A method for synergistically fermenting wheat straw to prepare high-quality biological feed

A technology for wheat straw and biological feed is applied in the field of synergistic degradation and fermentation of wheat straw into high-quality biological feed.

Active Publication Date: 2022-07-22
JIANGSU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Limited by the current situation of enzyme preparations on the market, the current bio-feed fermentation process pays more attention to the degradation of cellulose and hemicellulose, and there are few reports on the fermentation process using lignin as a breakthrough point, especially the synergistic fermentation preparation of bacteria and enzymes combined with small molecular substances Research on biological feed that meets the requirements of high protein content and low lignocellulose content is even rarer

Method used

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  • A method for synergistically fermenting wheat straw to prepare high-quality biological feed
  • A method for synergistically fermenting wheat straw to prepare high-quality biological feed
  • A method for synergistically fermenting wheat straw to prepare high-quality biological feed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The amino acid sequence of LiP (Protein ID: 1589173) in JGI database (https: / / jgi.doe.gov) Irpex lacteus CCBAS Fr.238 617 / 93v1.0 is shown as SEQ ID NO.3. Its coding sequence is codon optimized according to the codon preference of Schizosaccharomyces pombe, and the nucleotide sequence of the LiP gene after codon optimization is shown in SEQ NO.1.

[0037] Design primers to amplify the target gene sequence. The forward primer sequence is shown in SEQ ID NO.4, and the reverse primer sequence is shown in SEQ ID NO.5. 60°C for 30s, 72°C for 60s, 30 cycles). The primers were designed using the shuttle plasmid pREP as the template. The sequence of the forward primer is shown in SEQ ID NO.6, and the sequence of the reverse primer is shown in SEQ ID NO.7. , 72 ℃ 50s, 30 cycles). The target fragment was recovered and the product concentration was determined. figure 1 For the fragment map of the PCR product of the vector and the target gene, the above PCR product fragments were...

Embodiment 2

[0040] In this example, the enzymatic properties of recombinant LiP were determined.

[0041] The S. pombe-pREP-lip obtained in Example 1 was expanded and cultured in YES liquid medium, and the culture medium was collected. After the culture medium was concentrated by ultrafiltration tube, SDS-PAGE and Western Blot verified the extracellular secretion of target protein by Schizosaccharomyces pombe. figure 2 is the SDS-PAGE electropherogram of Schizosaccharomyces pombe exocrine protein, image 3 Western Blot of S. pombe exocrine protein; combined with figure 2 and image 3 It can be seen that the recombinant LiP is expressed extracellularly in Schizosaccharomyces pombe, and the apparent molecular weight is about 37kDa. The change of the amount of extracellular secreted protein with the culture time is as follows: Figure 4 As shown in the figure, it can be seen from the figure that the amount of extracellular secreted protein of S.pombe-pREP-lip reached the peak on the 6t...

Embodiment 3

[0046] In this example, the inoculation amount of S.pombe-pREP-lip and S.pombe-pREP-mnp is 2%, the inoculation amount of N.crassa is 2%, the inoculation amount of L.fermentium is 1%, and the inoculation amount of C.utilis is 1% %; the inoculation of the bacterial classification is all fermented seed liquid; the preparation method of the fermented seed liquid of the bacterial classification is as follows:

[0047] N.crassa: Take the frozen N.crassa, inoculate it on the slant of PDA solid medium, incubate at 30 °C for 3 days until the spores are abundant, filter with 8 layers of gauze in the ultra-clean workbench, and repeat with sterilized water. Rinse the mature spores of N. crassa on the slant of the PDA, and the eluate is the spore suspension, which is counted with a hemocytometer, and adjusted to the number of 1 × 10 spores. 9 N.crassa spore suspension was then added to the PDA culture medium at 1% inoculum, and cultivated at 120 rpm, 30° C., in a constant temperature shake...

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Abstract

The invention belongs to the field of biological feed, in particular to a method for synergistically degrading and fermenting wheat straw into high-quality biological feed. First, the lignin peroxidase gene from R. albicans was optimized and introduced into the defective Schizosaccharomyces pombe to obtain the lignin peroxidase-Schizophrenia pombe recombinant engineered strain; The strains were combined with Neurospora crassa, Lactobacillus fermentum, Candida rugugenens, and S.pombe‑pREP‑mnp fermentation strains producing manganese peroxidase, and added glucose, nitrogen source, laccase and glucose oxidase, and small Molecular substances are used to synergize bacteria and enzymes to ferment straw to obtain high-quality biological feed. The method rationally utilizes agricultural waste, promotes the conversion of lignocellulose in straw, increases the true protein content in the straw feed, degrades the lignocellulose content therein, significantly improves the quality and nutritional value of the fermented feed in a short time, and obtains high-quality straw fermented feed.

Description

technical field [0001] The invention belongs to the field of biological feed, in particular to a method for synergistically degrading and fermenting wheat straw into high-quality biological feed. Background technique [0002] With the annual increase of my country's grain output, the output of straw has also surged, and China produces about 900 million tons of straw every year. Due to the lack of scientific and rational application technology, these straws are mostly discarded as agricultural waste. With the continuous advancement of modern agriculture in my country, the rational and efficient utilization of straw resources has become the current research focus. my country is also a big country in animal husbandry, which produces a large amount of high-quality pasture and feed every year. At present, a large amount of high-quality pasture in China still relies on imports. Converting straw into high-quality biological feed can not only alleviate the current waste of straw r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/19C12N15/53C12N15/81A23K10/37A23K10/12A23K10/14C12R1/645
CPCC12N9/0065C12N15/815C12Y111/01014C12Y111/01013A23K10/37A23K10/12A23K10/14C12N2800/22A23V2400/143Y02P60/87
Inventor 陈华友梁晓玉蔡康涛李守志王振崔周磊王洪成朱道辰卜权
Owner JIANGSU UNIV
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