Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(N)=O CSMYMGFCEJWALV-WDSKDSINSA-N 0.000 description 4
- UAVQIQOOBXFKRC-BYULHYEWSA-N Ile-Asn-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O UAVQIQOOBXFKRC-BYULHYEWSA-N 0.000 description 4
- TWYOYAKMLHWMOJ-ZPFDUUQYSA-N Ile-Leu-Asn Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O TWYOYAKMLHWMOJ-ZPFDUUQYSA-N 0.000 description 4
- FJWALBCCVIHZBS-QXEWZRGKSA-N Ile-Met-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)O)N FJWALBCCVIHZBS-QXEWZRGKSA-N 0.000 description 4
- NJGXXYLPDMMFJB-XUXIUFHCSA-N Ile-Val-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N NJGXXYLPDMMFJB-XUXIUFHCSA-N 0.000 description 4
- 108010065920 Insulin Lispro Proteins 0.000 description 4
- ZRLUISBDKUWAIZ-CIUDSAMLSA-N Leu-Ala-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O ZRLUISBDKUWAIZ-CIUDSAMLSA-N 0.000 description 4
- USLNHQZCDQJBOV-ZPFDUUQYSA-N Leu-Ile-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O USLNHQZCDQJBOV-ZPFDUUQYSA-N 0.000 description 4
- VTJUNIYRYIAIHF-IUCAKERBSA-N Leu-Pro Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(O)=O VTJUNIYRYIAIHF-IUCAKERBSA-N 0.000 description 4
- ABHIXYDMILIUKV-CIUDSAMLSA-N Lys-Asn-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ABHIXYDMILIUKV-CIUDSAMLSA-N 0.000 description 4
- ISHNZELVUVPCHY-ZETCQYMHSA-N Lys-Gly-Gly Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)NCC(O)=O ISHNZELVUVPCHY-ZETCQYMHSA-N 0.000 description 4
- GHKXHCMRAUYLBS-CIUDSAMLSA-N Lys-Ser-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O GHKXHCMRAUYLBS-CIUDSAMLSA-N 0.000 description 4
- RPWTZTBIFGENIA-VOAKCMCISA-N Lys-Thr-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RPWTZTBIFGENIA-VOAKCMCISA-N 0.000 description 4
- MHQXIBRPDKXDGZ-ZFWWWQNUSA-N Met-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)[C@@H](N)CCSC)C(O)=O)=CNC2=C1 MHQXIBRPDKXDGZ-ZFWWWQNUSA-N 0.000 description 4
- PESQCPHRXOFIPX-UHFFFAOYSA-N N-L-methionyl-L-tyrosine Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 PESQCPHRXOFIPX-UHFFFAOYSA-N 0.000 description 4
- 108010047562 NGR peptide Proteins 0.000 description 4
- CGOMLCQJEMWMCE-STQMWFEESA-N Phe-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 CGOMLCQJEMWMCE-STQMWFEESA-N 0.000 description 4
- NAXPHWZXEXNDIW-JTQLQIEISA-N Phe-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 NAXPHWZXEXNDIW-JTQLQIEISA-N 0.000 description 4
- WLYPRKLMRIYGPP-JYJNAYRXSA-N Phe-Lys-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 WLYPRKLMRIYGPP-JYJNAYRXSA-N 0.000 description 4
- NJJBATPLUQHRBM-IHRRRGAJSA-N Phe-Pro-Ser Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)N)C(=O)N[C@@H](CO)C(=O)O NJJBATPLUQHRBM-IHRRRGAJSA-N 0.000 description 4
- WFLWKEUBTSOFMP-FXQIFTODSA-N Pro-Cys-Cys Chemical compound OC(=O)[C@H](CS)NC(=O)[C@H](CS)NC(=O)[C@@H]1CCCN1 WFLWKEUBTSOFMP-FXQIFTODSA-N 0.000 description 4
- INDVYIOKMXFQFM-SRVKXCTJSA-N Pro-Lys-Gln Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(=O)N)C(=O)O INDVYIOKMXFQFM-SRVKXCTJSA-N 0.000 description 4
- IDQFQFVEWMWRQQ-DLOVCJGASA-N Ser-Ala-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O IDQFQFVEWMWRQQ-DLOVCJGASA-N 0.000 description 4
- VXYQOFXBIXKPCX-BQBZGAKWSA-N Ser-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CO)N VXYQOFXBIXKPCX-BQBZGAKWSA-N 0.000 description 4
- ZUXQFMVPAYGPFJ-JXUBOQSCSA-N Thr-Ala-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN ZUXQFMVPAYGPFJ-JXUBOQSCSA-N 0.000 description 4
- SKHPKKYKDYULDH-HJGDQZAQSA-N Thr-Asn-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O SKHPKKYKDYULDH-HJGDQZAQSA-N 0.000 description 4
- LMMDEZPNUTZJAY-GCJQMDKQSA-N Thr-Asp-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O LMMDEZPNUTZJAY-GCJQMDKQSA-N 0.000 description 4
- WLDUCKSCDRIVLJ-NUMRIWBASA-N Thr-Gln-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O WLDUCKSCDRIVLJ-NUMRIWBASA-N 0.000 description 4
- IQPWNQRRAJHOKV-KATARQTJSA-N Thr-Ser-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN IQPWNQRRAJHOKV-KATARQTJSA-N 0.000 description 4
- HYVLNORXQGKONN-NUTKFTJISA-N Trp-Ala-Lys Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O)=CNC2=C1 HYVLNORXQGKONN-NUTKFTJISA-N 0.000 description 4
- STTVVMWQKDOKAM-YESZJQIVSA-N Tyr-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC3=CC=C(C=C3)O)N)C(=O)O STTVVMWQKDOKAM-YESZJQIVSA-N 0.000 description 4
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 4
- CPGJELLYDQEDRK-NAKRPEOUSA-N Val-Ile-Ala Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](C)C(O)=O CPGJELLYDQEDRK-NAKRPEOUSA-N 0.000 description 4
- VPGCVZRRBYOGCD-AVGNSLFASA-N Val-Lys-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O VPGCVZRRBYOGCD-AVGNSLFASA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 108010041407 alanylaspartic acid Proteins 0.000 description 4
- 108010038850 arginyl-isoleucyl-tyrosine Proteins 0.000 description 4
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 4
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 4
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 4
- 108010089804 glycyl-threonine Proteins 0.000 description 4
- 108010037850 glycylvaline Proteins 0.000 description 4
- 108010078274 isoleucylvaline Proteins 0.000 description 4
- 108010034529 leucyl-lysine Proteins 0.000 description 4
- 108010057821 leucylproline Proteins 0.000 description 4
- 108010025153 lysyl-alanyl-alanine Proteins 0.000 description 4
- 108010076718 lysyl-glutamyl-tryptophan Proteins 0.000 description 4
- 108010063431 methionyl-aspartyl-glycine Proteins 0.000 description 4
- 108010005942 methionylglycine Proteins 0.000 description 4
- 108010029020 prolylglycine Proteins 0.000 description 4
- 108010061238 threonyl-glycine Proteins 0.000 description 4
- 108010072986 threonyl-seryl-lysine Proteins 0.000 description 4
- 230000009261 transgenic effect Effects 0.000 description 4
- 108010005834 tyrosyl-alanyl-glycine Proteins 0.000 description 4
- 108090000371 Esterases Proteins 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- 229920002488 Hemicellulose Polymers 0.000 description 3
- 239000007993 MOPS buffer Substances 0.000 description 3
- JTLOUXXZZFFBBW-UHFFFAOYSA-N isoferulic acid methyl ester Natural products COC(=O)C=CC1=CC=C(OC)C(O)=C1 JTLOUXXZZFFBBW-UHFFFAOYSA-N 0.000 description 3
- AUJXJFHANFIVKH-UHFFFAOYSA-N methyl cis-ferulate Natural products COC(=O)C=CC1=CC=C(O)C(OC)=C1 AUJXJFHANFIVKH-UHFFFAOYSA-N 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- YBADLXQNJCMBKR-UHFFFAOYSA-M (4-nitrophenyl)acetate Chemical compound [O-]C(=O)CC1=CC=C([N+]([O-])=O)C=C1 YBADLXQNJCMBKR-UHFFFAOYSA-M 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- DVLFYONBTKHTER-UHFFFAOYSA-N 3-(N-morpholino)propanesulfonic acid Chemical compound OS(=O)(=O)CCCN1CCOCC1 DVLFYONBTKHTER-UHFFFAOYSA-N 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- WQVYAWIMAWTGMW-ZLUOBGJFSA-N Ala-Asp-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N WQVYAWIMAWTGMW-ZLUOBGJFSA-N 0.000 description 2
- IFTVANMRTIHKML-WDSKDSINSA-N Ala-Gln-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O IFTVANMRTIHKML-WDSKDSINSA-N 0.000 description 2
- LJFNNUBZSZCZFN-WHFBIAKZSA-N Ala-Gly-Cys Chemical compound N[C@@H](C)C(=O)NCC(=O)N[C@@H](CS)C(=O)O LJFNNUBZSZCZFN-WHFBIAKZSA-N 0.000 description 2
- VHVVPYOJIIQCKS-QEJZJMRPSA-N Ala-Leu-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 VHVVPYOJIIQCKS-QEJZJMRPSA-N 0.000 description 2
- ADSGHMXEAZJJNF-DCAQKATOSA-N Ala-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N ADSGHMXEAZJJNF-DCAQKATOSA-N 0.000 description 2
- YYAVDNKUWLAFCV-ACZMJKKPSA-N Ala-Ser-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O YYAVDNKUWLAFCV-ACZMJKKPSA-N 0.000 description 2
- KWTVWJPNHAOREN-IHRRRGAJSA-N Arg-Asn-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O KWTVWJPNHAOREN-IHRRRGAJSA-N 0.000 description 2
- FNXCAFKDGBROCU-STECZYCISA-N Arg-Ile-Tyr Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 FNXCAFKDGBROCU-STECZYCISA-N 0.000 description 2
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 2
- PAPSMOYMQDWIOR-AVGNSLFASA-N Arg-Lys-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O PAPSMOYMQDWIOR-AVGNSLFASA-N 0.000 description 2
- YFHATWYGAAXQCF-JYJNAYRXSA-N Arg-Pro-Phe Chemical compound NC(N)=NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YFHATWYGAAXQCF-JYJNAYRXSA-N 0.000 description 2
- LEFKSBYHUGUWLP-ACZMJKKPSA-N Asn-Ala-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LEFKSBYHUGUWLP-ACZMJKKPSA-N 0.000 description 2
- KSBHCUSPLWRVEK-ZLUOBGJFSA-N Asn-Asn-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O KSBHCUSPLWRVEK-ZLUOBGJFSA-N 0.000 description 2
- KXFCBAHYSLJCCY-ZLUOBGJFSA-N Asn-Asn-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O KXFCBAHYSLJCCY-ZLUOBGJFSA-N 0.000 description 2
- PLVAAIPKSGUXDV-WHFBIAKZSA-N Asn-Gly-Cys Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N)C(=O)N PLVAAIPKSGUXDV-WHFBIAKZSA-N 0.000 description 2
- PTSDPWIHOYMRGR-UGYAYLCHSA-N Asn-Ile-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O PTSDPWIHOYMRGR-UGYAYLCHSA-N 0.000 description 2
- MVXJBVVLACEGCG-PCBIJLKTSA-N Asn-Phe-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MVXJBVVLACEGCG-PCBIJLKTSA-N 0.000 description 2
