CN117448209A - Lactobacillus pentosus for efficiently degrading phosphorus phytate, fermentation preparation and application thereof - Google Patents
Lactobacillus pentosus for efficiently degrading phosphorus phytate, fermentation preparation and application thereof Download PDFInfo
- Publication number
- CN117448209A CN117448209A CN202311320596.9A CN202311320596A CN117448209A CN 117448209 A CN117448209 A CN 117448209A CN 202311320596 A CN202311320596 A CN 202311320596A CN 117448209 A CN117448209 A CN 117448209A
- Authority
- CN
- China
- Prior art keywords
- phosphorus
- phytate
- lactobacillus pentosus
- fermentation
- degrading
- 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.)
- Granted
Links
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 48
- 239000011574 phosphorus Substances 0.000 title claims abstract description 47
- 229910052698 phosphorus Inorganic materials 0.000 title claims abstract description 47
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 title claims abstract description 44
- 235000002949 phytic acid Nutrition 0.000 title claims abstract description 44
- 238000000855 fermentation Methods 0.000 title claims abstract description 42
- 230000004151 fermentation Effects 0.000 title claims abstract description 42
- 241000186684 Lactobacillus pentosus Species 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 230000000593 degrading effect Effects 0.000 title claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 15
- 239000007788 liquid Substances 0.000 claims description 27
- 239000001963 growth medium Substances 0.000 claims description 21
- 241000196324 Embryophyta Species 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 11
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 11
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 11
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 10
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 claims description 10
- 229910000360 iron(III) sulfate Inorganic materials 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 10
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 10
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 10
- 238000011218 seed culture Methods 0.000 claims description 9
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 239000001632 sodium acetate Substances 0.000 claims description 6
- 235000017281 sodium acetate Nutrition 0.000 claims description 6
- 235000019270 ammonium chloride Nutrition 0.000 claims description 5
- 235000013379 molasses Nutrition 0.000 claims description 5
- 239000001509 sodium citrate Substances 0.000 claims description 5
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 5
- 229920000742 Cotton Polymers 0.000 claims description 4
- 240000008042 Zea mays Species 0.000 claims description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- 235000012054 meals Nutrition 0.000 claims description 4
- 235000019779 Rapeseed Meal Nutrition 0.000 claims description 3
- 235000019764 Soybean Meal Nutrition 0.000 claims description 3
- 239000004456 rapeseed meal Substances 0.000 claims description 3
- 239000004455 soybean meal Substances 0.000 claims description 3
- 238000009472 formulation Methods 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- 235000015099 wheat brans Nutrition 0.000 claims 1
- 241001465754 Metazoa Species 0.000 abstract description 13
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 abstract description 12
- 239000000467 phytic acid Substances 0.000 abstract description 12
- 229940068041 phytic acid Drugs 0.000 abstract description 12
- 239000002994 raw material Substances 0.000 abstract description 11
- 239000003795 chemical substances by application Substances 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 3
- 239000006041 probiotic Substances 0.000 abstract description 3
- 230000000529 probiotic effect Effects 0.000 abstract description 3
- 235000018291 probiotics Nutrition 0.000 abstract description 3
- 230000008021 deposition Effects 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 239000000243 solution Substances 0.000 description 8
- 241000186660 Lactobacillus Species 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 7
- 229940039696 lactobacillus Drugs 0.000 description 7
- 230000015556 catabolic process Effects 0.000 description 6
- 238000006731 degradation reaction Methods 0.000 description 6
- 230000012010 growth Effects 0.000 description 6
- 238000012216 screening Methods 0.000 description 5
- 108010011619 6-Phytase Proteins 0.000 description 4
- 244000068988 Glycine max Species 0.000 description 4
- 235000010469 Glycine max Nutrition 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 4
- 230000002906 microbiologic effect Effects 0.000 description 4
- 229940085127 phytase Drugs 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000000877 morphologic effect Effects 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 241000186146 Brevibacterium Species 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 235000019728 animal nutrition Nutrition 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- -1 phytate calcium magnesium salts Chemical class 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000009604 anaerobic growth Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004134 energy conservation Methods 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 150000003017 phosphorus Chemical class 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 229940078499 tricalcium phosphate Drugs 0.000 description 1
- 229910000391 tricalcium phosphate Inorganic materials 0.000 description 1
- 235000019731 tricalcium phosphate Nutrition 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000005303 weighing Methods 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- Animal Husbandry (AREA)
- Physiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Mycology (AREA)
- Sustainable Development (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of microorganisms, and discloses lactobacillus pentosus for efficiently degrading phosphorus phytate, a fermentation preparation and application thereof. The invention provides a lactobacillus pentosus for efficiently degrading phosphorus phytate, which is deposited in the microorganism strain collection in Guangdong province at 2023, month 13 under the deposition number of GDMCC NO 63561. The lactobacillus pentosus for efficiently degrading the phytic acid can efficiently degrade the phytic acid; the fermentation efficiency is high, and the method can be used as a probiotic agent to treat raw materials or be applied to animal feeding, so that the utilization rate of phosphorus in plant raw materials is improved.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to lactobacillus pentosus for efficiently degrading phosphorus phytate, a fermentation preparation and application thereof.
