阅读说明：本技术 一种秸秆混合发酵饲料及其生产方法 (Straw mixed fermentation feed and production method thereof ) 是由 江正兵 宋慧婷 李华南 肖文静 于 2019-09-25 设计创作，主要内容包括：本发明涉及生物发酵领域,尤其涉及一种秸秆混合发酵饲料及其生产方法。秸秆混合发酵饲料的生产方法,包括步骤：(1)配制混合发酵制剂：将全细胞纤维素酶和乳酸菌制剂溶于水中制得混合发酵液,所述全细胞纤维素酶为分泌纤维素酶的重组酿酒酵母菌；(2)将混合发酵液与秸秆混合均匀,混合均匀后装袋密封进行青贮发酵。本发明的秸秆混合发酵饲料能够提高秸秆饲料中纤维素、半纤维素的降解效率,而且能够提高秸秆饲料中单细胞蛋白含量,提升秸秆饲料品质。(The invention relates to the field of biological fermentation, in particular to a straw mixed fermentation feed and a production method thereof. The production method of the straw mixed fermented feed comprises the following steps: (1) preparing a mixed fermentation preparation: dissolving whole-cell cellulase and a lactobacillus preparation in water to prepare a mixed fermentation liquid, wherein the whole-cell cellulase is recombinant saccharomyces cerevisiae for secreting cellulase; (2) and uniformly mixing the mixed fermentation liquor and the straws, bagging and sealing after uniform mixing for ensiling fermentation. The straw mixed fermentation feed can improve the degradation efficiency of cellulose and hemicellulose in the straw feed, can improve the content of single cell protein in the straw feed, and can improve the quality of the straw feed.)

1. A production method of straw mixed fermentation feed is characterized by comprising the following steps:

(1) Preparing mixed fermentation preparation

dissolving whole-cell cellulase and a lactobacillus preparation in water to prepare a mixed fermentation liquid, wherein the whole-cell cellulase is recombinant saccharomyces cerevisiae for secreting cellulase;

(2) And uniformly mixing the mixed fermentation liquor and the straws, bagging and sealing after uniform mixing for ensiling fermentation.

2. the production method of the straw mixed fermentation feed as claimed in claim 1, wherein the mass ratio of the mixed fermentation liquid to the straw is 1-2: 1-3; the concentration of the whole-cell cellulase in the mixed fermentation liquid is 40-60 g/L, and the concentration of the lactobacillus preparation is 10-20 g/L.

3. The method for producing straw mixed fermentation feed according to claim 1, wherein the method for preparing the whole-cell cellulase comprises the following steps:

(1) Construction of cellulase expression vector pHBM-368-PGK-CBH2

inserting a target gene CBH2 with a nucleotide sequence shown in SEQ ID No.1 into a vector pHBM368-PGK by using a genetic engineering technology to obtain a cellulase expression vector pHBM-368-PGK-CBH 2;

(2) Transforming a cellulase expression vector pHBM-368-PGK-CBH2 into saccharomyces cerevisiae, and screening through an auxotroph culture medium to obtain whole-cell saccharomyces cerevisiae cellulase;

(3) the whole-cell saccharomyces cerevisiae cellulase is used as an original preparation strain, and whole-cell cellulase particles are obtained through the processes of inoculation activation, shaking culture, enlarged transfer, fermentation tank culture and low-temperature plasmid drying.

4. The method for producing straw mixed fermented feed according to claim 1, wherein the mixed fermentation broth is uniformly sprayed on the straw in the step (2), and the straw is repeatedly stirred during the spraying process, and then bagged, sealed and subjected to ensiling fermentation.

5. The production method of the straw mixed fermented feed as claimed in claim 1, wherein the straw is corn straw cut to a length of 1-3 cm.

6. The method for producing straw mixed fermented feed according to claim 1, wherein the lactobacillus preparation is a bio-storage modifier.

7. The straw mixed fermented feed is characterized by comprising a base material and a mixed fermented bacterial liquid; the base material comprises straws, and the mixed zymocyte liquid is prepared from whole-cell cellulase, a lactobacillus preparation and water; the whole-cell cellulase is recombinant saccharomyces cerevisiae secreting cellulase.

8. The straw mixed fermented feed as claimed in claim 7, wherein the mass ratio of the mixed fermentation liquid to the base material is 1-2: 1-3; the concentration of the whole-cell cellulase in the mixed fermentation liquid is 40-60 g/L, and the concentration of the lactobacillus preparation is 10-20 g/L.

Technical Field

The invention relates to the field of biological fermentation, in particular to a straw mixed fermentation feed and a production method thereof.