- HPBNLFLSSQDFQW-WHFBIAKZSA-N Asn-Ser-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O HPBNLFLSSQDFQW-WHFBIAKZSA-N 0.000 description 2
- JPPLRQVZMZFOSX-UWJYBYFXSA-N Asn-Tyr-Ala Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 JPPLRQVZMZFOSX-UWJYBYFXSA-N 0.000 description 2
- ZAESWDKAMDVHLL-RCOVLWMOSA-N Asn-Val-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O ZAESWDKAMDVHLL-RCOVLWMOSA-N 0.000 description 2
- WQAOZCVOOYUWKG-LSJOCFKGSA-N Asn-Val-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CC(=O)N)N WQAOZCVOOYUWKG-LSJOCFKGSA-N 0.000 description 2
- NURJSGZGBVJFAD-ZLUOBGJFSA-N Asp-Cys-Ser Chemical compound C([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(=O)O)N)C(=O)O NURJSGZGBVJFAD-ZLUOBGJFSA-N 0.000 description 2
- WBDWQKRLTVCDSY-WHFBIAKZSA-N Asp-Gly-Asp Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O WBDWQKRLTVCDSY-WHFBIAKZSA-N 0.000 description 2
- LIVXPXUVXFRWNY-CIUDSAMLSA-N Asp-Lys-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O LIVXPXUVXFRWNY-CIUDSAMLSA-N 0.000 description 2
- GPPIDDWYKJPRES-YDHLFZDLSA-N Asp-Phe-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GPPIDDWYKJPRES-YDHLFZDLSA-N 0.000 description 2
- UAXIKORUDGGIGA-DCAQKATOSA-N Asp-Pro-Lys Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC(=O)O)N)C(=O)N[C@@H](CCCCN)C(=O)O UAXIKORUDGGIGA-DCAQKATOSA-N 0.000 description 2
- WAEDSQFVZJUHLI-BYULHYEWSA-N Asp-Val-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O WAEDSQFVZJUHLI-BYULHYEWSA-N 0.000 description 2
- 241000228212 Aspergillus Species 0.000 description 2
- 101001049242 Aspergillus niger Feruloyl esterase B Proteins 0.000 description 2
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 2
- 101100505161 Caenorhabditis elegans mel-32 gene Proteins 0.000 description 2
- SDXQKJAWASHMIZ-CIUDSAMLSA-N Cys-Glu-Met Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O SDXQKJAWASHMIZ-CIUDSAMLSA-N 0.000 description 2
- NXQCSPVUPLUTJH-WHFBIAKZSA-N Cys-Ser-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O NXQCSPVUPLUTJH-WHFBIAKZSA-N 0.000 description 2
- ALNKNYKSZPSLBD-ZDLURKLDSA-N Cys-Thr-Gly Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O ALNKNYKSZPSLBD-ZDLURKLDSA-N 0.000 description 2
- OVQXQLWWJSNYFV-XEGUGMAKSA-N Gln-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCC(N)=O)C)C(O)=O)=CNC2=C1 OVQXQLWWJSNYFV-XEGUGMAKSA-N 0.000 description 2
- JEFZIKRIDLHOIF-BYPYZUCNSA-N Gln-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(O)=O JEFZIKRIDLHOIF-BYPYZUCNSA-N 0.000 description 2
- DRNMNLKUUKKPIA-HTUGSXCWSA-N Gln-Phe-Thr Chemical compound C[C@@H](O)[C@H](NC(=O)[C@H](Cc1ccccc1)NC(=O)[C@@H](N)CCC(N)=O)C(O)=O DRNMNLKUUKKPIA-HTUGSXCWSA-N 0.000 description 2
- DITJVHONFRJKJW-BPUTZDHNSA-N Gln-Trp-Glu Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N DITJVHONFRJKJW-BPUTZDHNSA-N 0.000 description 2
- NKSGKPWXSWBRRX-ACZMJKKPSA-N Glu-Asn-Cys Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CS)C(=O)O)N NKSGKPWXSWBRRX-ACZMJKKPSA-N 0.000 description 2
- BUZMZDDKFCSKOT-CIUDSAMLSA-N Glu-Glu-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O BUZMZDDKFCSKOT-CIUDSAMLSA-N 0.000 description 2
- ZTVGZOIBLRPQNR-KKUMJFAQSA-N Glu-Met-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZTVGZOIBLRPQNR-KKUMJFAQSA-N 0.000 description 2
- VXEFAWJTFAUDJK-AVGNSLFASA-N Glu-Tyr-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O VXEFAWJTFAUDJK-AVGNSLFASA-N 0.000 description 2
- GZUKEVBTYNNUQF-WDSKDSINSA-N Gly-Ala-Gln Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(O)=O GZUKEVBTYNNUQF-WDSKDSINSA-N 0.000 description 2
- KFMBRBPXHVMDFN-UWVGGRQHSA-N Gly-Arg-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCNC(N)=N KFMBRBPXHVMDFN-UWVGGRQHSA-N 0.000 description 2
- MHHUEAIBJZWDBH-YUMQZZPRSA-N Gly-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)CN MHHUEAIBJZWDBH-YUMQZZPRSA-N 0.000 description 2
- JPWIMMUNWUKOAD-STQMWFEESA-N Gly-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)CN JPWIMMUNWUKOAD-STQMWFEESA-N 0.000 description 2
- MQVNVZUEPUIAFA-WDSKDSINSA-N Gly-Cys-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)CN MQVNVZUEPUIAFA-WDSKDSINSA-N 0.000 description 2
- CQZDZKRHFWJXDF-WDSKDSINSA-N Gly-Gln-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CN CQZDZKRHFWJXDF-WDSKDSINSA-N 0.000 description 2
- PDAWDNVHMUKWJR-ZETCQYMHSA-N Gly-Gly-His Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC1=CNC=N1 PDAWDNVHMUKWJR-ZETCQYMHSA-N 0.000 description 2
- IFHJOBKVXBESRE-YUMQZZPRSA-N Gly-Met-Gln Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)CN IFHJOBKVXBESRE-YUMQZZPRSA-N 0.000 description 2
- OCPPBNKYGYSLOE-IUCAKERBSA-N Gly-Pro-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN OCPPBNKYGYSLOE-IUCAKERBSA-N 0.000 description 2
- SYOJVRNQCXYEOV-XVKPBYJWSA-N Gly-Val-Glu Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SYOJVRNQCXYEOV-XVKPBYJWSA-N 0.000 description 2
- 101001072579 Homo sapiens Protein PAXX Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- LKACSKJPTFSBHR-MNXVOIDGSA-N Ile-Gln-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N LKACSKJPTFSBHR-MNXVOIDGSA-N 0.000 description 2
- WXLYNEHOGRYNFU-URLPEUOOSA-N Ile-Thr-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N WXLYNEHOGRYNFU-URLPEUOOSA-N 0.000 description 2
- DTPGSUQHUMELQB-GVARAGBVSA-N Ile-Tyr-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 DTPGSUQHUMELQB-GVARAGBVSA-N 0.000 description 2
- YJRSIJZUIUANHO-NAKRPEOUSA-N Ile-Val-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(=O)O)N YJRSIJZUIUANHO-NAKRPEOUSA-N 0.000 description 2
- MVHXGBZUJLWZOH-BJDJZHNGSA-N Leu-Ser-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MVHXGBZUJLWZOH-BJDJZHNGSA-N 0.000 description 2
- ILDSIMPXNFWKLH-KATARQTJSA-N Leu-Thr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ILDSIMPXNFWKLH-KATARQTJSA-N 0.000 description 2
- UWKNTTJNVSYXPC-CIUDSAMLSA-N Lys-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN UWKNTTJNVSYXPC-CIUDSAMLSA-N 0.000 description 2
- PGBPWPTUOSCNLE-JYJNAYRXSA-N Lys-Gln-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N PGBPWPTUOSCNLE-JYJNAYRXSA-N 0.000 description 2
- BOJYMMBYBNOOGG-DCAQKATOSA-N Lys-Pro-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O BOJYMMBYBNOOGG-DCAQKATOSA-N 0.000 description 2
- ACYHZNZHIZWLQF-BQBZGAKWSA-N Met-Asn-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O ACYHZNZHIZWLQF-BQBZGAKWSA-N 0.000 description 2
- FJVJLMZUIGMFFU-BQBZGAKWSA-N Met-Asp-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O FJVJLMZUIGMFFU-BQBZGAKWSA-N 0.000 description 2
- IECZNARPMKQGJC-XIRDDKMYSA-N Met-Gln-Trp Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N IECZNARPMKQGJC-XIRDDKMYSA-N 0.000 description 2
- IUYCGMNKIZDRQI-BQBZGAKWSA-N Met-Gly-Ala Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O IUYCGMNKIZDRQI-BQBZGAKWSA-N 0.000 description 2
- LCPUWQLULVXROY-RHYQMDGZSA-N Met-Lys-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LCPUWQLULVXROY-RHYQMDGZSA-N 0.000 description 2
- RMLWDZINJUDMEB-IHRRRGAJSA-N Met-Tyr-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N RMLWDZINJUDMEB-IHRRRGAJSA-N 0.000 description 2
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 2
- OAOLATANIHTNCZ-IHRRRGAJSA-N Phe-Met-Asp Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N OAOLATANIHTNCZ-IHRRRGAJSA-N 0.000 description 2
- UXQFHEKRGHYJRA-STQMWFEESA-N Phe-Met-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O UXQFHEKRGHYJRA-STQMWFEESA-N 0.000 description 2
- LTAWNJXSRUCFAN-UNQGMJICSA-N Phe-Thr-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O LTAWNJXSRUCFAN-UNQGMJICSA-N 0.000 description 2
- 241000209504 Poaceae Species 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- VXCHGLYSIOOZIS-GUBZILKMSA-N Pro-Ala-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 VXCHGLYSIOOZIS-GUBZILKMSA-N 0.000 description 2
- KLSOMAFWRISSNI-OSUNSFLBSA-N Pro-Ile-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]1CCCN1 KLSOMAFWRISSNI-OSUNSFLBSA-N 0.000 description 2
- JUJCUYWRJMFJJF-AVGNSLFASA-N Pro-Lys-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H]1CCCN1 JUJCUYWRJMFJJF-AVGNSLFASA-N 0.000 description 2
- WIPAMEKBSHNFQE-IUCAKERBSA-N Pro-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@@H]1CCCN1 WIPAMEKBSHNFQE-IUCAKERBSA-N 0.000 description 2
- ZVEQWRWMRFIVSD-HRCADAONSA-N Pro-Phe-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N3CCC[C@@H]3C(=O)O ZVEQWRWMRFIVSD-HRCADAONSA-N 0.000 description 2
- CXGLFEOYCJFKPR-RCWTZXSCSA-N Pro-Thr-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O CXGLFEOYCJFKPR-RCWTZXSCSA-N 0.000 description 2
- DMNANGOFEUVBRV-GJZGRUSLSA-N Pro-Trp-Gly Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)O)C(=O)[C@@H]1CCCN1 DMNANGOFEUVBRV-GJZGRUSLSA-N 0.000 description 2
- 102100036663 Protein PAXX Human genes 0.000 description 2
- 241000235070 Saccharomyces Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 2
- ICHZYBVODUVUKN-SRVKXCTJSA-N Ser-Asn-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ICHZYBVODUVUKN-SRVKXCTJSA-N 0.000 description 2
- MMAPOBOTRUVNKJ-ZLUOBGJFSA-N Ser-Asp-Ser Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CO)N)C(=O)O MMAPOBOTRUVNKJ-ZLUOBGJFSA-N 0.000 description 2
- KDGARKCAKHBEDB-NKWVEPMBSA-N Ser-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CO)N)C(=O)O KDGARKCAKHBEDB-NKWVEPMBSA-N 0.000 description 2
