CN119614547A - Biological compound enzyme preparation and preparation method thereof - Google Patents
Biological compound enzyme preparation and preparation method thereof Download PDFInfo
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- CN119614547A CN119614547A CN202411749330.0A CN202411749330A CN119614547A CN 119614547 A CN119614547 A CN 119614547A CN 202411749330 A CN202411749330 A CN 202411749330A CN 119614547 A CN119614547 A CN 119614547A
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 210
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 210
- 238000002360 preparation method Methods 0.000 title claims abstract description 32
- 150000001875 compounds Chemical class 0.000 title description 12
- 239000012752 auxiliary agent Substances 0.000 claims abstract description 36
- 239000008187 granular material Substances 0.000 claims abstract description 33
- 238000000034 method Methods 0.000 claims abstract description 31
- 239000002131 composite material Substances 0.000 claims abstract description 28
- 239000002245 particle Substances 0.000 claims abstract description 20
- 239000003381 stabilizer Substances 0.000 claims abstract description 18
- 230000008569 process Effects 0.000 claims abstract description 14
- 229940088598 enzyme Drugs 0.000 claims description 197
- 239000008188 pellet Substances 0.000 claims description 48
- 239000010410 layer Substances 0.000 claims description 26
- 239000011241 protective layer Substances 0.000 claims description 16
- 239000004367 Lipase Substances 0.000 claims description 13
- 102000004882 Lipase Human genes 0.000 claims description 13
- 108090001060 Lipase Proteins 0.000 claims description 13
- 235000019421 lipase Nutrition 0.000 claims description 13
- 108091005658 Basic proteases Proteins 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 10
- 238000001125 extrusion Methods 0.000 claims description 10
- 239000004382 Amylase Substances 0.000 claims description 9
- 108010065511 Amylases Proteins 0.000 claims description 9
- 102000013142 Amylases Human genes 0.000 claims description 9
- 108010029541 Laccase Proteins 0.000 claims description 9
- 235000019418 amylase Nutrition 0.000 claims description 9
- 239000001110 calcium chloride Substances 0.000 claims description 9
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 9
- 229920000168 Microcrystalline cellulose Polymers 0.000 claims description 8
- 239000008108 microcrystalline cellulose Substances 0.000 claims description 8
- 229940016286 microcrystalline cellulose Drugs 0.000 claims description 8
- 235000019813 microcrystalline cellulose Nutrition 0.000 claims description 8
- 229940051841 polyoxyethylene ether Drugs 0.000 claims description 8
- 229920000056 polyoxyethylene ether Polymers 0.000 claims description 8
- 238000005507 spraying Methods 0.000 claims description 8
- 108010059892 Cellulase Proteins 0.000 claims description 7
- 229940106157 cellulase Drugs 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 7
- 230000007935 neutral effect Effects 0.000 claims description 7
- 102100032487 Beta-mannosidase Human genes 0.000 claims description 6
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 6
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims description 6
- 108010059820 Polygalacturonase Proteins 0.000 claims description 6
- 238000000889 atomisation Methods 0.000 claims description 6
- 108010055059 beta-Mannosidase Proteins 0.000 claims description 6
- 238000005422 blasting Methods 0.000 claims description 6
- 239000007771 core particle Substances 0.000 claims description 6
- 108010093305 exopolygalacturonase Proteins 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 5
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 claims description 5
- 239000004375 Dextrin Substances 0.000 claims description 5
- 229920001353 Dextrin Polymers 0.000 claims description 5
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 5
- 239000000654 additive Substances 0.000 claims description 5
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 5
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 5
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 5
- 235000019425 dextrin Nutrition 0.000 claims description 5
- 150000002191 fatty alcohols Chemical class 0.000 claims description 5
- 239000000600 sorbitol Substances 0.000 claims description 5
- 239000002202 Polyethylene glycol Substances 0.000 claims description 4
- 230000002572 peristaltic effect Effects 0.000 claims description 4
- 229920001223 polyethylene glycol Polymers 0.000 claims description 4
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 3
- 239000003921 oil Substances 0.000 claims description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 claims description 2
- 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 2
- 229930195725 Mannitol Natural products 0.000 claims description 2
