CN113519692A - Fungus enzyme composition and application thereof in soybean hull fermentation - Google Patents

Fungus enzyme composition and application thereof in soybean hull fermentation Download PDF

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Publication number
CN113519692A
CN113519692A CN202110647547.0A CN202110647547A CN113519692A CN 113519692 A CN113519692 A CN 113519692A CN 202110647547 A CN202110647547 A CN 202110647547A CN 113519692 A CN113519692 A CN 113519692A
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soybean
fermentation
soybean hulls
enzyme composition
bacillus
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朱崇淼
杭苏琴
李英英
朱平华
马瑞雪
耿利利
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Nanjing Zhirun Biotechnology Co ltd
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Nanjing Zhirun Biotechnology Co ltd
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    • 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
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/14—Pretreatment of feeding-stuffs with enzymes
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • 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
    • 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)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
    • 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)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2477—Hemicellulases not provided in a preceding group
    • C12N9/248—Xylanases
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

Abstract

A bacterial enzyme composition and its application in soybean hull fermentation are disclosed, wherein the bacterial enzyme composition comprises cellulase, xylanase, lactobacillus, bacillus and yeast. Compared with physical methods and chemical methods, the technical method for fermenting the soybean hulls by combining the complex enzyme and the complex probiotics is simple, easy to operate, safe, environment-friendly and low in cost. Compared with a single enzymolysis method or single fermentation, the method can reduce the fiber in the soybean hulls, reduce the anti-nutritional factors such as antigen protein and urease in the soybean hulls, greatly improve the palatability and the nutritional value of the soybean hulls, effectively promote the development and the utilization of the soybean hulls in livestock and poultry feed, improve the utilization efficiency of the soybean hulls by animals, and has popularization value.