- DOSZISJPMCYEHT-NAKRPEOUSA-N Ser-Ile-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O DOSZISJPMCYEHT-NAKRPEOUSA-N 0.000 description 2
- ZUUDNCOCILSYAM-KKHAAJSZSA-N Thr-Asp-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O ZUUDNCOCILSYAM-KKHAAJSZSA-N 0.000 description 2
- AQAMPXBRJJWPNI-JHEQGTHGSA-N Thr-Gly-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AQAMPXBRJJWPNI-JHEQGTHGSA-N 0.000 description 2
- PAXANSWUSVPFNK-IUKAMOBKSA-N Thr-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N PAXANSWUSVPFNK-IUKAMOBKSA-N 0.000 description 2
- KPMIQCXJDVKWKO-IFFSRLJSSA-N Thr-Val-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KPMIQCXJDVKWKO-IFFSRLJSSA-N 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- XZSJDSBPEJBEFZ-QRTARXTBSA-N Trp-Asn-Val Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O XZSJDSBPEJBEFZ-QRTARXTBSA-N 0.000 description 2
- PMIJXCLOQFMOKZ-BPUTZDHNSA-N Trp-Asp-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N PMIJXCLOQFMOKZ-BPUTZDHNSA-N 0.000 description 2
- WQYPAGQDXAJNED-AAEUAGOBSA-N Trp-Cys-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CS)C(=O)NCC(=O)O)N WQYPAGQDXAJNED-AAEUAGOBSA-N 0.000 description 2
- UDCHKDYNMRJYMI-QEJZJMRPSA-N Trp-Glu-Ser Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O UDCHKDYNMRJYMI-QEJZJMRPSA-N 0.000 description 2
- FNOQJVHFVLVMOS-AAEUAGOBSA-N Trp-Gly-Asn Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)N)C(=O)O)N FNOQJVHFVLVMOS-AAEUAGOBSA-N 0.000 description 2
- BEWOXKJJMBKRQL-AAEUAGOBSA-N Trp-Gly-Asp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N BEWOXKJJMBKRQL-AAEUAGOBSA-N 0.000 description 2
- KXIQQAWIPDDVOE-BPUTZDHNSA-N Trp-Pro-Cys Chemical compound O=C([C@H](CC=1C2=CC=CC=C2NC=1)N)N1CCC[C@H]1C(=O)N[C@@H](CS)C(O)=O KXIQQAWIPDDVOE-BPUTZDHNSA-N 0.000 description 2
- UIRPULWLRODAEQ-QEJZJMRPSA-N Trp-Ser-Glu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 UIRPULWLRODAEQ-QEJZJMRPSA-N 0.000 description 2
- GFHYISDTIWZUSU-QWRGUYRKSA-N Tyr-Asn-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GFHYISDTIWZUSU-QWRGUYRKSA-N 0.000 description 2
- LOOCQRRBKZTPKO-AVGNSLFASA-N Tyr-Glu-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 LOOCQRRBKZTPKO-AVGNSLFASA-N 0.000 description 2
- LUMQYLVYUIRHHU-YJRXYDGGSA-N Tyr-Ser-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LUMQYLVYUIRHHU-YJRXYDGGSA-N 0.000 description 2
- JFAWZADYPRMRCO-UBHSHLNASA-N Val-Ala-Phe Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JFAWZADYPRMRCO-UBHSHLNASA-N 0.000 description 2
- HZYOWMGWKKRMBZ-BYULHYEWSA-N Val-Asp-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N HZYOWMGWKKRMBZ-BYULHYEWSA-N 0.000 description 2
- BZWUSZGQOILYEU-STECZYCISA-N Val-Ile-Tyr Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 BZWUSZGQOILYEU-STECZYCISA-N 0.000 description 2
- ODUHAIXFXFACDY-SRVKXCTJSA-N Val-Val-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)C(C)C ODUHAIXFXFACDY-SRVKXCTJSA-N 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- UGXQOOQUZRUVSS-ZZXKWVIFSA-N [5-[3,5-dihydroxy-2-(1,3,4-trihydroxy-5-oxopentan-2-yl)oxyoxan-4-yl]oxy-3,4-dihydroxyoxolan-2-yl]methyl (e)-3-(4-hydroxyphenyl)prop-2-enoate Chemical compound OC1C(OC(CO)C(O)C(O)C=O)OCC(O)C1OC1C(O)C(O)C(COC(=O)\C=C\C=2C=CC(O)=CC=2)O1 UGXQOOQUZRUVSS-ZZXKWVIFSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 2
- 108010060035 arginylproline Proteins 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 108010004073 cysteinylcysteine Proteins 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 2
- YMAWOPBAYDPSLA-UHFFFAOYSA-N glycylglycine Chemical compound [NH3+]CC(=O)NCC([O-])=O YMAWOPBAYDPSLA-UHFFFAOYSA-N 0.000 description 2
- 108010015792 glycyllysine Proteins 0.000 description 2
- 108010040030 histidinoalanine Proteins 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 229940070765 laurate Drugs 0.000 description 2
- 108010009298 lysylglutamic acid Proteins 0.000 description 2
- 108010017391 lysylvaline Proteins 0.000 description 2
- 235000009973 maize Nutrition 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 2
- 108010015796 prolylisoleucine Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 108010073969 valyllysine Proteins 0.000 description 2
- HSTOKWSFWGCZMH-UHFFFAOYSA-N 3,3'-diaminobenzidine Chemical compound C1=C(N)C(N)=CC=C1C1=CC=C(N)C(N)=C1 HSTOKWSFWGCZMH-UHFFFAOYSA-N 0.000 description 1
- OVWSNDOUOHKMOM-UHFFFAOYSA-N 3-phenylprop-2-enoic acid;hydrochloride Chemical compound Cl.OC(=O)C=CC1=CC=CC=C1 OVWSNDOUOHKMOM-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 101000765308 Aspergillus niger N-(5'-phosphoribosyl)anthranilate isomerase Proteins 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 description 1
- 235000014698 Brassica juncea var multisecta Nutrition 0.000 description 1
- 240000002791 Brassica napus Species 0.000 description 1
- 235000006008 Brassica napus var napus Nutrition 0.000 description 1
- 235000006618 Brassica rapa subsp oleifera Nutrition 0.000 description 1
- 244000188595 Brassica sinapistrum Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 238000007399 DNA isolation Methods 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 244000000626 Daucus carota Species 0.000 description 1
- 235000002767 Daucus carota Nutrition 0.000 description 1
- LBQZVWQOPFFQJI-GGWOSOGESA-N Diferulic acid Natural products COC1=CC(\C=C\C(O)=O)=CC(C=2C(=C(OC)C=C(\C=C\C(O)=O)C=2)O)=C1O LBQZVWQOPFFQJI-GGWOSOGESA-N 0.000 description 1
- UESYBOXFJWJVSB-AVGNSLFASA-N Gln-Phe-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O UESYBOXFJWJVSB-AVGNSLFASA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 241000235649 Kluyveromyces Species 0.000 description 1
- 241000186610 Lactobacillus sp. Species 0.000 description 1
- 241000178948 Lactococcus sp. Species 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 description 1
- 241000235395 Mucor Species 0.000 description 1
- 241000233892 Neocallimastix Species 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- UNBFGVQVQGXXCK-KKUMJFAQSA-N Phe-Ser-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O UNBFGVQVQGXXCK-KKUMJFAQSA-N 0.000 description 1
- RGMLUHANLDVMPB-ULQDDVLXSA-N Phe-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N RGMLUHANLDVMPB-ULQDDVLXSA-N 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 235000021536 Sugar beet Nutrition 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- WBAJDGWKRIHOAC-GVXVVHGQSA-N Val-Lys-Gln Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O WBAJDGWKRIHOAC-GVXVVHGQSA-N 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 238000010805 cDNA synthesis kit Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 210000004534 cecum Anatomy 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000010931 ester hydrolysis Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000011121 hardwood Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229940059442 hemicellulase Drugs 0.000 description 1
- 108010002430 hemicellulase Proteins 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- VGKONPUVOVVNSU-UHFFFAOYSA-N naphthalen-1-yl acetate Chemical compound C1=CC=C2C(OC(=O)C)=CC=CC2=C1 VGKONPUVOVVNSU-UHFFFAOYSA-N 0.000 description 1
- WWZKQHOCKIZLMA-UHFFFAOYSA-M octanoate Chemical compound CCCCCCCC([O-])=O WWZKQHOCKIZLMA-UHFFFAOYSA-M 0.000 description 1
- 238000001139 pH measurement Methods 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 235000008476 powdered milk Nutrition 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 210000004767 rumen Anatomy 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000011122 softwood Substances 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Animal Husbandry (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Paper (AREA)
- Fodder In General (AREA)
Abstract
An enzyme, having phenolic acid esterase activity (preferably ferulic acid activity and/or coumaric acid esterase activity), which may be isolated from Piromyces spp. (preferably P.equi), is characterised by a pH optimum of greater than 6.5 and a temperature optimum greater than 45 degrees C. (as measured in a citric acid/disodium hydrogen orthophosphate buffer comprising 33 micromoles of FAXX (O-{5-O-[(E)-feruloyl]- alpha-L-arabinofuranosyl}-(1-3)-O-beta-D-xylopyranosyl-(1-4)-D-xylopyranose) as the substrate). DNA sequences encoding the enzyme, which may inserted into a vector and expressed in a transformed cell, are disclosed. The enzyme may in the form of a preparation having at least one further polysaccharide modifying and/or degrading enzyme, preferably a glycosyl hydrolase (especially xylanase, arabinase, alpha-L-arabinofuranosidase, endoglucanase, alpha-D-glucuronidase, pectinase, acetyl esterase, mannanase and acetyl xylan esterase). Such preparations may further comprise amylase, protease, alpha-galactosidase, phytase and/or lipase. The enzyme or enzyme preparation may be used in the preparation or release of phenolic acids from substrates comprising phenolic acid moieties, in the production of foodstuffs (including animal feed), in paper production, in the bioconversion of plant material or lignocellulose waste to sugars, and in the improvement of the digestibility of crop plants for livestock.