- 230000000996 additive effect Effects 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 239000000594 mannitol Substances 0.000 claims description 2
- 235000010355 mannitol Nutrition 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- -1 sodium alkyl sulfonate Chemical class 0.000 claims description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 2
- 229960001763 zinc sulfate Drugs 0.000 claims description 2
- 239000007884 disintegrant Substances 0.000 claims 2
- 238000005469 granulation Methods 0.000 claims 2
- 230000003179 granulation Effects 0.000 claims 2
- 229940113116 polyethylene glycol 1000 Drugs 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 29
- 238000002156 mixing Methods 0.000 abstract description 18
- 239000006187 pill Substances 0.000 abstract description 15
- 238000004519 manufacturing process Methods 0.000 abstract description 9
- 239000000428 dust Substances 0.000 abstract description 7
- 238000009776 industrial production Methods 0.000 abstract description 5
- 230000014759 maintenance of location Effects 0.000 abstract description 5
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- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 230000001737 promoting effect Effects 0.000 abstract 1
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- 239000000843 powder Substances 0.000 description 9
- 238000001514 detection method Methods 0.000 description 7
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- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 5
- 229920001661 Chitosan Polymers 0.000 description 5
- GUJOJGAPFQRJSV-UHFFFAOYSA-N dialuminum;dioxosilane;oxygen(2-);hydrate Chemical compound O.[O-2].[O-2].[O-2].[Al+3].[Al+3].O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O GUJOJGAPFQRJSV-UHFFFAOYSA-N 0.000 description 5
- 230000003100 immobilizing effect Effects 0.000 description 5
- 229910052901 montmorillonite Inorganic materials 0.000 description 5
- 239000000661 sodium alginate Substances 0.000 description 5
- 235000010413 sodium alginate Nutrition 0.000 description 5
- 229940005550 sodium alginate Drugs 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 229920000858 Cyclodextrin Polymers 0.000 description 4
- 239000003513 alkali Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 3
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000001116 FEMA 4028 Substances 0.000 description 2
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 2
- 108010093096 Immobilized Enzymes Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 description 2
- 235000011175 beta-cyclodextrine Nutrition 0.000 description 2
- 229960004853 betadex Drugs 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
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- 238000004132 cross linking Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
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- 239000007788 liquid Substances 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 1
- 235000012766 Cannabis sativa ssp. sativa var. sativa Nutrition 0.000 description 1
- 235000012765 Cannabis sativa ssp. sativa var. spontanea Nutrition 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 235000009120 camo Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
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- 210000003491 skin Anatomy 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
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- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
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- 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
Landscapes
- Enzymes And Modification Thereof (AREA)
Abstract
The invention belongs to the technical field of enzyme engineering, and particularly relates to a composite enzyme particle and a preparation method thereof. The enzyme granule comprises an enzyme pill core and an auxiliary agent layer, wherein the enzyme pill core is used as a core part, the auxiliary agent layer is coated on the outer layer of the enzyme pill core, and the enzyme pill core comprises an enzyme component and a stabilizer. The improved composite enzyme particles improve the storage stability of the enzyme, store under the same condition, improve the retention rate of the enzyme activity by 20-30%, improve the mixing uniformity of the enzyme by 20% after shaking with the same intensity, improve the stress resistance in the use process, store under the same condition after dissolving with high pH, prolong the half life of the enzyme by 30-50%, and in addition, can reduce the dust and labor intensity in the use process and the influence of human factors on the production, and is beneficial to promoting the automation of industrial production, thereby improving the stability of the production process and improving the product quality.