Background
Most of the phosphorus in plant materials is organic phosphorus, accounting for about 85% of the total phosphorus, and exists mainly in the form of nucleic acid, phospholipids and phytate phosphorus. The content of the phytic acid phosphorus in different plant raw materials is greatly different, and especially the content of the phytic acid phosphorus in leguminous plants accounts for 1-5% of dry matters. Since phytate phosphorus affects the effectiveness of other mineral elements, chelate is formed which cannot be digested and absorbed by animals, resulting in reduced utilization of phosphorus by animals. In addition, phytate phosphorus affects the digestion and absorption of other nutrients such as proteins, starches and fats by animals. In order to solve the effect of phytate phosphorus on animal nutrition, the phytase method is generally adopted. Phytase is an enzyme capable of decomposing phytate phosphorus into phytate calcium magnesium salts and other soluble phytate salts. Besides the phytase method, inorganic phosphorus salt is added into the feed to supplement phosphorus source, ensure animal nutrition and improve animal growth performance. These methods are all relatively common and efficient at present, but the addition of phytase and inorganic phosphorus also increases the cost of the feed. In recent years, microbial degradation of phytic acid phosphorus is also a hot spot for research, and the microbial degradation of phytic acid phosphorus is a method for effectively improving the utilization rate of phosphorus in feed mainly through acid production or enzyme production.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide lactobacillus pentosus for efficiently degrading phosphorus phytate, a fermentation preparation and application thereof.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect, the invention provides a lactobacillus pentosus strain with a deposit number of GDMCC NO. 63561, which can degrade phosphorus phytate efficiently. The strain is named as lactobacillus pentosus Lactiplantibacillus pentosus BOEN-1418 and is deposited with the Guangdong province microbiological bacterial collection center (GDMCC) on the 13 th month of 2023, and the deposit address is: guangzhou City first middle road No. 100 college No. 59 building No. 5 Guangdong province microbiological institute.
The lactobacillus pentosus for efficiently degrading the phytate is derived from a fermented soybean sample, belongs to a natural and safe strain, does not have any side effect, has high safety when applied to animal feeds, belongs to lactobacillus, can be well utilized by animals, can be planted in the intestines of the animals, and can inhibit the growth of harmful bacteria. The culture and fermentation conditions are simple, the genetic property is stable, and the liquid fermentation preparation can efficiently degrade the phytate phosphorus; the fermentation efficiency is high, the lactobacillus is efficiently enriched in a short time, the lactobacillus can be used as a probiotic agent for treating raw materials or applied to animal feeding, the phytic acid phosphorus is efficiently degraded, and the utilization rate of the plant raw material phosphorus is improved.
In a second aspect, the invention provides a fermentation preparation prepared from the lactobacillus pentosus for efficiently degrading phosphorus phytate.
In a third aspect, the invention provides a method for preparing the fermentation preparation, comprising the following steps:
(1) Inoculating the lactobacillus pentosus for efficiently degrading the phytate phosphorus into a liquid seed culture medium for culture to obtain seed liquid;
(2) Inoculating the seed liquid into a fermentation culture medium according to the inoculum size of 1-4% by volume, stirring, and fermenting and culturing to obtain the seed liquid.