Description
Phenolic Acid Esterase and Use Thereof
The present invention relates to an enzyme with phenolic acid esterase activity, DNA molecule encoding said enzyme as well as a method for the production and use of said enzyme.
Background
Plant cell walls are divided into two sections, the primary and secondary cell wall. The primary cell wall is comprised of three major classes of polysaccharides: cellulose, hemicellulose and pectin. The secondary cell wall also contains polysaccharides as well as lignin. Hemicellulose, a general class of highly branched polysaccharides found in the plant cell wall, is bound to itself as well as to cellulose and lignin, and these bonds contribute to the stability and support of the plant structure.
Hemicelluloses based on a xylose backbone are designated as xylans. Xylan, which has been shown to exist in a wide variety of different plants including fruits, vegetables legumes, cereals, grasses, softwoods and hardwoods, is a linear j3-(l-4)-D-xylopyranose polymer which can be substituted with sugar residues, including a-L-arabinose, and a-Dglucuronic acid and/or the 4-0-methyl ether derivative of a-Dglucuronic acid. Many xylans are also esterified with phenolic acid residues, including coumaric acid and ferulic acid. These phenolic acid residues are present in an ester linkage to a-L-arabinofuranosyl xylan and can serve to protect xylan from xylan-degrading enzymes, so-called xylanases, as well they confer structural stability to the plant cell wall by forming covalent bonds with the lignin present therein.
In addition, ferulic acid has been shown to exist as a diferulic acid bridge between different xylan chains, imparting further structural support for plant cells (Linden,
J.C. et al., American Chemical Society Symposium Series, vol.
566 (1994), 452-467).
A number of microorganisms are known which are capable of hydrolysing phenolic acid esters and digesting plant cell walls through the enzymatic breakdown of plant cell wall polysaccharides. Some of these microorganisms possess enzyme(s) with phenolic acid esterase activity, i.e. coumaric acid esterase activity or ferulic acid esterase activity or a combination of these two activities.
For example, Borneman, W.S. et al (Applied and Environmental
Microbiology, vol. 58 (1992), 3762-3766) describe two ferulic acid esterases (FAE), designated FAE-I and FAE-II respectively, isolated from the anaerobic fungus
Neocallimastix strain MC-2. FAE-II was reported to be specific for the substrate (o-(5-0-[(E)-feruloyl]-a-L- arabinofuranosyl)- (1-3) -o-P-D-xylopyranosyl- (1-4) -D- xylopyranose (FAXX), whereas FAE-I was reported to have both a
FAXX degrading activity as well as a (0-(5-0-[(E)-p- coumaroyl]-a-L-arabinofuranosyl}-(1-3)-O-ss-D-xylopyranosyl-(1- 4)-D-xylopyranose (PAXX) degrading activity, the maximum ratio of metabolism of FAXX:PAXX being 3:1. The pH optima of these two enzymes were shown to be 6.2 and 7.0 respectively when using FAXX as a substrate.
GB 2 301 103 discloses an FAE obtained from Aspergillus niger as well as the gene encoding said enzyme. Said enzyme has a pH optimum of about 5 and a temperature optimum of from about 50 to 600C when methyl ferulate is used as a substrate.
Other purified enzymes with ferulic acid esterase activity are known (for example, see McCrae, S.I. et al., Enzyme Microb.
Technol., vol. 16 (1994), 826-834; Faulds, C.B. and
Williamson, G., Microbiology, vol. 140 (1994), 779-787;
Castanares, A. et al., Enzyme Microb. Technol., vol. 14 (1992), 875; and Kroon, P.A. et al., Biotechnol. Appl.
Biochem. vol 23 (1996), 255-262) which have pH optima ranging from about 5.0 to 6.0 and temperature optima from 30 to 600C.
Enzymes with phenolic acid esterase activity can be used in a number of industrial, agricultural and health applications which can be carried out at pH values about or above 6.5 and/or at temperatures above 450C.
Summary of the invention
It is an object of the present invention to provide enzyme with good phenolic acid esterase activity.
In addition, it is an object of the present invention to provide a source of an enzyme with phenolic acid esterase activity which is available in relatively large amounts.
Furthermore, it is an object to provide a method for the production of an enzyme with phenolic acid esterase activity.
A further object is to provide uses of an enzyme phenolic acid esterase activity for the preparation of food and feed, for the processing of paper and pulp as well as for the bioconversion of ligno-cellulose wastes, for example.
Other objects of the present invention will become apparent from the following detailed description.
Subject matter of the present invention is an enzyme with phenolic acid esterase activity, characterized in that said enzyme has a pH optimum greater than pH 6.5, preferably about 7.0, and a temperature optimum greater than 45"C, preferably greater than 500C, and more preferably about 550C, when measured in a citric acid/disodium hydrogen orthophosphate buffer containing 33 ssM FAXX as a substrate. Preferably, said enzyme has ferulic acid esterase activity and coumaric acid esterase activity.
Subject matter of the present invention is also an enzyme with phenolic acid esterase activity obtainable from Piromyces Sp., for example Piromyces equi, and more preferably from the
Piromyces equi strain deposited under the Budapest Treaty at the International Mycological Institute (IMI), Bakeham Lane,
Egham, Surrey, UK under the Accession Number 375061.
Preferably, the enzyme of the present invention comprises the amino acid sequence given in SEQ ID NO:1 or a functional derivative thereof. A functional derivative of the enzyme of the present invention is defined as an enzyme having one or more N-terminal, C-terminal or internal substitution(s), insertion(s) and/or deletion(s) in the amino acid sequence given in SEQ ID NO:1 which maintains a pH optimum greater than 6.5, preferably about 7.0, and a temperature optimum greater than 450C, preferably greater than 500C, and more preferably about 550C, when measured in a citric acid/disodium hydrogen orthophosphate buffer containing 33 AM FAXX as a substrate.
More preferably, the enzyme of the present invention comprises the amino acid sequence given in SEQ ID NO:3 or a functional derivative thereof.
In addition, the enzyme of the present invention is preferably encoded by the DNA sequence given in SEQ ID NO:1 or a functional derivative thereof. More preferably, the enzyme of the present invention is encoded by the DNA sequence given in
SEQ ID NO:3 or a functional derivative thereof.
The present invention relates to a phenolic acid esterase with one or more of the above properties.
Further subject matter of the present invention is a DNA molecule encoding an enzyme with phenolic acid esterase activity, characterized in that said DNA molecule comprises a
DNA sequence as given in SEQ ID NO:1 or a functional derivative or homologue thereof. A functional derivative of the DNA sequence given in SEQ ID NO:1 is defined as a DNA sequence having one or more 5'-, 3'- or internal substitution(s), insertion(s) and/or deletion(s) in the DNA sequence given in SEQ ID NO:1 which maintains its capability to encode an enzyme with phenolic acid esterase activity which has a pH optimum greater than 6.5, preferably about 7.0, and a temperature optimum greater than 450C, preferably greater than 500C, and more preferably about 550C, when measured in a citric acid/disodium hydrogen orthophosphate buffer comprising 33 yM FAXX as a substrate. A functional homologue of the DNA sequence of the present invention is defined as a DNA sequence with preferably 75 homology, more preferably 85% homology and most preferably 95% homology to the DNA sequence given in SEQ
ID NO:1 or SEQ ID NO:3. More preferably, a DNA molecule encoding an enzyme according to the present invention comprises a DNA sequence as given in SEQ ID NO:3 or a functional derivative or homologue thereof.
In a preferred embodiment, DNA molecules of the present invention comprise vector sequence capable of replicating said
DNA molecules and/or expressing said enzyme in a procaryotic or eucaryotic host.
Further subject matter of the present invention is a transformed procaryotic cell or eucaryotic cell comprising one or more DNA molecules of the present invention. Preferably said cells are selected from the group comprising E. coli,
Bacillus sp., such as Bacillus subtilis, Lactobacillus sp., and Lactococcus sp., Aspergillus, Trichoderma, Penicillium,
Mucor, Kluyveromyces and Saccharomyces, such as Saccharomyces cerevisiae.
The enzyme of the present invention may be expressed in transgenic plants such as maize, soybean and canola/rapeseed.
or in root storage organs of plants, such as potato, carrot and sugar beet.
The introduction of an esterase of the present invention expressed and/or secreted at the appropriate stage, for example, at harvest, has the advantage that the risk of weakening the transgenic plant or storage root organ structure during growth can be reduced.
The methodology for the production of transformed procaryotic and eucaryotic cells is known in the art. Transgenic fungus, such as Aspergillus, tranformed yeast, such as Saccharomyces, and transgenic plants are also known inthe art and can be produced by the methods taught and discussed in GB 2 301 103,
EP 479 359 and EP 449 375.
Subject matter of the present invention is also a method for the production of an enzyme or enzyme preparation having phenolic acid esterase activity, characterized in that said enzyme is isolated from a naturally occurring organism or transformed cell or organism capable of expressing the enzyme according to the invention. Enzyme preparations including, for example, partially purified preparations obtainable as a cell or organism extract are also subject matter of the present invention.