Description

Fungus enzyme composition and application thereof in soybean hull fermentation
Technical Field
The invention relates to the technical field of agricultural biology, in particular to a fungal enzyme composition and application thereof in soybean hull fermentation.
Background
The soybean demand of China is more than 1 hundred million tons every year, more than 80 percent of soybeans need to be imported, the soybean hulls are used as byproducts of soybean processing, and the annual output of China reaches 1200 million tons. Soybean hulls, which have a high crude fiber content (about 34%) but a low lignin content (< 2%), are a potential feed resource for animals due to their high fiber, high energy and price advantages. The soybean hulls are added into the daily ration of the livestock and poultry or part of coarse feed is replaced, so that the feeding ingredients can be reduced, the shortage of feed resources can be relieved, and the growth (production) performance of animals can be improved. However, due to the existence of high content of cellulose, hemicellulose and various anti-nutritional factors, the addition of excessive soybean hulls can cause damage to animal bodies, hinder the digestion and absorption of nutrients by the bodies, and limit the utilization of the soybean hulls as feed resources. At present, common treatment methods for soybean hulls include a physical method, a chemical method, an enzyme preparation treatment method and a microbial fermentation method, so that anti-nutritional ingredients in the soybean hulls can be eliminated, and the nutritional value of feed is improved.
The physical method mainly comprises an extrusion expansion method, a steam explosion method, a heat treatment method and the like. The physical method can eliminate the fiber and anti-nutritional factors in the soybean hull, but the physical methodThe cost for processing the soybean hulls is high, and the feasibility in production is low. The chemical method is mainly characterized in that chemical reagents are added into raw materials, and common chemical reagents comprise sulfuric acid, hydrochloric acid, sodium hydroxide (NaOH) and calcium hydroxide (Ca)2OH) and ethanol. Although the treatment with the chemical agent can effectively degrade the fiber and the anti-nutritional factors, the chemical agent remained in the substrate may damage the health of the animal body and is inconvenient to operate, so that the feed cost is increased. The enzyme preparation treatment and the microbial fermentation are widely concerned with the advantages of simple operation, obvious effect, low cost and the like. The enzyme preparation can degrade indigestible carbohydrate in the soybean hulls into small molecular substances which are easily absorbed by the body, but bitter substances generated in the enzymolysis process can influence the palatability of the feed. The enzyme produced by microbial fermentation alone cannot completely meet the amount of enzyme required for degrading feed raw materials, the quality of a fermentation product is determined by the activity and growth condition of the microbes in the fermentation process, the fermentation process is not easy to control, and the feed quality is unstable.
Disclosure of Invention
The invention provides a fungal enzyme composition and application thereof in soybean hull fermentation, which can improve the nutritive value of soybean hulls, promote the utilization of the soybean hulls in livestock and poultry feed and improve the utilization rate of the soybean hulls. Cellulose, xylanase, lactobacillus, bacillus and yeast are fermented and combined to degrade anti-nutritional ingredients such as fiber, urease and antigen protein in the soybean hulls, and the nutritional value of the soybean hulls is improved.
The technical scheme is that the bacterial enzyme composition comprises cellulase, xylanase, lactobacillus, bacillus and saccharomycetes.
The Lactobacillus is Lactobacillus plantarum ZR1(Lactobacillus plantarum ZR1), the Bacillus is Bacillus natto ND1(Bacillus natto ND1), and the yeast is Saccharomyces cerevisiae ZR 4.
The application of the fungal enzyme composition in soybean hull fermentation.
The application comprises the specific steps of uniformly mixing soybean hull, bran and urea, adding into a fermentation bag, wherein the mass ratio of the soybean hull to the bran to the urea is 100:10:1, and adding water for uniformly mixing; respectively inoculating lactobacillus plantarum ZR1, bacillus natto ND1 and saccharomyces cerevisiae ZR4 to a fermentation bag in a unit of mL/g in an inoculation amount of 1 wt.%; adding cellulase and xylanase into fermentation bags respectively in the amount of 100U/g soybean hull and 250U/g soybean hull, mixing, and fermenting in a 37 deg.C incubator for 5 d.
The effective viable count of the Lactobacillus plantarum ZR1, the Bacillus natto ND1 and the Saccharomyces cerevisiae ZR4 is respectively 108CFU/mL。
Compared with physical methods and chemical methods, the method has the advantages that the technical method for fermenting the soybean hulls by combining the complex enzyme and the complex probiotics is simple, easy to operate, safe, environment-friendly and low in cost. Compared with a single enzymolysis method or single fermentation, the method can reduce the fiber in the soybean hulls, reduce the anti-nutritional factors such as antigen protein and urease in the soybean hulls, greatly improve the palatability and the nutritional value of the soybean hulls, effectively promote the development and the utilization of the soybean hulls in livestock and poultry feed, improve the utilization efficiency of the soybean hulls by animals, and has popularization value.
Drawings
FIG. 1 is a schematic diagram showing the determination of the addition amounts of cellulase (A) and xylanase (B);
FIG. 2 is a schematic diagram showing the effect of complex enzyme addition on soybean hull reducing sugar (A), crude protein (B), neutral detergent fiber (C) and acidic detergent fiber (D);
FIG. 3 is a schematic diagram showing the effect of bacterial enzyme-binding fermentation on soybean hull lactic acid (A), pH (B) and viable count (D-F); wherein, the microorganisms comprise non-enzymatic Enzymes, added Enzymes, non-added Bacteria and added Bacteria; note: *: significant difference between the groups with and without addition, #: the difference between the enzyme-added and enzyme-not-added groups was significant.
Detailed Description
The following examples are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.
Example 1 determination of the amount of fiber degrading enzyme added
1. Raw materials and sources
The soybean hull, the bran and the urea are all provided by the Qinghe coarse cereal cooperative, Nantong, Jiangsu province, and are crushed and screened by 40 meshes for later use.
2. Species of microorganism
Lactobacillus plantarum ZR1(Lactobacillus plantarum ZR1) and Saccharomyces cerevisiae ZR4(Saccharomyces cerevisiae ZR4) were both purchased from the China general microbiological culture Collection center of the Committee for culture Collection of microorganisms; bacillus natto ND1(Bacillus natto ND1) was isolated from Yijia probiotic biological feed starter (Yijia probiotic engineering Co., Ltd., Henan, Zheng Zhou) and the growth characteristics of 3 strains of bacteria were determined.
3. Screening of enzyme addition amount
Adding soybean hulls into a fermentation bottle, adding water, mixing uniformly, adding cellulase into the fermentation bottle in an amount of 0, 100, 500 and 1000U/g (xylanase is 0, 100, 250 and 500U/g) respectively, placing the fermentation bottle in a 37 ℃ constant temperature incubator for 48h of enzymolysis, and placing the fermentation bottle in a 90 ℃ constant temperature water bath kettle for 10min after the enzymolysis is finished to terminate the enzymolysis reaction. After enzymolysis, the amount of reducing sugar in the soybean hulls is detected, and the optimal addition amounts of cellulase and xylanase are determined to be 100U/g and 250U/g respectively. And a compound enzyme is prepared according to the addition amount of the two enzymes and added into the soybean hulls, and compared with the effect of a single enzyme, the effect of the compound enzyme is superior to that of the single enzyme.
Example 2 specific steps of fungal enzyme in combination with Soybean Peel fermentation technology
1. Uniformly mixing 200g of soybean hull, 20g of bran and 2g of urea, adding into a fermentation bag, adding water, and uniformly mixing;
2. inoculating the mixed strain with the inoculum (Lactobacillus plantarum ZR1, Bacillus natto ND1 and Saccharomyces cerevisiae ZR4, with effective viable count of 108CFU/mL) were inoculated to the fermentation bags at 1% (v/w) inoculum size, respectively;
3. adding cellulase and xylanase into a fermentation bag according to the amount of 100U/g rapeseed meal and 250U/g soybean hull respectively, uniformly mixing, and transferring the fermentation bag into a 37 ℃ incubator for fermentation for 5 d.
Example 3 application of fungal enzyme in combination with fermented Soybean Peel technology
Effect of fungal enzyme binding on Soybean Peel nutritive value
The effect of the complex enzyme and the complex bacteria in combination with the fermented rapeseed meal was studied, and the test was divided into 4 groups (control group, enzymolysis group, fermentation group, and enzymolysis + fermentation group) and subjected to enzymolysis fermentation according to the procedure of example 2. As a result, the number of viable bacteria and lactic acid content in the enzymolysis and fermentation group were significantly increased and pH was significantly decreased (fig. 3) compared to the other three groups, and the content of fiber was significantly decreased in the enzymolysis and fermentation group compared to the non-enzymolysis group (control group, fermentation group) (table 1). The fiber degrading enzyme enzymolysis promotes the fermentation of the compound bacteria, and simultaneously degrades the anti-nutritional components such as antigen protein, urease, fiber and the like in the soybean hull.
The soybean hull has the fiber content of about 34 percent and contains a large amount of anti-nutritional factors, and the application of the soybean hull in the feed is limited due to the existence of the fiber and the anti-nutritional factors.
Figure BDA0003107823120000041