The lactobacillus pentosus for efficiently degrading the phytate phosphorus has the advantages of simple fermentation operation, energy conservation and cost reduction, and is easier for industrial production.
As a preferred embodiment of the preparation method of the present invention, in the step (1), the seed medium has a formula of: 10g/L to 40g/L of glucose, 5g/L to 12g/L of yeast powder, 0.5g/L to 5g/L of magnesium sulfate heptahydrate, 0.5g/L to 1.5g/L of ferric sulfate, 0.5g/L to 3g/L of potassium dihydrogen phosphate and pH value of 6.0 to 7.0.
As a preferred embodiment of the production method of the present invention, in the step (1), the temperature of the culture is 35 to 42℃and the time is 18 to 24 hours.
As a preferred embodiment of the preparation method of the present invention, in the step (2), the formulation of the fermentation medium is: 30g/L to 100g/L of molasses, 8g/L to 15g/L of yeast powder, 5g/L to 10.5g/L of ammonium chloride, 0.5g/L to 1.5g/L of ferric sulfate, 0.7g/L to 1.4g/L of magnesium sulfate heptahydrate, 2g/L to 5g/L of sodium acetate, 0.5g/L to 2g/L of sodium citrate, 1g/L to 2.5g/L of monopotassium phosphate and pH value of 6.0 to 7.0.
As a preferred embodiment of the preparation method of the present invention, in the step (2), the stirring rotation speed is 50r/min to 100r/min; the temperature of the fermentation culture is 35-42 ℃ and the time is 24-48 h.
In a fourth aspect, the invention provides the use of lactobacillus pentosus for degrading phosphorus phytate in plant material.
In a fifth aspect, the invention provides the use of the fermentation broth for degrading phosphorus phytate in a plant material.
As a preferred embodiment of the application of the present invention, the plant material is at least one of soybean meal, rapeseed meal, cotton meal, corn and bran.
Compared with the prior art, the invention has the beneficial effects that:
the lactobacillus pentosus for efficiently degrading the phytate is derived from a fermented soybean sample, belongs to a natural and safe strain, does not have any side effect, has high safety when applied to animal feeds, belongs to lactobacillus, can be well utilized by animals, can be planted in the intestines of the animals, and can inhibit the growth of harmful bacteria. The culture and fermentation conditions are simple, the genetic property is stable, and the liquid fermentation preparation can efficiently degrade the phytate phosphorus; the fermentation efficiency is high, the lactobacillus is efficiently enriched in a short time, the lactobacillus can be used as a probiotic agent for treating raw materials or applied to animal feeding, the phytic acid phosphorus is efficiently degraded, and the utilization rate of the plant raw material phosphorus is improved.
Drawings
FIG. 1 is a colony morphology of strain BOEN-1418;
FIG. 2 shows the strain BOEN-1418 under a microscope.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to the following specific examples. It will be appreciated by persons skilled in the art that the specific embodiments described herein are for purposes of illustration only and are not intended to be limiting.
The test methods used in the examples are conventional methods unless otherwise specified; the materials, reagents and the like used, unless otherwise specified, are all commercially available.
Example 1: strain screening and identification
(1) Strain screening
Strain BOEN-1418 strain was isolated from Huang Mingyuan samples taken at day 5 of 9 of 2022.
The BOEN-1418 strain is obtained by separating and purifying traditional fermented soybean samples from farmhouse in Qingyuan city of Guangdong province by adopting a gradient dilution coating plate method and a streak culture method, and the pure cultured strain is preserved in a refrigerator at 4 ℃. The method comprises the following specific steps:
phosphate-solubilizing separation screening medium: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast powder, 20g/L of glucose, 2g/L of diammonium hydrogen citrate, 0.58g/L of magnesium sulfate heptahydrate, 3.12g/L of sodium acetate, 1g/L of tween 80, 0.25g/L of manganese sulfate monohydrate, 5g/L of tricalcium phosphate, 20g/L of agar powder, supplementing distilled water to 1000ml, regulating the pH value of a culture medium to 6.7-6.9, and sterilizing at 115 ℃ for 30min.