Description
Technical field:
the invention belongs to the technical field of enzyme engineering, and particularly relates to a high-efficiency biological compound enzyme preparation granule and a preparation method thereof.
Technical background:
At present, biological enzymes are widely used in textile degumming and refining treatment, and can greatly improve the production efficiency. Compared with chemical treatment, the use of biological enzyme has unique treatment effect particularly on the degumming process of cotton, hemp and the like and the textile finishing process of jeans and the like. At present, most of industrially used enzymes are added in the form of liquid single enzyme or compound enzyme powder, and the liquid compound enzyme has poor stability and is unfavorable for storage and transportation. The components such as enzymes and fillers in the solid composite enzyme powder are not uniform in specific gravity and fineness, so that materials with different specific gravities and fineness are easy to separate and even layer in the process of carrying or transporting, and the mixing uniformity of the composite enzyme preparation is affected, so that the using effect is affected. The powder complex enzyme is easy to generate dust during the weighing and using and adding processes, and the contact or inhalation of people can cause contact allergic reaction, for example, if the contact or inhalation is excessive for a long time, the contact or inhalation can cause harm to human skin, eyes, nasal mucosa and tissues thereof, and the health is affected.
In the aspect of stress resistance and use effect of the enzyme, the stress resistance of the enzyme is improved by a genetic engineering means at present so as to improve the application effect of the enzyme, but the improvement is also achieved, but the genetic engineering aspect has high cost for enzyme modification, long research and development period, unsatisfactory effect after modification and the like, and the requirements of low cost and urgent need of the current industry cannot be met.
The technical staff in the field aim to develop a stable and efficient enzyme granule and a preparation process thereof so as to meet the requirements of high efficiency, simplicity and safety of industrial production, realize low dust or no dust in workshops, reduce the labor intensity of people and the influence of human factors on the production, promote the automation of the industrial production, and improve the stability of the production process so as to improve the product quality.
The invention comprises the following steps:
the invention aims to provide stable, efficient and environment-friendly composite enzyme particles and a preparation method thereof,
The invention provides a stable and efficient composite enzyme particle, which comprises an enzyme pellet core and an auxiliary agent layer, wherein the enzyme pellet core is used as a core part, the auxiliary agent layer is coated on the outer layer of the enzyme pellet core, and the enzyme pellet core comprises an enzyme component and a stabilizer;
Further, the weight ratio of the enzyme pellet core to the auxiliary agent layer is 4:1;
further, the weight part ratio of the enzyme component and the stabilizer in the enzyme pellet core is 40-85:10-40, and preferably, the weight part ratio of the enzyme component and the stabilizer is 60-65:35-40.
Further, the enzyme component is selected from 2 or more of the following enzyme classes alkaline protease, alkaline pectinase, alkaline lipase, neutral cellulase, xylanase, laccase, amylase and mannanase;
preferably, the enzyme component comprises, by weight, 1-5 parts of alkaline protease, 10-20 parts of alkaline pectase, 6-10 parts of alkaline lipase, 30-40 parts of neutral cellulase, 20-30 parts of xylanase, 5-10 parts of laccase, 5-10 parts of amylase and 2-5 parts of mannanase;
Further, the stabilizer is selected from 2 or more of sorbitol, glucose, dextrin, trehalose, mannitol, calcium chloride, copper sulfate and zinc sulfate;
Preferably, the stabilizer comprises the following components, by weight, 30-40 parts of sorbitol, 35-40 parts of dextrin, 20-25 parts of trehalose, 4.5-6 parts of calcium chloride and 0.3-0.5 part of copper sulfate;
further, the auxiliary agent layer comprises one or more of polyethylene glycol, sodium alkyl sulfonate, alkylphenol ethoxylates, fatty alcohol ethoxylates and tween;
Preferably, the auxiliary agent layer comprises 60 parts by weight of C12-C14 fatty alcohol polyoxyethylene ether, 1000 30 parts by weight of polyethylene glycol and 10 parts by weight of alkylphenol polyoxyethylene ether.