Background

The straw resources in China are rich, and it is estimated that 3.27 million tons of corn straws are produced in China every year. Most of the corn straws are naturally decomposed or directly burned, which can not only destroy organic matters on the surface layer of the soil, but also cause air pollution and harm human health. The corn straw is rich in organic matters, the protein and the fat also occupy a certain proportion, the basic organization mainly comprises cellulose, medium cellulose and lignin, and the corn straw has high development and utilization values in the aspects of feed, fertilizer, fuel edible fungus aggregate, processing raw materials and the like. The straw feed is one of effective ways for solving the problems of excess straw resources and unreasonable utilization, at present, the main treatment methods of the feed are divided into a chemical treatment method, a physical treatment method and a biological treatment method, and the biological treatment method is widely concerned due to the characteristics of low energy consumption, small environmental pollution, simple and convenient operation, no danger and the like.

At present, the biological treatment method for straw feed mainly depends on microorganisms to carry out a silage fermentation process to realize biodegradation and biological conversion of relevant components in straws, silage is a process for obtaining feed by carrying out anaerobic fermentation on the straws through lactic acid bacteria, and the key factor for ensuring successful silage is to keep lactic acid bacteria to continuously accumulate lactic acid and maintain a lower pH value, but sufficient soluble sugar supply is required for propagation of the lactic acid bacteria, and the continuity of the silage process cannot be maintained because the soluble carbohydrates in the straws are less.

Disclosure of Invention

In order to solve the problems, the invention aims to provide a straw mixed fermentation feed and a production method thereof, which can improve the degradation efficiency of cellulose and hemicellulose in the straw feed, improve the content of single-cell protein in the straw feed and improve the quality of the straw feed.

in order to achieve the purpose, the invention designs a production method of straw mixed fermentation feed, which is characterized by comprising the following steps:

(1) Preparing mixed fermentation preparation

dissolving whole-cell cellulase and a lactobacillus preparation in water to prepare a mixed fermentation liquid, wherein the whole-cell cellulase is recombinant saccharomyces cerevisiae for secreting cellulase;

(2) And uniformly mixing the mixed fermentation liquor and the straws, bagging and sealing after uniform mixing for ensiling fermentation.

Compared with the traditional silage method, the method has the advantages that the whole-cell cellulase is added, the whole-cell cellulase is the recombinant saccharomyces cerevisiae, the recombinant saccharomyces cerevisiae secretes the cellulase, the whole-cell cellulase taking the whole cell as a carrier is combined with lactic acid bacteria to carry out the forage treatment of the straws to generate a synergistic effect, the whole-cell cellulase degrades the cellulose in the straws, degradation products provide nutrient components required by fermentation for the lactic acid bacteria, the fermentation process of the lactic acid bacteria is maintained, the treatment effect of the corn straws is improved, and meanwhile, the activity of the lactic acid bacteria is utilized to remove the products decomposed by the cellulase and remove the feedback inhibition effect of the cellulase; and the whole-cell saccharomyces cerevisiae cellulase is used for self-propagating in the fermented corn straws, so that the fermentation of the lactic acid bacteria can be promoted, the restriction of small addition amount can be avoided, the cost of the cellulase is reduced, and the saccharomyces cerevisiae can provide a large amount of rich single-cell protein and can improve the feeding quality of the corn straws.

As a preferred scheme, the mass ratio of the mixed fermentation liquor to the straws is 1-2: 1-3; the concentration of the whole-cell cellulase in the mixed fermentation liquid is 40-60 g/L, and the concentration of the lactobacillus preparation is 10-20 g/L.

Preferably, the preparation method of the whole-cell cellulase comprises the following steps:

(1) Construction of cellulase expression vector pHBM-368-PGK-CBH2

Inserting a target gene CBH2 with a nucleotide sequence shown in SEQ ID No.1 into a vector pHBM368-PGK by using a genetic engineering technology to obtain a cellulase expression vector pHBM-368-PGK-CBH 2;

(2) transforming a cellulase expression vector pHBM-368-PGK-CBH2 into saccharomyces cerevisiae, and screening through an auxotroph culture medium to obtain whole-cell saccharomyces cerevisiae cellulase;

(3) The whole-cell saccharomyces cerevisiae cellulase is used as an original preparation strain, and the whole-cell cellulase is obtained through the processes of inoculation activation, shaking culture, enlarged transfer, fermentation tank culture and low-temperature plasmid drying.

Preferably, in the step (2), the mixed fermentation liquid is uniformly sprayed on the straws, the straws are repeatedly stirred in the spraying process, and then the straws are bagged, sealed and subjected to ensiling fermentation.

As a preferred scheme, the straws are corn straws which are cut to be 1-3 cm in length.

Preferably, the lactic acid bacteria preparation is a biological storage material modulator.

Drawings

FIG. 1 is a schematic diagram showing the gene recombination process of cellulase expression vector pHBM-368-PGK-CBH 2;

FIG. 2 is a graph showing the change in whole cell cellulase concentration during the preparation process;

FIG. 3 is a diagram showing the change of enzyme activity of cellulase in the preparation process.

Detailed Description

For a better understanding of the present invention, reference will now be made in detail to the present invention, examples of which are illustrated in the accompanying drawings.