And (3) separating and purifying: weighing 10.0g fermented soybean sample, dissolving in 90ml sterile water, shaking at 25deg.C and 180rpm for 30min, and sequentially diluting to 10 -7 . Respectively draw 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 100ul of gradient diluent is connected into phosphate solubilizingUniformly coating on a screening culture medium plate, reversely culturing for 48 hours in a 37 ℃ incubator, picking out bacterial colonies in different forms capable of generating clear transparent halation, continuously culturing for 24 hours in the 37 ℃ incubator on a phosphorus-dissolving separation screening culture medium plate, microscopic examination of bacterial colony morphology of each bacterial colony, picking out rod-shaped bacterial colonies, and repeatedly marking, separating and purifying until single bacterial colonies grow on the phosphorus-dissolving separation culture medium plate.
(2) Identification of strains
The screened strain is tested for morphological characteristics, physiological and biochemical characteristics and other indexes, and the method is concretely as follows:
morphological characteristics: BOEN-1418 strain was streaked on solid seed medium and cultured overnight at 30℃to observe colony morphology and color. The strain BOEN-1418 has good growth state on a solid seed culture medium, colony morphology is as shown in figure 1, and the colony is round, opaque, milky white, convex, neat in edge, moist in surface and easy to pick up. The morphology of the cells is observed under a microscope as shown in FIG. 2: the microscopic examination is carried out on Brevibacterium, single or short-chain arrangement, the Brevibacterium has blunt ends, thick capsules are arranged around the thalli, and gram positive is carried out.
Physiological and biochemical characteristics: the strain BOEN-1418 has the advantages of facultative anaerobic growth, starch hydrolysis, positive contact enzyme reaction, negative oxidase and lecithin enzyme reaction, good growth on nitrogen-free culture medium, and good growth in the environment of 10-45 ℃.
(3) Molecular biological identification
Genomic DNA of strain BOEN-1418 was extracted according to the instructions of TaKaRa MiniBEST Bacteria Genomic DNA Extraction Kit Ver.3.0 kit and the 16S rRNA gene sequences were amplified using bacterial universal primers 27-F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492-R (5'-GGTTACCTTGTTACGACTT-3') using the extracted genomic DNA as template. The PCR products were sequenced by Shangya bioengineering (Shanghai) Inc., and the sequencing results were compared to the GenBank database in NCBI.
The genome DNA of the strain BOEN-1418 is used as a template to amplify to obtain a 16S rRNA gene segment with the length of 1435bp (SEQ ID NO. 1), and the sequencing result is compared with sequences in GenBank, so that the result shows that the BOEN-1418 belongs to Lactobacillus and has 99 percent of similarity with the 16S rRNA sequences of different strains of the typical strain Lactiplantibacillus pentosus of the genus in a database, and the strain is identified as Lactobacillus pentosus (Lactiplantibacillus pentosus) by combining with morphological characteristics and the like of the strain.
Thus, the strain was named Lactobacillus pentosus Lactiplantibacillus pentosus BOEN-1418, deposited at the Guangdong province microbiological bacterial collection center (GDMCC) at month 13 of 2023, accession number: guangzhou city, first, middle road 100, building 59, building 5, guangdong province microbiological institute, deposit number: GDMCCNO 63561.
Example 2: liquid fermentation of lactobacillus pentosus BOEN-1418
(1) Culture medium
Seed culture medium: 35g/L glucose, 10g/L yeast powder, 3.6g/L magnesium sulfate heptahydrate, 0.8g/L ferric sulfate, 2.5g/L potassium dihydrogen phosphate and pH 6.0-7.0, and is prepared by distilled water.
Liquid fermentation medium: 88g/L molasses, 15g/L yeast powder, 8g/L ammonium chloride, 1g/L ferric sulfate, 0.7g/L magnesium sulfate heptahydrate, 3.6g/L sodium acetate, 0.5g/L sodium citrate, 1.8g/L potassium dihydrogen phosphate and pH of 6.0-7.0 are prepared by distilled water.