The second technical scheme provided by the invention is a preparation method of the composite enzyme particles, which comprises the steps of enzyme immobilization treatment, enzyme pellet core preparation and addition of an auxiliary agent layer, and specifically comprises the following steps:
(1) The immobilization treatment of the enzyme is as follows:
Immobilizing all or part of the enzyme components in the enzyme pellet core, immobilizing the enzyme to be immobilized by adopting an immobilization method commonly used in the field;
further, the immobilization is to add enzyme into an immobilization reagent, stir and mix, and carry out embedding, adsorption or crosslinking immobilization treatment at a certain pH and temperature;
Further, the enzyme to be immobilized is alkaline protease, alkaline pectase, alkaline lipase, amylase and laccase;
The immobilized reagent is one or more selected from cyclodextrin, carboxymethyl starch, sodium carboxymethyl cellulose, sodium alginate, calcium chloride, silicon dioxide, polyvinyl alcohol, chitosan, montmorillonite and calcium phosphate;
still further, the immobilizing agent includes, but is not limited to, at least one of sodium alginate, calcium chloride, cyclodextrin, chitosan, CMC, montmorillonite, etc.;
Preferably, the alkaline protease is immobilized by embedding sodium alginate and calcium chloride so as to reduce the hydrolysis of the alkaline protease;
preferably, the alkaline lipase is subjected to embedding treatment by adopting beta-cyclodextrin, so that the catalytic activity and stress resistance of the alkaline lipase are improved;
preferably, the alkaline pectinase is crosslinked and fixed by chitosan so as to improve alkali resistance and heat resistance;
Preferably, the amylase or laccase is immobilized by montmorillonite adsorption to improve alkali resistance of the enzyme.
(2) The preparation method of the enzyme pill core comprises the following steps:
mixing all enzyme components and stabilizing agents in the enzyme pill core according to the weight ratio of 40-85:10-40 to obtain a mixture, adding microcrystalline cellulose disintegrating agent according to the weight ratio of 90-95:5-10 of the mixture, uniformly mixing, preparing a wet enzyme pill core by adopting a wet rotary extrusion granulating method, and then drying the wet enzyme pill core in a fluidized bed;
further, the weight ratio of the materials in the granulating process of the enzyme pill core is 1:5-2:5;
further, the temperature of the spun extruded material is <70 ℃, preferably <60 ℃, more preferably <50 ℃;
further, extruding and granulating by using a 30-40 mesh net, and controlling the diameter of the extruded strip to be 0.3-0.5mm;
Further, adding the extruded strip into a shot blasting machine, and blasting at a rotating speed of 40-120rpm for 30 seconds, wherein the granules are controlled to be nearly spherical, so as to obtain wet enzyme pellet core granules with a diameter of 0.3-0.5mm, and preferably at a rotating speed of 60-80rpm;
Further, the wet enzyme pellet core particles are placed in a fluidized bed for drying, and the air inlet temperature is 60-70 ℃ and the drying is carried out under the vacuum for 0.3-0.5 time until the moisture is <5%, preferably the moisture is <4%, more preferably the moisture is <3%;
(3) The preparation method of the additive comprises the following steps:
weighing the auxiliary agent according to the weight ratio of the enzyme pill core to the auxiliary agent of 4:1, pouring the auxiliary agent into an oil bath pot, and heating to 90-110 ℃ to melt the auxiliary agent;
Pouring the dried enzyme pellet core into a fluidized bed, controlling the air inlet temperature of the fluidized bed to be 15-25 ℃ and the air quantity to be 60m < 3 >/h-170 m < 3 >/h, and spraying the auxiliary agent after the thermal dissolution on the enzyme pellet core by a peristaltic pump under the atomization pressure of 0.3-0.4Mpa to obtain the composite enzyme particles.