The enzyme preparation of the present invention can comprise one or more further polysaccharide modifying and/or degrading enzymes. Said polysaccharide modifying and/or degrading enzyme(s) is (are) preferably selected from the group comprising xylanase, arabinanase, o'-L-arabinofuranosidase, endoglucanase, a-D-glucuronidase, pectinase, acetyl esterase, mannanase, acetyl xylan esterase and other glycosyl hydrolases.
In addition, the enzyme preparation of the present invention can also include one or more further enzymes selected from the group comprising amylase, protease, cc-galactosidase, phytase and lipase.
Use of the enzyme and/or enzyme preparation according to the invention include the use in a process for releasing phenolic acids from a substrate comprising phenolic acid moieties.
Said enzyme and/or enzyme preparation according to the invention can equally find use in the production of animal feed by improving the digestibility of plant material, especially forage in which the plant cell walls have a high phenolic acid content. Furthermore, the enzyme and/or the enzyme preparation according to the invention can be used in or with crop plants including but not limited to maize, wheat, grasses and alfalfa, to improve the digestability for livestock by pre-modifying the cell wall content. Said enzyme and/or enzyme preparation according to the invention can also find used in the preparation of food for human consumption.
Further subject matter of the present invention is also a feed additive comprising an enzyme or enzyme preparation having phenolic acid esterase activity according to the invention and a feed comprising said feed additive.
The enzyme and/or enzyme preparation according to the invention can also find use in the paper and pulp industry, for example, in helping remove lignin from cellulose pulps.
Additionally, used in combination with xylan degrading enzymes, the enzyme and/or enzyme preparation according to the invention can contribute to a reduction in the amount of chlorine required for bleaching by increasing the solubility and extractability of lignin from pulp.
Furthermore, when combined with xylanases and/or cellulases, the enzyme and/or enzyme preparation according to the invention can be used for the bioconversion of plant material or ligno-cellulose wastes to sugars, for example, for chemical or fuel production, and/or in the production of phenolic acids.
Brief description of the drawings
Figure 1: pH profile of the phenolic acid esterase of the
present invention measured using FAXX as a
substrate.
Figure 2: Temperature profile of the phenolic acid esterase of
the present invention measured using FAXX as a
substrate.
Detailed description of the invention
The following Examples are intended to more closely illustrate the present invention without limiting the subject matter of the invention to said Examples.
Example 1
Piromyces equi isolated from horse cecum (Orpin, C.G., J. Gen.
Microbiol., vol. 123 (1981), 287-296) and as described by E.A.
Munn in Anaerobic Fungi, Biology, Ecology and Function, D.O.
Mountfort and C.G. Orpin Eds., Marcel Dekker, Inc., New York, 1994, 47-1OS, and deposited under the Budapest Treaty at the
International Mycological Institute (IMI) under the Accession
Number 375061 was cultured under anaerobic conditions at a temperature of 390C in a rumen fluid-containing medium (Kemp,P., Lander, D.J. and Orpin, C.G., J. Gen. Microbiol., vol. 130 (1984), 27-37) with 0.10% soluble xylan and 0.5W Sigmacell (Sigma Chemical Co., Poole, Dorset, England) as carbon sources. Total RNA was extracted from fungus grown under the above conditions, poly(A)+ RNA was selected by oligo(dT) chromatography, and double-stranded cDNA was synthesized from the selected RNA, cloned into XZAPII using a
ZAP-cDNA synthesis kit and packaged in vitro according to the instructions of the manufacturer (Stratagene, La Jolla,
California, USA) (Xue, G-P. et al., J Gen. Microbiol., vol.
138 (1992), 1413-1420 and Ali, B.R.S. et al., FEMS Microbiol.
Lett., vol. 125 (1995), 15-22). Recombinant phage were grown by plating on lawns of E. coli XL1-Blue in soft agar overlays and screened using an antibody raised against a fungal cellulase/hemicellulase complex purified according to
Ali,B.R.S. et al., FEMS Microbiol. Lett., vol. 125 (1995), 1522). Antibody screening of phage plaques with rabbit anticomplex antibody as the primary antibody was carried out essentially as described in the instruction manual provided with the picoBlueN immunoscreening kit (Stratagene), with the following modifications: isopropyl- -D-thiogalactopyranoside (IPTG: 0.33 mM) was added directly to the soft agar overlays containing recombinant XZAPII and host bacteria (E. coli XL1
Blue); plaques were lifted onto Hybond-C filters (Amersham); blocking solution contained dried milk powder (4W w/v) in place of BSA; anti-rabbit IgG (whole molecule) conjugated to horseradish peroxidase (Sigma Chemical Co.) was used as secondary antibody; colour development solution comprised 3,3'-diaminobenzidine (0.5 mg/ml) in 50 mM Tris-HCl buffer, pH 7.4, containing hydrogen peroxide (0.5 yl/ml. Esterase production was verified by showing that a clone selected by antibody screening synthesized an enzyme which hydrolysed [4methylumbelliferoyl (p-trimethylammonium cinnamate chloride)] according to Dalrymple, B.P. et al., FEMS Microbiol. Lett., vol 143 (1996), 115-120.
General molecular biological techniques including DNA isolation, restriction endonuclease digestion, ligation, transformation as well as DNA sequencing of the esterase gene were performed in accordance with Sambrook et al., Molecular
Cloning: A Laboratory Manual, 2nd. Ed. (1989), Cold Spring
Harbor, New York.
Nucleotide sequencing of the the gene encoding the enzyme having phenolic acid esterase activity of the present invention was performed and the results are given in SEQ ID
NO:3. The open reading frame comprises 1608 nucleotides, encoding a protein of 536 amino acids with a predicted molecular weight of 55,540 daltons.
Example 2
Measurement of pH optimum
A truncated enzyme encoded by SEQ ID NO. 1 was generated in a
PCR reaction (20 cycles of 30 seconds at 940C, 45 seconds at 50 C, and 1 minute at 720C) in a buffer comprising 50 mM Tris
HCl, pH 9.0, 50 mM NaCl, 10 mM MgC12, 200 gM dNTPs, 50 picomoles of the primers 5'-end: 5' -CGCGGATCCAACAGCGGTCCAACTGTTG-3' 3'-end: 5' -GCGAATTCTTATCTTATGGGAGAGAG-3', and 250 ng template DNA and expressed in E.coli BL21 (DE3) (Novagen, Inc., Wisconsin, USA) using the vector pET32a (Novagen, Inc.).
The enzyme was purified from freshly prepared cell-free extracts by binding to Talon resin (Clontech Laboratories
Inc., California, USA) and cleaved from the metal affinity resin using restriction grade Thrombin (Sigma) in accordance to the guidelines provided by Novagen, Inc., USA, for use with pET vectors. The enzyme was further purified as follows: a 1 ml MonoQ column (Pharmacia) was equilibrated with 10 mM Tris, pH 8.0, and fresh enzyme was applied. The enzyme was eluted at 1.0 ml/min with a sodium chloride gradient (0 to 0.5 M NaCl in 10 mM Tris, pH 8.0). Fractions of 1.0 ml were collected.
The enzyme was assayed in McIlvaine's buffer (citric acid/ disodium hydrogen orthophosphate, see Data for Biochemical
Research, 3rd Edition, Dawson, Elliot, Elliot, Jones, Oxford
Science Publications, Oxford University Press, 1987) for pH values ranging from 3 to 7 or a buffer comprising potassium chloride/ boric acid for pH values ranging from 8 to 9. The assay was carried out at 370C with a final FAXX concentration of 33 yM. Ferulic acid release from FAXX was monitored continuously for 3 min at 335 nm according to (Faulds, C.B.
and Williamson, G., Microbiology, vol. 140 (1994), 779-787).
The results of the assay are given in Figure 1. As can be deduced from Figure 1, the enzyme of the invention exhibited 50% activity at about pH 5.5 and about 8.5
In order to determine the temperature optimum of the enzyme according to the invention using FAXX as a substrate, FAXX was employed at a concentration of 33 yM and the assay was performed at pH 6.0 in 100 mM MOPS buffer. The temperature of incubation was changed from 200C to 700C using a thermostatically controlled spectrophotometer. The release of ferulic acid from FAXX was measured at 335 nm as described above. The results are presented in Figure 2.
Kinetics
The Km and Vmax of the enzyme of the present invention were determined using FAXX and Ara2F (o-[2-o(trans-feru1oyl)-- arabinofuranosyl]- (1-5) -L-arabinofuranose) as substrates. FAXX was employed at concentrations varying from 3.72 yM to 49.18 yM and Ara2F was used at concentrations ranging from 4.46 /M to 122.92 yM. The assay was performed at 370C and pH 6.0 in 100 mM MOPS ((3-[N-morpholino]propanesulfonic acid)) buffer with 90 ng enzyme. For both substrates, the release of ferulic acid was measured at 335 nM as described above.
Based on results of the above experiments, it was determined that the enzyme of the present invention has the following kinetic constants: substrate Km Vmax
FAXX 3.0+0.3 yM 35.6+0.9 ymol/min/mg Ara2F 234+27 ZM 19.6+1.7 mol/min/mg.
Hence, the enzyme of the present invention has a Km of about 3.0 and a Vrnax of about 35 when measured under the above conditions using FAXX as a substrate.
The specific activity of the enzyme of the present invention was determined for methyl ferulate, methyl coumarate and methyl p-coumarate in an assay at 370C comprising 100 mM MOPS buffer (with 0.02% azide), pH 6.0., 44 ng enzyme and 1 mM of the above substrates. After 15 minutes incubation time, the reaction was terminated by boiling and the free acid liberated was measured using reverse phase HPLC (Kroon, P.A. and
Williamson, G., Biotechnol. Appl. Biochem., vol. 23 (1996), 263-267). The results of the above experiment are shown below.
In addition, the specific activity of the enzyme of the present invention was determined for p-nitrophenyl acetate, anaphthyl acetate, a-naphthyl butyrate, a-naphthyl caproate anaphthyl caprylate and a-naphthyl laurate according to the methods described in Ferreira, L.M.A. et al.(Biochem. J., vol.
294 (1993), 349-355). The results of the above assay are shown below.
substrate specific activity (U*/mg) p-nitrophenyl acetate 204.3 a-naphthyl acetate 121 a-naphthyl butyrate 220 a-naphthyl caproate 256 a-naphthyl caprylate 54 a-naphthyl laurate 6 methyl ferulate 10.6 methyl coumarate 10.5 methyl p-coumarate 2.7 *1 U is defined as the amount of enzyme which gives 1 Zmol/min of ester hydrolysis.