The lactobacillus pentosus BOEN-1418 liquid fermentation comprises the following steps:
selecting preserved lactobacillus pentosus BOEN-1418, inoculating into liquid seed culture medium, and culturing at 37deg.C for 18 to obtain seed solution; inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 4%, and fermenting for 30 hours at 37 ℃ and stirring speed of 100r/min to obtain a fermentation liquid preparation.
Example 3: liquid fermentation of lactobacillus pentosus BOEN-1418
(1) Culture medium
Seed culture medium: 10g/L of glucose, 5g/L of yeast powder, 0.5g/L of magnesium sulfate heptahydrate, 0.5g/L of ferric sulfate, 0.5g/L of potassium dihydrogen phosphate and pH of 6.0-7.0, and is prepared by distilled water.
Liquid fermentation medium: 30g/L of molasses, 8g/L of yeast powder, 10.5g/L of ammonium chloride, 0.5g/L of ferric sulfate, 1g/L of magnesium sulfate heptahydrate, 2g/L of sodium acetate, 1.2g/L of sodium citrate, 1g/L of potassium dihydrogen phosphate and pH of 6.0-7.0, and is prepared by distilled water.
The lactobacillus pentosus BOEN-1418 liquid fermentation comprises the following steps:
selecting preserved lactobacillus pentosus BOEN-1418, inoculating into a liquid seed culture medium, and culturing at 42 ℃ for 24 hours to obtain seed liquid; inoculating the seed solution into a fermentation culture medium according to the inoculation amount of 3%, and fermenting for 48 hours at the temperature of 42 ℃ and the stirring speed of 80r/min to obtain a fermentation liquid preparation.
Example 4: liquid fermentation of lactobacillus pentosus BOEN-1418
(1) Culture medium
Seed culture medium: 40g/L glucose, 12g/L yeast powder, 5g/L magnesium sulfate heptahydrate, 1.5g/L ferric sulfate, 3g/L potassium dihydrogen phosphate, pH 6.0-7.0, and distilled water.
Liquid fermentation medium: 100g/L of molasses, 12g/L of yeast powder, 5g/L of ammonium chloride, 1.5g/L of ferric sulfate, 1.4g/L of magnesium sulfate heptahydrate, 5g/L of sodium acetate, 2g/L of sodium citrate and 2.5g/L of potassium dihydrogen phosphate, and the pH value is 6.0-7.0, and the compound fertilizer is prepared by distilled water.
The lactobacillus pentosus BOEN-1418 liquid fermentation comprises the following steps:
selecting preserved lactobacillus pentosus BOEN-1418, inoculating into a liquid seed culture medium, and culturing at 35 ℃ for 18h to obtain seed liquid; inoculating 1% seed solution into fermentation medium, and fermenting at 35deg.C with stirring speed of 50r/min for 24 hr to obtain fermentation broth preparation.
Test example:
the fermentation broth preparations prepared in examples 2 to 4 were taken, and the effect of efficiently degrading phosphorus phytate was examined by applying the liquid preparation to a plant material and detecting the change of phosphorus phytate during fermentation.
The fermentation broth preparations prepared in examples 2 to 4 are used for plant raw materials such as soybean meal, rapeseed meal, cotton meal, corn, bran and the like, and the addition amount is based on the bacterial content of the raw materials: 10 6 cfu/g, water content 38%, temperature 37 ℃, fermenting for 24 hours, measuring the content of the phytic acid, and calculating the degradation rate of the phytic acid, wherein the comparison is made by taking the preparation without adding fermentation liquor. The phytic acid phosphorus content is determined by adopting a precipitation digestion method, and the degradation rate of the phytic acid phosphorus is=the raw materialThe amount of the degraded phosphorus phytate/total phosphorus phytate of the raw materials is 100%.