Further, in order to improve the hardness of the particles, a protective layer is sprayed outside the auxiliary agent layer according to the weight ratio of the composite enzyme particles to the protective layer of 9:1, wherein the protective layer comprises the following components in parts by weight of 20 parts of CMC, 30 parts of microcrystalline cellulose, 50 parts of calcium carbonate and 400 parts of water, and the components are mixed and emulsified into suspension.
Further, the spraying method of the protective layer comprises the following steps of controlling the air inlet temperature of a fluidized bed to be 40-60 ℃, the air quantity to be 60m < 3 >/h to 170m < 3 >/h, and the atomization pressure to be 0.5-0.7Mpa until all the protective layer is sprayed on enzyme particles.
The beneficial effects are that:
The invention aims to provide stable, environment-friendly and efficient composite enzyme granules and a preparation method thereof, wherein the composite enzyme granules are prepared by mixing powder with simple enzyme, so that the storage stability of the enzyme is improved, the retention rate of enzyme activity is improved by 20-30%, the mixing uniformity of the enzyme is improved by 20% after shaking with the same strength, the stress resistance in the use process is improved, the enzyme half life is prolonged by 30-50% after dissolution with high pH, and in addition, the dust and labor intensity in the use process and the influence of human factors on production are obviously reduced, the automation of industrial production is promoted, and the stability of the production process is improved to improve the product quality.
The specific embodiment is as follows:
In order to more clearly illustrate the advantageous features and properties of the present invention, a highly efficient bio-complex enzyme preparation granule and a method for preparing the same according to the present invention will be described in detail, and the following examples are only given for illustrating the enzyme preparation granule and the method for preparing the same, but are not intended to limit the scope of the present invention.
The invention aims to provide high-efficiency biological compound enzyme preparation particles and a preparation method thereof, so that the problems of dust hazard and mixing uniformity of powder compound enzyme in the using process are solved, and the storage and use stability of the enzyme in the compound enzyme are improved. The preparation method is safe, simple, convenient and efficient. The enzyme particles comprise an enzyme pellet core and an auxiliary agent, and the auxiliary agent can improve the action efficiency of the enzyme to a certain extent.
It is understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described in the following (embodiments) may be combined with each other to constitute a new or preferred technical combination scheme. Is limited to space and will not be described again.
Example 1A biological Complex enzyme preparation granule and method for preparing the same
The stable and efficient composite enzyme particle comprises an enzyme pellet core and an auxiliary agent layer, wherein the enzyme pellet core is used as a core part, the auxiliary agent layer is coated on the outer layer of the enzyme pellet core, and the enzyme pellet core comprises an enzyme component and a stabilizer;
The weight ratio of the enzyme pellet core to the auxiliary agent layer is 4:1;
The weight part ratio of the enzyme component to the stabilizer in the enzyme pill core is 8:2;
The enzyme component comprises, by weight, 3 parts of alkaline protease, 15 parts of alkaline pectase, 8 parts of alkaline lipase, 35 parts of neutral cellulase, 25 parts of xylanase, 5 parts of laccase, 6 parts of amylase and 3 parts of mannanase;
The stabilizer comprises the following components in parts by weight of 35 parts of sorbitol, 38 parts of dextrin, 21 parts of trehalose, 5.6 parts of calcium chloride and 0.4 part of copper sulfate;
The auxiliary agent layer comprises the following components in parts by weight of 60 parts of C12-C14 fatty alcohol polyoxyethylene ether, 1000 30 parts of polyethylene glycol and 10 parts of alkylphenol polyoxyethylene ether.
According to the composition and the dosage, the preparation method is as follows:
(1) Immobilization treatment of partial enzymes:
Embedding and immobilizing alkaline protease by sodium alginate and calcium chloride to improve stress resistance of the enzyme;
Dissolving alkaline protease in 2% sodium alginate solution, atomizing under 0.5Mpa, adding into 3% calcium chloride solution containing 0.2% glutaraldehyde, fixing for 0.5 hr, filtering, washing with distilled water for 3 times, and drying at low temperature.
The alkaline lipase is embedded with beta cyclodextrin to improve the catalytic activity and stress resistance;
And respectively weighing 10 times of the weight of the lipase, mixing and stirring uniformly, adding into a lipase phosphate buffer solution with the pH of 7-8, stirring, reacting for 1h, pouring the product into a beaker after the reaction is finished, washing with 65% ethanol solution, centrifuging, and drying the product at 45-50 ℃ to obtain the immobilized lipase.
Crosslinking and fixing alkaline pectinase by chitosan to improve alkali resistance and heat resistance;
Weighing alkaline pectase, adding into chitosan suspension, adding glutaraldehyde solution with concentration of 0.2% -0.3%, immobilizing for 4 hours at 25-30 ℃, filtering, washing, recovering precipitate, and drying at 45-50 ℃.
The alkali resistance of the amylase and laccase is improved by the montmorillonite adsorption immobilization treatment. The optimal immobilization condition is that montmorillonite with 5 times weight of enzyme is weighed and added into enzyme solution with pH of 6.5-8.0, the temperature is 15-20 ℃, and the adsorption immobilization time is 0.5-2 hours. Filtering or centrifuging the immobilized enzyme, recovering immobilized enzyme, oven drying at low temperature, pulverizing, and sieving;
(2) Preparing an enzyme pill core:
mixing all enzyme components and a stabilizer in the enzyme pellet core according to the weight ratio of 8:2, adding a microcrystalline cellulose disintegrating agent according to the weight ratio of 95:5 into the mixture according to the mixture, uniformly mixing, preparing a wet enzyme pellet core by adopting a wet rotary extrusion granulating method, and then drying the wet enzyme pellet core in a fluidized bed;
In the wet rotary extrusion granulating process, the proportion of water to materials (enzyme component, stabilizer and microcrystalline cellulose disintegrating agent) is controlled at 1.5:5, the temperature of the rotary extrusion materials is less than 50 ℃, a 30-mesh hole net is selected for extrusion granulating, the diameter of an extrusion strip is controlled to be 0.3-0.5mm, the extrusion strip is added into a shot blasting machine, the rotating speed is 80rpm, the shot blasting is carried out for 30 seconds, and the particles are controlled to be nearly spherical, so that wet enzyme pellet core particles with the diameter of 0.3-0.5mm are obtained;
drying the wet enzyme pellet core particles in a fluidized bed at 60-70 ℃ and 0.3-0.5 vacuum until the moisture is less than 3%;
(3) Spraying an auxiliary agent layer:
Weighing the auxiliary agent according to the weight ratio of the enzyme pill core to the auxiliary agent of 4:1, pouring the auxiliary agent into an oil bath pot, heating to 90-110 ℃, melting and spraying the mixture onto the surface of the enzyme pill core;
pouring the dried enzyme pellet core particles into a fluidized bed, controlling the air inlet temperature of the fluidized bed to be 15-25 ℃ and the air quantity to be 80
M3/h, spraying the auxiliary agent after the thermal dissolution on the enzyme pill core by a peristaltic pump under the atomization pressure of 0.3-0.4 Mpa.
(4) And (3) spraying a protective layer:
In order to improve the hardness of the granules, a protective layer is sprayed outside the auxiliary agent layer according to the weight ratio of enzyme granules to the protective layer of 9:1, and the protective layer comprises the following components in parts by weight of 20 parts of CMC, 30 parts of microcrystalline cellulose, 50 parts of calcium carbonate and 400 parts of water, and the components are mixed and emulsified into suspension.
The spraying method of the protective layer comprises the following steps of controlling the air inlet temperature of a fluidized bed to be 50 ℃, the air quantity to be 80m < 3 >/h and the atomization pressure to be 0.5Mpa until all the protective layer is sprayed on enzyme particles.