SEQUENCE LISTING (1) GENERAL INFORMATION:
(i) APPLICANT:
(A) NAME: Biotechnology and Biological Sciences Research Council
(B) STREET:Polaris House, North Star Avenue
(C) CITY: Swindon
(D) STATE:
(E) COUNTRY: United Kingdom
(F) POSTAL CODE (ZIP): SN2 1UH
(ii) TITLE OF INVENTION: Phenolic Acid Esterase and Use Thereof
(iii) NUMBER OF SEQUENCES: 4
(iv) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk
(B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: PatentIn Release #1.0, Version #1.30 (EPO) (2) INFORMATION FOR SEQ ID NO: 1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 825 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA (genomic)
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Piromyces equi
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..822
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:
AAC AGC GGT CCA ACT GTT GAA TAC TCT ACT GAT GTT GAC TGT TCC GGT 48
Asn Ser Gly Pro Thr Val Glu Tyr Ser Thr Asp Val Asp Cys Ser Gly
1 5 10 15
AAG ACC CTT AAG AGT AAC ACC AAC CTT AAC ATC AAT GGT CGT AAG GTT 96
Lys Thr Leu Lys Ser Asn Thr Asn Leu Asn Ile Asn Gly Arg Lys Val
20 25 30
ATT GTA AAA TTC CCA AGC GGC TTC ACT GGT GAC AAG GCT GCT CCA CTT 144
Ile Val Lys Phe Pro Ser Gly Phe Thr Gly Asp Lys Ala Ala Pro Leu
35 40 45
CTT ATT AAC TAC CAT CcA ATT ATG GGT AGT GCT TCT CAA TGG GAA AGT 192
Leu Ile Asn Tyr His Pro Ile Met Gly Ser Ala Ser Gln Trp Glu Ser
50 55 60
GGT TCT CAA ACT GCT AAG GCT GCT TTA AAT GAT GGT GCC ATC GTT GCT 240
Gly Ser Gln Thr Ala Lys Ala Ala Leu Asn Asp Gly Ala Ile Val Ala
65 70 75 80
TTC ATG GAT GGT GCT CAA GGT CCA ATG GGA CAA GCT TGG AAC GTT GGT 288
Phe Met Asp Gly Ala Gln Gly Pro Met Gly Gln Ala Trp Asn Val Gly
85 90 95
CCA TGT TGT ACT GAT GCT GAT GAT GTT CAA TTC ACT CGT AAC TTC ATT 336
Pro Cys Cys Thr Asp Ala Asp Asp Val Gln Phe Thr Arg Asn Phe Ile
100 105 110
AAG GAA ATC ACT AGT AAG GCT TGT GTT GAT CCA AAG CGT ATC TAT GCT 384
Lys Glu Ile Thr Ser Lys Ala Cys Val Asp Pro Lys Arg Ile Tyr Ala
115 120 125
GCT GGT TTC TCT ATG GGT GGT GGT ATG TCT AAC TAT GCT GGT TGT CAA 432
Ala Gly Phe Ser Met Gly Gly Gly Met Ser Asn Tyr Ala Gly Cys Gln
130 135 140
CTT GCT GAT GTT ATT GCT GCT GCT GCT CCA TCA GCC TTT GAT CTT GCC 480
Leu Ala Asp Val Ile Ala Ala Ala Ala Pro Ser Ala Phe Asp Leu Ala 145 150 155 160
AAG GAA ATT GTT GAT GGT GGT AAA TGT AAA CCA GCT CGT CCA TTC CCA 528
Lys Glu Ile Val Asp Gly Gly Lys Cys Lys Pro Ala Arg Pro Phe Pro
165 170 175
ATC CTT AAC TTC CGT GGT ACT CAA GAT AAC GTT GTT ATG TAC AAC GGT 576
Ile Leu Asn Phe Arg Gly Thr Gln Asp Asn Val Val Met Tyr Asn Gly
180 185 190
GGT CTT TCT CAA GTT GTT CAA GGT AAG CCA ATT ACT TTC ATG GGT GCC 624
Gly Leu Ser Gln Val Val Gln Gly Lys Pro Ile Thr Phe Met Gly Ala
195 200 205
AAG AAC AAC TTC AAG GAA TGG GCT AAG ATG AAC GGA TGT ACT GGT GAA 672
Lys Asn Asn Phe Lys Glu Trp Ala Lys Met Asn Gly Cys Thr Gly Glu
210 215 220
CCA AAA CAA AAC ACT CCA GGT AAC AAC TGT GAA ATG TAC GAA AAC TGT 720
Pro Lys Gln Asn Thr Pro Gly Asn Asn Cys Glu Met Tyr Glu Asn Cys 225 230 235 240
AAG GGT GGT GTT AAG GTT GGT CTT TGC ACT ATC AAC GGT GGT GGT CAC 768
Lys Gly Gly Val Lys Val Gly Leu Cys Thr Ile Asn Gly Gly Gly His
245 250 255
GCT GAA GGT GAC GGT AAA ATG GGT TGG GAC TTT GTT AAA CAA TTC TCT 816
Ala Glu Gly Asp Gly Lys Met Gly Trp Asp Phe Val Lys Gln Phe Ser
260 265 270 CTC,CCA 825 Leu Pro (2) INFORMATION FOR SEQ ID NO: 2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 274 amino acids
(B) TYPE: amino acid
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:
Asn Ser Gly Pro Thr Val Glu Tyr Ser Thr Asp Val Asp Cys Ser Gly
1 5 10 15
Lys Thr Leu Lys Ser Asn Thr Asn Leu Asn Ile Asn Gly Arg Lys Val
20 25 30
Ile Val Lys Phe Pro Ser Gly Phe Thr Gly Asp Lys Ala Ala Pro Leu
35 40 45
Leu Ile Asn Tyr His Pro Ile Met Gly Ser Ala Ser Gln Trp Glu Ser
50 55 60
Gly Ser Gln Thr Ala Lys Ala Ala Leu Asn Asp Gly Ala Ile Val Ala
65 70 75 80
Phe Met Asp Gly Ala Gln Gly Pro Met Gly Gln Ala Trp Asn Val Gly
85 90 95
Pro Cys Cys Thr Asp Ala Asp Asp Val Gln Phe Thr Arg Asn Phe Ile
100 105 110
Lys Glu Ile Thr Ser Lys Ala Cys Val Asp Pro Lys Arg Ile Tyr Ala
115 120 125
Ala Gly Phe Ser Met Gly Gly Gly Met Ser Asn Tyr Ala Gly Cys Gln
130 135 140
Leu Ala Asp Val Ile Ala Ala Ala Ala Pro Ser Ala Phe Asp Leu Ala 145 150 155 160
Lys Glu Ile Val Asp Gly Gly Lys Cys Lys Pro Ala Arg Pro Phe Pro
165 170 175
Ile Leu Asn Phe Arg Gly Thr Gln Asp Asn Val Val Met Tyr Asn Gly
180 185 190
Gly Leu Ser Gln Val Val Gln Gly Lys Pro Ile Thr Phe Met Gly Ala
195 200 205
Lys Asn Asn Phe Lys Glu Trp Ala Lys Met Asn Gly Cys Thr Gly Glu
210 215 220
Pro Lys Gln Asn Thr Pro Gly Asn Asn Cys Glu Met Tyr Glu Asn Cys 225 230 235 240
Lys Gly Gly Val Lys Val Gly Leu Cys Thr Ile Asn Gly Gly Gly His
245 250 255
Ala Glu Gly Asp Gly Lys Met Gly Trp Asp Phe Val Lys Gln Phe Ser
260 265 270
Leu Pro (2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1611 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA (genomic)
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Piromyces equi
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..1608
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:
ATG AAG ACT AGC ATT GTA TTA TCT ATC GTT GCT TTA TTT TTA ACA TCC 48
Met Lys Thr Ser Ile Val Leu Ser Ile Val Ala Leu Phe Leu Thr Ser 275 280 285 290
AAA GCT TCT GCT GAT TGT TGG TCA GAA AGA TTA GGT TGG CCA TGC TGT 96
Lys Ala Ser Ala Asp Cys Trp Ser Glu Arg Leu Gly Trp Pro Cys Cys
295 300 305
AGT GAC AGC AAT GCC GAA GTA ATC TAC GTC GAT GAC GAT GGT GAT TGG 144
Ser Asp Ser Asn Ala Glu Val Ile Tyr Val Asp Asp Asp Gly Asp Trp
310 315 320
GGT GTT GAA AAT AAT GAC TGG TGT GGT ATC CAA AAG GAA GAA GAA AAC 192
Gly Val Glu Asn Asn Asp Trp Cys Gly Ile Gln Lys Glu Glu Glu Asn
325 330 335
AAT AAC TCA TGG GAT ATG GGT GAT TGG AAC CAA GGT GGT AAC CAA GGT 240
Asn Asn Ser Trp Asp Met Gly Asp Trp Asn Gln Gly Gly Asn Gln Gly
340 345 350
GGC GGT ATG CCA TGG GGC GAC TTT GGC GGT AAC CAA GGT GGT GGT ATG 288
Gly Gly Met Pro Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly Gly Met 355 , 360 365 370
CAA TGG GGT GAC TTC GGT GGT AAC CAA GGT GGC GGT ATG CCA TGG GGC 336
Gln Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly
375 380 385
GAC TTC GGT GGT AAC CAA GGT GGC GGT ATG CCA TGG GGT GAC TTT GGC 384
Asp Phe Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe Gly
390 395 400
GGT AAC CAA GGT GGT AAC CAA GGC GGT GGT ATG CCA TGG GGC GAC TTT 432
Gly Asn Gln Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe
405 410 415
GGA GGA AAC CAA GGA GGT AAC CAA GGT GGC GGT ATG CCA TGG GGT GAT 480
Gly Gly Asn Gln Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp
420 425 430
TTC GGA GGT AAC CAA GGT GGT GGT ATG CAA TGG GGC GAC TTT GGA GGA 528
Phe Gly Gly Asn Gln Gly Gly Gly Met Gln Trp Gly Asp Phe Gly Gly 435 440 445 450
AAC CAA GGA GGT AAC CAA GGT GGC GGT ATG CCA TGG GGT GAT TTC GGA 576
Asn Gln Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe Gly
455 460 465
GGT AAC CAA GGT GGT GGT ATG CAA TGG GGC GAC TTT GGA GGA AAC CAA 624
Gly Asn Gln Gly Gly Gly Met Gln Trp Gly Asp Phe Gly Gly Asn Gln
470 475 480
GGA GGT AAC CAA GGT GGC GGT ATG CCA TGG GGT GAC TTC GGA GGT AAC 672
Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe Gly Gly Asn
485 490 495
CAA GGT GGT GGT ATG CAA TGG GGC GAT TTC GGA GGT AAT CAA GGT GGT 720
Gln Gly Gly Gly Met Gln Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly
500 505 510
GGT ATG CAA TGG GGC GAC TTC GGC GGT AAC CAA GGA GGT AAC CAA GAT 768
Gly Met Gln Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly Asn Gln Asp 515 520 525 530
TGG GGT AAC CAA GGT GGT AAC AGC GGT CCA ACT GTT GAA TAC TCT ACT 816
Trp Gly Asn Gln Gly Gly Asn Ser Gly Pro Thr Val Glu Tyr Ser Thr
535 540 545
GAT GTT GAC TGT TCC GGT AAG ACC CTT AAG AGT AAC ACC AAC CTT AAC 864
Asp Val Asp Cys Ser Gly Lys Thr Leu Lys Ser Asn Thr Asn Leu Asn
550 555 560
ATC AAT GGT CGT AAG GTT ATT GTA AAA TTC CCA AGC GGC TTC ACT GGT 912
Ile Asn Gly Arg Lys Val Ile Val Lys Phe Pro Ser Gly Phe Thr Gly
565 570 575
GAC AAG GCT GCT CCA CTT CTT ATT AAC TAC CAT CCA ATT ATG GGT AGT 960
Asp Lys Ala Ala Pro Leu Leu Ile Asn Tyr His Pro Ile Met Gly Ser
580 585 590
GCT TCT CAA TGG GAA AGT GGT TCT CAA ACT GCT AAG GCT GCT TTA AAT 1008
Ala Ser Gln Trp Glu Ser Gly Ser Gln Thr Ala Lys Ala Ala Leu Asn 595 600 605 610
GAT GGT GCC ATC GTT GCT TTC ATG GAT GGT GCT CAA GGT CCA ATG GGA 1056
Asp Gly Ala Ile Val Ala Phe Met Asp Gly Ala Gln Gly Pro Met Gly
615 620 625
CAA GCT TGG AAC GTT GGT CCA TGT TGT ACT GAT GCT GAT GAT GTT CAA 1104
Gln Ala Trp Asn Val Gly Pro Cys Cys Thr Asp Ala Asp Asp Val Gln
630 635 640
TTC ACT CGT AAC TTC ATT AAG GAA ATC ACT AGT AAG GCT TGT GTT GAT 1152
Phe Thr Arg Asn Phe Ile Lys Glu Ile Thr Ser Lys Ala Cys Val Asp
645 650 655
CCA AAG CGT ATC TAT GCT GCT GGT TTC TCT ATG GGT GGT GGT ATG TCT 1200
Pro Lys Arg Ile Tyr Ala Ala Gly Phe Ser Met Gly Gly Gly Met Ser
660 665 670
AAC TAT GCT GGT TGT CAA CTT GCT GAT GTT ATT GCT GCT GCT GCT CCA 1248
Asn Tyr Ala Gly Cys Gln Leu Ala Asp Val Ile Ala Ala Ala Ala Pro 675 680 685 690
TCA GCC TTT GAT CTT GCC AAG GAA ATT GTT GAT GGT GGT AAA TGT AAA 1296
Ser Ala Phe Asp Leu Ala Lys Glu Ile Val Asp Gly Gly Lys Cys Lys
695 700 705
CCA GCT CGT CCA TTC CCA ATC CTT AAC TTC CGT GGT ACT CAA GAT AAC 1344
Pro Ala Arg Pro Phe Pro Ile Leu Asn Phe Arg Gly Thr Gln Asp Asn
710 715 720
GTT GTT ATG TAC AAC GGT GGT CTT TCT CAA GTT GTT CAA GGT AAG CCA 1392
Val Val Met Tyr Asn Gly Gly Leu Ser Gln Val Val Gln Gly Lys Pro
725 730 735
ATT ACT TTC ATG GGT GCC AAG AAC AAC TTC AAG GAA TGG GCT AAG ATG 1440
Ile Thr Phe Met Gly Ala Lys Asn Asn Phe Lys Glu Trp Ala Lys Met
740 745 750
AAC GGA TGT ACT GGT GAA CCA AAA CAA AAC ACT CCA GGT AAC AAC TGT 1488
Asn Gly Cys Thr Gly Glu Pro Lys Gln Asn Thr Pro Gly Asn Asn Cys 755 760 765 770
GAA ATG TAC GAA AAC TGT AAG GGT GGT GTT AAG GTT GGT CTT TGC ACT 1536
Glu Met Tyr Glu Asn Cys Lys Gly Gly Val Lys Val Gly Leu Cys Thr
775 780 785
ATC AAC GGT GGT GGT CAC GCT GAA GGT GAC GGT AAA ATG GGT TGG GAC 1584
Ile Asn Gly Gly Gly His Ala Glu Gly Asp Gly Lys Met Gly Trp Asp
790 795 800
TTT GTT AAA CAA TTC TCT CTC CCA TAA 1611
Phe,Val Lys Gln Phe Ser Leu Pro
805 810 (2) INFORMATION FOR SEQ ID NO: 4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 536 amino acids
(B) TYPE: amino acid
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:
Met Lys Thr Ser Ile Val Leu Ser Ile Val Ala Leu Phe Leu Thr Ser
1 5 10 15
Lys Ala Ser Ala Asp Cys Trp Ser Glu Arg Leu Gly Trp Pro Cys Cys
20 25 30
Ser Asp Ser Asn Ala Glu Val Ile Tyr Val Asp Asp Asp Gly Asp Trp
35 40 45
Gly Val Glu Asn Asn Asp Trp Cys Gly Ile Gln Lys Glu Glu Glu Asn
50 55 60
Asn Asn Ser Trp Asp Met Gly Asp Trp Asn Gln Gly Gly Asn Gln Gly
65 70 75 80
Gly Gly Met Pro Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly Gly Met
85 90 95
Gln Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly
100 105 110
Asp Phe Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe Gly
115 120 125
Gly Asn Gln Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe
130 135 140
Gly Gly Asn Gln Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp 145 150 155 160
Phe Gly Gly Asn Gln Gly Gly Gly Met Gln Trp Gly Asp Phe Gly Gly
165 170 175
Asn Gln Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe Gly
180 185 190
Gly Asn Gln Gly Gly Gly Met Gln Trp Gly Asp Phe Gly Gly Asn Gln
195 200 205
Gly Gly Asn Gln Gly Gly Gly Met Pro Trp Gly Asp Phe Gly Gly Asn
210 215 220
Gln Gly Gly Gly Met Gln Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly 225 230 235 240
Gly Met Gln Trp Gly Asp Phe Gly Gly Asn Gln Gly Gly Asn Gln Asp
245 250 255
Trp Gly Asn Gln Gly Gly Asn Ser Gly Pro Thr Val Glu Tyr Ser Thr
260 265 270
Asp Val Asp Cys Ser Gly Lys Thr Leu Lys Ser Asn Thr Asn Leu Asn
275 280 285
Ile Asn Gly Arg Lys Val Ile Val Lys Phe Pro Ser Gly Phe Thr Gly
290 295 300
Asp Lys Ala Ala Pro Leu Leu Ile Asn Tyr His Pro Ile Met Gly Ser 305 310 315 320
Ala Ser Gln Trp Glu Ser Gly Ser Gln Thr Ala Lys Ala Ala Leu Asn
325 330 335
Asp Gly Ala Ile Val Ala Phe Met Asp Gly Ala Gln Gly Pro Met Gly
340 345 350
Gln Ala Trp Asn Val Gly Pro Cys Cys Thr Asp Ala Asp Asp Val Gln
355 360 365
Phe Thr Arg Asn Phe Ile Lys Glu Ile Thr Ser Lys Ala Cys Val Asp
370 375 380
Pro Lys Arg Ile Tyr Ala Ala Gly Phe Ser Met Gly Gly Gly Met Ser 385 390 395 400
Asn Tyr Ala Gly Cys Gln Leu Ala Asp Val Ile Ala Ala Ala Ala Pro
405 410 415
Ser Ala Phe Asp Leu Ala Lys Glu Ile Val Asp Gly Gly Lys Cys Lys
420 425 430
Pro Ala Arg Pro Phe Pro Ile Leu Asn Phe Arg Gly Thr Gln Asp Asn
435 440 445
Val Val Met Tyr Asn Gly Gly Leu Ser Gln Val Val Gln Gly Lys Pro
450 455 460
Ile Thr Phe Met Gly Ala Lys Asn Asn Phe Lys Glu Trp Ala Lys Met 465 470 475 480
Asn Gly Cys Thr Gly Glu Pro Lys Gln Asn Thr Pro Gly Asn Asn Cys
485 490 495
Glu Met Tyr Glu Asn Cys Lys Gly Gly Val Lys Val Gly Leu Cys Thr
500 505 510
Ile Asn Gly Gly Gly His Ala Glu Gly Asp Gly Lys Met Gly Trp Asp
515 520 525
Phe Val Lys Gln Phe Ser Leu Pro
530 535
Claims (23)
- Claims: 1. Enzyme with phenolic acid esterase activity, characterized in that said enzyme has a pH optimum greater than pH 6.5 and a temperature optimum greater than 45"C when measured in a citric acid/disodium hydrogen orthophosphate buffer containing 33 yM FAXX as a substrate.
- 2. Enzyme according to claim 1, characterized in that said enzyme has ferulic acid esterase activity and/or coumaric acid esterase activity.
- 3. Enzyme according to claim 1 or 2, characterized in that said enzyme has a a pH optimum of about pH 7.0 and/or a temperature optimum of about 550C.
- 4. Enzyme according to any of calims 1 to 3, characterized in that said enzyme is obtainable from Piromyces Sp., preferably Piromyces equi deposited at the International Mycological Institute (IMI) under the accession number 375061.
- 5. Enzyme according to any of claims 1 to 4, characterized in that said enzyme comprises the amino acid sequence given in SEQ ID NO:1 or SEQ ID NO:3 or functional derivatives thereof.
- 6. Enzyme according to any of claims 1 to 5, characterized in that said enzyme is encoded by the DNA sequence given in SEQ ID NO:1 or SEQ ID NO:3 or functional derivatives or homologues thereof.
- 7. DNA molecule encoding an enzyme with phenolic acid esterase activity, characterized in that said DNA molecule comprises a DNA sequence as given in SEQ ID NO:1 or SEQ ID NO:3 or functional derivatives or homologues thereof.
- 8. DNA molecule encoding an enzyme according to any of claims 1 to 6, characterized in that said DNA molecule comprises a DNA sequence as given in SEQ ID NO:1 or SEQ ID NO:3 or functional derivatives or homologues thereof.
- 9. DNA molecule according to claim 7 or 8 further comprising vector sequence capable of expressing said enzyme in a procaryotic or eucaryotic host.
- 10. Transformed procaryotic cell or eucaryotic cell or organism comprising one or more DNA molecules according to claim 7 to 9.
- 11. Method for the production of an enzyme or enzyme preparation having phenolic acid esterase activity, characterized in that said enzyme is isolated from a cell or organism according to claim 10.
- 12. Enzyme preparation comprising enzyme according to any of claims 1 to 6 and/or obtainable by the method according to claim 11.
- 13. Enzyme preparation according to claim 12 comprising one or more further polysaccharide modifying and/or degrading enzymes.
- 14. Enzyme preparation according to claim 13, characterized in that said polysaccharide modifying and/or degrading enzyme is selected form the group comprising xylanase, arabinanase, a-L-arabinofuranosidase, endoglucanase, a-D glucuronidase, pectinase, acetyl esterase, mannanase, acetyl xylan esterase and other glycosyl hydrolases.
- 15. Enzyme preparation according to any of claims 11 to 14 comprising one or more further enzymes selected from the group comprising amylase, protease, a-galactosidase, phytase and lipase.
- 16. Use of the enzyme according to any of claims 1 to 6 and/or enzyme preparation according to any of claims 11 to 15 in a process for releasing or preparing phenolic acids from a substrate comprising phenolic acid moieties.
- 17. Use of the enzyme according to any of claims 1 to 6 and/or enzyme preparation according to any of claims 11 to 15 in the production of animal feed.
- 18. Use of the enzyme according to any of claims 1 to 6 and/or enzyme preparation according to any of claims 11 to 15 in the production of food.
- 9. Use of the enzyme enzyme according to any of claims 1 to 6 and/or enzyme preparation according to any of claims 11 to 15 in the production of paper.
- 20. Use of the enzyme according to any of claims 1 to 6 and/or enzyme preparation according to any of claims 11 to 15 in a process for bioconversion of plant material or ligno-cellulose wastes to sugars.
- 21. Use of the enzyme according to any of claims 1 to 6 and/or enzyme preparation according to any of claims 11 to 15 in crop plants to improve the digestibility of said plants for livestock.
- 22. Feed additive comprising the enzyme according to any of claims 1 to 6 and/or enzyme preparation according to any of claims 11 to 15.
- 23. Feed comprising the feed additive according to claim 22.
Priority Applications (8)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB9707540A GB2324302A (en) | 1997-04-14 | 1997-04-14 | Phenolic acid esterase |
| JP54346098A JP2001523090A (en) | 1997-04-14 | 1998-04-09 | Phenolic esterase and use thereof |
| BR9808554-9A BR9808554A (en) | 1997-04-14 | 1998-04-09 | Phenolic acid esterase and its use |
| AU76423/98A AU7642398A (en) | 1997-04-14 | 1998-04-09 | Phenolic acid esterase and use thereof |
| CA002286694A CA2286694A1 (en) | 1997-04-14 | 1998-04-09 | Phenolic acid esterase and use thereof |
| CN98804186A CN1255165A (en) | 1997-04-14 | 1998-04-09 | Phenolic acid esterase and use thereof |
| PCT/EP1998/002080 WO1998046768A2 (en) | 1997-04-14 | 1998-04-09 | Phenolic acid esterase and use thereof |
| EP98924104A EP0975768A2 (en) | 1997-04-14 | 1998-04-09 | Phenolic acid esterase and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB9707540A GB2324302A (en) | 1997-04-14 | 1997-04-14 | Phenolic acid esterase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| GB9707540D0 GB9707540D0 (en) | 1997-06-04 |
| GB2324302A true GB2324302A (en) | 1998-10-21 |
Family
ID=10810766
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| GB9707540A Withdrawn GB2324302A (en) | 1997-04-14 | 1997-04-14 | Phenolic acid esterase |
Country Status (8)
| Country | Link |
|---|---|
| EP (1) | EP0975768A2 (en) |
| JP (1) | JP2001523090A (en) |
| CN (1) | CN1255165A (en) |
| AU (1) | AU7642398A (en) |
| BR (1) | BR9808554A (en) |
| CA (1) | CA2286694A1 (en) |
| GB (1) | GB2324302A (en) |
| WO (1) | WO1998046768A2 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004009804A2 (en) * | 2002-07-18 | 2004-01-29 | Biocatalysts Limited | Feruloyl esterase and uses thereof |
| WO2006081825A1 (en) * | 2005-02-04 | 2006-08-10 | University Of Aarhus | A method for recycling important nutritional elements from waste |
| WO2009027638A1 (en) * | 2007-08-28 | 2009-03-05 | Biocatalysts Limited | Use of type c and d feruloyl esterases in the manufacture of biofuels |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1109912A1 (en) | 1998-09-04 | 2001-06-27 | University Of Georgia Research Foundation, Inc. | Phenolic acid esterases, coding sequences and methods |
| US6602700B1 (en) | 1998-09-04 | 2003-08-05 | University Of Georgia Research Foundation, Inc. | Phenolic acid esterases, coding sequences and methods |
| CN102365360A (en) * | 2009-03-24 | 2012-02-29 | 诺维信公司 | Polypeptide with acetylxylan esterase activity and polynucleotide encoding the polypeptide |
| CN102220298A (en) * | 2011-04-20 | 2011-10-19 | 中国科学院微生物研究所 | Ferulic acid esterase FaeI as well as coding gene and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2301103A (en) * | 1995-05-23 | 1996-11-27 | Danisco | Enzyme system comprising ferulic acid esterase activity |
-
1997
- 1997-04-14 GB GB9707540A patent/GB2324302A/en not_active Withdrawn
-
1998
- 1998-04-09 CN CN98804186A patent/CN1255165A/en active Pending
- 1998-04-09 EP EP98924104A patent/EP0975768A2/en not_active Withdrawn
- 1998-04-09 BR BR9808554-9A patent/BR9808554A/en not_active IP Right Cessation
- 1998-04-09 JP JP54346098A patent/JP2001523090A/en active Pending
- 1998-04-09 CA CA002286694A patent/CA2286694A1/en not_active Abandoned
- 1998-04-09 WO PCT/EP1998/002080 patent/WO1998046768A2/en not_active Application Discontinuation
- 1998-04-09 AU AU76423/98A patent/AU7642398A/en not_active Abandoned
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2301103A (en) * | 1995-05-23 | 1996-11-27 | Danisco | Enzyme system comprising ferulic acid esterase activity |
Non-Patent Citations (5)
| Title |
|---|
| Appl.Microbiol.Biotechnol. 1990,33,345-351 * |
| Applied and Environmental Micobiology 1992,58(11),3762-3766 * |
| Biochemical Society Transactions 1992,20,275S * |
| Enzyme Microb.Technol. 1992,14,875-88 * |
| Enzyme Microb.Technol. 1993,15,460-475 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004009804A2 (en) * | 2002-07-18 | 2004-01-29 | Biocatalysts Limited | Feruloyl esterase and uses thereof |
| WO2004009804A3 (en) * | 2002-07-18 | 2004-05-27 | Biocatalysts Ltd | Feruloyl esterase and uses thereof |
| WO2006081825A1 (en) * | 2005-02-04 | 2006-08-10 | University Of Aarhus | A method for recycling important nutritional elements from waste |
| WO2009027638A1 (en) * | 2007-08-28 | 2009-03-05 | Biocatalysts Limited | Use of type c and d feruloyl esterases in the manufacture of biofuels |
Also Published As
| Publication number | Publication date |
|---|---|
| WO1998046768A3 (en) | 1999-01-21 |
| BR9808554A (en) | 2000-05-23 |
| WO1998046768A2 (en) | 1998-10-22 |
| GB9707540D0 (en) | 1997-06-04 |
| AU7642398A (en) | 1998-11-11 |
| CA2286694A1 (en) | 1998-10-22 |
| JP2001523090A (en) | 2001-11-20 |
| EP0975768A2 (en) | 2000-02-02 |
| CN1255165A (en) | 2000-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8304212B2 (en) | Methods and compositions for degradation of lignocellulosic material | |
| CA2156048C (en) | Alkali-tolerant xylanases | |
| Garcia-Conesa et al. | The feruloyl esterase system of Talaromyces stipitatus: production of three discrete feruloyl esterases, including a novel enzyme, TsFaeC, with a broad substrate specificity | |
| Sanjivkumar et al. | Biosynthesis, purification and characterization of β-1, 4-xylanase from a novel mangrove associated actinobacterium Streptomyces olivaceus (MSU3) and its applications | |
| De La Mare et al. | Molecular and biochemical characterization of three GH62 α-L-arabinofuranosidases from the soil deuteromycete Penicillium funiculosum | |
| Crepin et al. | Production and characterization of the Talaromyces stipitatus feruloyl esterase FAEC in Pichia pastoris: identification of the nucleophilic serine | |
| WO2001042433A2 (en) | Talaromyces emersonii xylanase | |
| CN101909461A (en) | Polypeptides having acetylxylan esterase activity and polynucleotides encoding same | |
| WO1993024622A1 (en) | Mannanase enzymes, genes coding for them, methods for isolating the genes, and methods for bleaching lignocellulosic pulps | |
| RO116211B1 (en) | Arabinoxylan degrading polypeptide, recombinant dna encoding said polypeptide, preparation process and edible composition containing the same | |
| JP4267696B2 (en) | Enzyme having galactanase activity | |
| Aurilia et al. | Microbial carbohydrate esterases in cold adapted environments | |
| EP0896616A1 (en) | An enzyme with galactanase activity | |
| MERIVUORI et al. | Regulation of cellulase biosynthesis and secretion in fungi | |
| US6602700B1 (en) | Phenolic acid esterases, coding sequences and methods | |
| JPH09500785A (en) | Enzyme with acetylesterase activity | |
| Fadel et al. | Cellulases and animal feed production by solid-state fermentation by Aspergillus fumigatus NRCF-122 mutant | |
| KR100754479B1 (en) | New Fanibacilli sp. HＹ-8 strain and xylanase isolated from it | |
| GB2324302A (en) | Phenolic acid esterase | |
| US20140127754A1 (en) | Methods and compositions for degradation of lignocellulosic material | |
| Topakas et al. | Xylanases: characteristics, sources, production, and applications | |
| US6548283B1 (en) | Bacterial protein with xylanase activity | |
| WO2004009804A2 (en) | Feruloyl esterase and uses thereof | |
| CN101171335B (en) | AbfB-2 gene of penicillium funiculosum | |
| US11371032B2 (en) | Beta glucosidase with high glucose tolerance, high thermal stability and broad PH activity spectrum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| WAP | Application withdrawn, taken to be withdrawn or refused ** after publication under section 16(1) |