TABLE 1 degradation rate of plant material phosphorus phytate (%)
| Group of | Bean pulp | Vegetable meal | Cotton seed cake | Corn | Bran |
| Example 2 | 96.36±1.21 | 89.87±0.84 | 84.98±1.09 | 97.34±1.22 | 83.38±0.91 |
| Example 3 | 95.67±1.89 | 89.43±1.55 | 85.36±2.13 | 98.02±1.75 | 82.25±0.22 |
| Example 4 | 96.43±1.25 | 87.94±2.01 | 86.54±2.32 | 98.70±2.60 | 81.48±1.35 |
As shown in Table 1, the degradation rate of the raw material phytate phosphorus of the lactobacillus pentosus Lactiplantibacillus pentosus BOEN-1418 fermentation broth preparation prepared in the added examples 2-4 is high.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted equally without departing from the spirit and scope of the technical solution of the present invention.
Claims (10)
1. A lactobacillus pentosus for efficiently degrading phytate phosphorus is characterized in that the storage number is GDMCCNO 63561.
2. A fermented preparation prepared from the phosphor phytate-degrading lactobacillus pentosus of claim 1.
3. A method of preparing a fermented preparation according to claim 2, comprising the steps of:
(1) Inoculating the lactobacillus pentosus capable of efficiently degrading the phosphorus phytate in the liquid seed culture medium for culture to obtain seed liquid;
(2) Inoculating the seed liquid into a fermentation culture medium according to the inoculum size of 1-4% by volume, stirring, and fermenting and culturing to obtain the seed liquid.
4. The method of claim 3, wherein in step (1), the seed medium is formulated as follows: 10g/L to 40g/L of glucose, 5g/L to 12g/L of yeast powder, 0.5g/L to 5g/L of magnesium sulfate heptahydrate, 0.5g/L to 1.5g/L of ferric sulfate, 0.5g/L to 3g/L of potassium dihydrogen phosphate and pH value of 6.0 to 7.0.
5. The method according to claim 3, wherein in the step (1), the temperature of the culture is 35 to 42℃and the time is 18 to 24 hours.
6. A method of preparation according to claim 3, wherein in step (2) the fermentation medium is formulated as: 30g/L to 100g/L of molasses, 8g/L to 15g/L of yeast powder, 5g/L to 10.5g/L of ammonium chloride, 0.5g/L to 1.5g/L of ferric sulfate, 0.7g/L to 1.4g/L of magnesium sulfate heptahydrate, 2g/L to 5g/L of sodium acetate, 0.5g/L to 2g/L of sodium citrate, 1g/L to 2.5g/L of monopotassium phosphate and pH value of 6.0 to 7.0.
7. A method according to claim 3, wherein in step (2), the stirring speed is 50r/min to 100r/min; the temperature of the fermentation culture is 35-42 ℃ and the time is 24-48 h.
8. Use of lactobacillus pentosus as claimed in claim 1 for degrading phytate phosphorus in plant material.
9. Use of the fermented formulation of claim 2 for degrading phytate phosphorus in plant material.
10. The use according to claim 8 or 9, wherein the plant material is at least one of soybean meal, rapeseed meal, cotton meal, corn, wheat bran.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311320596.9A CN117448209B (en) | 2023-10-12 | 2023-10-12 | Lactobacillus pentosus for efficiently degrading phosphorus phytate, fermentation preparation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311320596.9A CN117448209B (en) | 2023-10-12 | 2023-10-12 | Lactobacillus pentosus for efficiently degrading phosphorus phytate, fermentation preparation and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117448209A true CN117448209A (en) | 2024-01-26 |
| CN117448209B CN117448209B (en) | 2024-08-13 |
Family
ID=89584585
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311320596.9A Active CN117448209B (en) | 2023-10-12 | 2023-10-12 | Lactobacillus pentosus for efficiently degrading phosphorus phytate, fermentation preparation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117448209B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118562624A (en) * | 2024-06-07 | 2024-08-30 | 播恩集团股份有限公司 | Monascus purpureus and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104012797A (en) * | 2014-05-31 | 2014-09-03 | 湖北省农业科学院畜牧兽医研究所 | Early-stage compound feed for broiler chickens and preparation method thereof |
| CN116083273A (en) * | 2022-09-05 | 2023-05-09 | 新希望六和股份有限公司 | Lactobacillus plantarum NHE-LpE15 and application thereof |