The particles prepared as described above are designated particle 1.
Example 2 Complex enzyme preparation particle and method for preparing the same
The composition and preparation method of the compound enzyme granule are the same as those of the enzyme of the example 1, except that no stabilizer is added. A complex enzyme pellet was prepared as described above and designated pellet 2.
Example 3 Complex enzyme preparation particle and method for preparing the same
The composition and preparation method of the compound enzyme granule are the same as those of the enzyme of the example 1, except that all enzymes are not immobilized. A complex enzyme pellet was prepared as described above and designated pellet 3.
Example 4 Complex enzyme preparation particle and method for preparing the same
The composition and preparation method of the compound enzyme granule are the same as those of the enzyme of the example 1, except that no auxiliary agent is sprayed. A complex enzyme pellet was prepared as described above and designated pellet 4.
Example 5A Complex enzyme preparation and method for preparing the same
Alkaline protease, alkaline pectase, alkaline lipase, neutral cellulase, xylanase, laccase, amylase and mannanase are weighed according to the composite granular enzyme components in the embodiment 1, and are uniformly mixed. And taking 5 enzyme powders at different positions, detecting enzyme activities, ensuring that the difference of detection values of the same enzyme activity at different positions is less than 2.0%, regarding the enzyme powders as uniform mixing, and ending uniform mixing operation.
Example 6 comparison of different Complex enzymes
The enzyme complex of examples 1 to 5 was stored in an incubator at 40℃and 30 to 40% humidity for 1 month, and the activity was measured, and the enzyme activity retention rate was calculated with the enzyme activity in the non-stored product being 100%.
Example 7 comparison of different Complex enzymes
The complex enzymes of examples 1-5 were weighed, placed in self-sealing bags of the same type, put in a shaker, shaken for 1 hour at 100 rpm, taken out, and the complex enzymes of the same mass were weighed from the same place to measure the activity, and the enzyme mixing uniformity was calculated with the enzyme activity in the product without shaking being 100%.
Example 8 comparison of different Complex enzymes
Weighing the compound enzyme of examples 1-5, adding into 9 times of buffer with pH of 9.0, detecting enzyme activity after dissolution, taking the enzyme activity as initial enzyme activity, incubating at 40 ℃, detecting solution activity during incubation, and calculating the time for enzyme activity to remain 50%.
Example 9 calculation
The enzyme activity detection criteria in this example are referred to as alkaline protease detection criteria GB/T23527.1, alkaline lipase activity detection criteria GB/T2583, neutral cellulase detection criteria QB/T2583, pectinase detection criteria QB/T4482, and in this example, a portion of the enzymes are selected as the evaluation subjects for the activity detection.
Enzyme activity retention = remaining enzyme activity after treatment ≡initial enzyme activity before treatment × 100%
Enzyme component mixing uniformity= (1- | enzyme activity after shaking-enzyme activity before shaking|/enzyme activity before shaking) ×100%
EXAMPLE 6 enzyme Activity survival
EXAMPLE 7 uniformity of mixing of enzyme Components
Example 8 half-life of enzyme component
In summary, compared with powder, the composite enzyme granule prepared by the invention has the advantages that the retention rate of enzyme activity is improved by 20-30%, the mixing uniformity of enzyme components is improved, the mixing uniformity of the enzyme components is improved and is unchanged after the enzyme components are shaken outside, the mixing uniformity of the composite enzyme powder is reduced by 20%, the stress resistance of the composite enzyme granule in the use process is improved, the composite enzyme granule is preserved under the same test condition, the half-life period of the enzyme is prolonged by 30-50%, and in addition, obviously, the dust and labor intensity in the use process and the influence of human factors on production are reduced, the automation of industrial production is promoted, and the stability of the production process is improved to improve the product quality.
Claims (8)
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