Claims (5)

1. A bacterial enzyme composition is characterized by consisting of cellulase, xylanase, lactobacillus, bacillus and yeast.
2. The bacterial enzyme composition according to claim 1, wherein the lactobacillus is lactobacillus plantarum ZR1 (ZR 1: (ZR C)Lactobacillusplantarum ZR1), the bacillus is natto bacillus ND1(Bacillus natto)Bacillusnatto ND1, the yeast is Saccharomyces cerevisiae (Saccharomyces cerevisiae)Saccharomyces cerevisiae ZR4）。
3. Use of the fungal enzyme composition of claim 1 or 2 in the fermentation of soybean hulls.
4. The application of the soybean milk powder is characterized in that the soybean milk powder, the bran and the urea are uniformly mixed and added into a fermentation bag, wherein the mass ratio of the soybean milk powder to the bran to the urea is 100:10:1, and water is added for uniform mixing; respectively inoculating lactobacillus plantarum ZR1, bacillus natto ND1 and saccharomyces cerevisiae ZR4 to a fermentation bag in a unit of mL/g in an inoculation amount of 1 wt.%; adding cellulase and xylanase into fermentation bags respectively in the amount of 100U/g soybean hull and 250U/g soybean hull, mixing, and fermenting in a 37 deg.C incubator for 5 d.
5. The use according to claim 4, wherein the effective viable count of Lactobacillus plantarum ZR1, Bacillus natto ND1 and Saccharomyces cerevisiae ZR4 is 10 respectively8 CFU/mL。
CN202110647547.0A 2021-06-09 2021-06-09 Fungus enzyme composition and application thereof in soybean hull fermentation Pending CN113519692A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114767757A (en) * 2022-04-14 2022-07-22 江西省科学院生物资源研究所 Accurate fermentation method of compound Chinese herbal medicine by-product and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109170138A (en) * 2018-10-09 2019-01-11 江门市澳保生物科技有限公司 A kind of technique that bacterium-enzyme cooperative fermentation produces high small peptide fermented bean dregs
CN110074252A (en) * 2019-05-20 2019-08-02 浙江康星生物科技有限公司 A kind of the fermented feed processing method and application of the high efficiency, low cost based on bean dregs

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109170138A (en) * 2018-10-09 2019-01-11 江门市澳保生物科技有限公司 A kind of technique that bacterium-enzyme cooperative fermentation produces high small peptide fermented bean dregs
CN110074252A (en) * 2019-05-20 2019-08-02 浙江康星生物科技有限公司 A kind of the fermented feed processing method and application of the high efficiency, low cost based on bean dregs

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114767757A (en) * 2022-04-14 2022-07-22 江西省科学院生物资源研究所 Accurate fermentation method of compound Chinese herbal medicine by-product and application thereof

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