| US20230242890A1 (en) * | 2020-05-22 | 2023-08-03 | Qingdao Vland Biotech Group Co., Ltd. | Phytase mutant |
-
2023
- 2023-10-12 CN CN202311320596.9A patent/CN117448209B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104012797A (en) * | 2014-05-31 | 2014-09-03 | 湖北省农业科学院畜牧兽医研究所 | Early-stage compound feed for broiler chickens and preparation method thereof |
| US20230242890A1 (en) * | 2020-05-22 | 2023-08-03 | Qingdao Vland Biotech Group Co., Ltd. | Phytase mutant |
| CN116083273A (en) * | 2022-09-05 | 2023-05-09 | 新希望六和股份有限公司 | Lactobacillus plantarum NHE-LpE15 and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| ILARIA DE PASQUALE等: ""Nutritional and functional effects of the lactic acid bacteria fermentation on gelatinized legume flours"", 《INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY》, vol. 2020, no. 316, 2 March 2020 (2020-03-02), pages 1 - 12 * |
| 王赫等: ""不同发酵时间对乳酸菌发酵饲料中主要营养物质主要营养物质、乳酸菌和乳酸含量的影响"", 《中国家禽》, vol. 39, no. 10, 25 May 2017 (2017-05-25), pages 27 - 31 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118562624A (en) * | 2024-06-07 | 2024-08-30 | 播恩集团股份有限公司 | Monascus purpureus and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117448209B (en) | 2024-08-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102936572B (en) | Acid resistance and high temperature resistance saccharomyces cerevisiae and applications thereof | |
| CN115287202B (en) | A Kluyveromyces marx strain and its application in the preparation of functional feed | |
| CN101074426B (en) | Method for producing bean-dregs feed containing conjugated linolic acid by plant lactobacillin fermentation | |
| CN109517761A (en) | The bacillus licheniformis of cellulase-producing, its microbial fermentation preparation and its application | |
| CN117448209B (en) | Lactobacillus pentosus for efficiently degrading phosphorus phytate, fermentation preparation and application thereof | |
| Li et al. | A β-glucosidase-producing M-2 strain: Isolation from cow dung and fermentation parameter optimization for flaxseed cake | |
| CN115181707A (en) | Bacillus subtilis and liquid-solid two-phase fermentation method thereof | |
| CN109423466B (en) | Compound fermentation inoculant and application thereof | |
| CN117603889B (en) | Bacillus subtilis for producing acid protease for feed and application thereof | |
| CN114468119A (en) | A kind of highland barley processing by-product compound biological feed and preparation method thereof | |
| CN117187079B (en) | Monilinia purpurea MN114 and application thereof | |
| CN104087529A (en) | Bacillus pumilus JY2 and applications thereof | |
| CN111187744A (en) | High-density industrial fermentation medium for stratospheric bacillus and fermentation method thereof | |
| CN114015607B (en) | Bacillus amyloliquefaciens for high yield of 5-methyltetrahydrofolic acid and application thereof | |
| CN113355265B (en) | Lactobacillus plantarum and application thereof | |
| CN111040969B (en) | Compound lactobacillus agent and application thereof in buffalo silage | |
| CN115595275A (en) | Monascus strain capable of highly yielding monascus pigment and application thereof | |
| CN113875975A (en) | Fermentation process for preparing metagens by using wheat processing byproducts | |
| CN113528599A (en) | Production method of efficient chelating enzyme peptide | |
| CN118308433B (en) | Application of bacillus subtilis BS-22 strain in fermentation of rapeseed meal | |
| CN114350534B (en) | Saccharomyces cerevisiae, fattening sheep biological feed and preparation method and application thereof | |
| AU2014229409A1 (en) | Extraction of nitrogen from organic materials through ammonification by mixed bacterial populations | |
| CN117904009B (en) | Bacillus subtilis applicable to non-grain bio-based carbon source and fermentation production method thereof | |
| CN115261356B (en) | Phytase yield increasing agent produced by clay fermentation and preparation method thereof | |
| CN112899197B (en) | Compound microbial agent, preparation thereof and application thereof in feed additive |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |