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Lysine and cellulase fermented and coated feed

A technology of cellulase and lysine, applied in fermentation, animal feed, animal feed, etc., can solve problems such as trouble, loss, easy mixing and unevenness

Active Publication Date: 2013-12-18
NINGXIA EPPEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this way, it is more troublesome to use, and it is easy to mix unevenly, and the corresponding utilization rate is reduced
[0006] The inventor imagines that in the process of preparing coated lysine products, cellulase and lysine fermentation broth are directly mixed, which can be mixed evenly and then coated, but it is found that it is limited by the coating process. The normal enzyme activity will be greatly weakened or even completely lost due to the high temperature and drying process

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Preparation of expression construct of heat-resistant and dehydration-resistant cellulase gene

[0059] According to the strain we found, we entrusted the Institute of Microbiology, Chinese Academy of Sciences to conduct sequence analysis, and found a new cellulase gene, the nucleotide sequence of which is shown in SEQ ID No: 1 of the sequence table, the encoded cellulase The amino acid sequence is shown in SEQ ID No:2. Then, according to the "Molecular Cloning Experiment Guide" and the operating guide of the commercial reagents used to construct the secretory expressed yeast, in short, the gene encoding the cellulase is used as a forward primer as shown in SEQ ID No: 3 of the sequence listing shown (the EcoR I endonuclease site was introduced), the reverse primer was shown in SEQ ID No: 4 of the sequence table (the Not I endonuclease site was introduced) and double-digested with EcoR I and Not I after amplification , and ligated with the pPIC3.5 plas...

Embodiment 2

[0061] Example 2 Fermentation preparation and activity determination of heat-resistant and dehydration-resistant cellulase

[0062] Inoculate the positive yeast transformant obtained in Example 1 into 50ml BMGY liquid medium (100mM potassium phosphate buffer (pH6.0), containing 1% yeast extract, 2% peptone, 1.34% yeast nitrogenous base, biological Prime 4*10 -5 %, glucose 2%, glycerol 1%), cultured at 30°C, 200rpm until OD600 reached 5.0.

[0063] Transfer the above culture to 1L BMGY liquid medium, cultivate at 30°C, 200rpm for 24 hours, add 5ml of methanol, and then continue to cultivate at 30°C, 200rpm for 6 days, during which 5ml of methanol was added every 24 hours, and the solution was controlled by ventilation. Oxygen (DO) is greater than 20%, and the pH is controlled at 6.0 (adjusted with ammonia water). After the cultivation is completed, filter with a 0.22 μm filter membrane and retain the supernatant, which is detected by SDS-PAGE to be rich in the protein corre...

Embodiment 3

[0067] Fermentative production of embodiment 3 L-lysine

[0068] As described in Chinese patent 201110065683.5, the pyridine nucleotide transhydrogenase variant gene (its nucleotide sequence is shown in SEQ ID No: 5 of the sequence listing) was cloned into the Escherichia coli-coryneform bacteria shuttle plasmid pMS2 ( Available from the American Type Microorganism Collection (ATCC), product number ATCC 67189) between EcoR I and Xba I endonuclease sites, electrotransformed into L-lysine fermented coryneform bacteria engineering bacteria (available from American Type Microorganisms Collection) Microorganisms Collection Center (ATCC), product number ATCC 31269), obtained coryneform bacteria engineering bacteria.

[0069] Insert the coryneform bacterium engineering bacterium into 20 cubic meters of fermenter with 0.5% inoculum (the culture medium formula is: 600 kilograms of glucose, 200 kilograms of sugarcane molasses, 520 kilograms of corn steep liquor, KH 2 PO 4 45...

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PUM

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Abstract

The invention relates to lysine and cellulase fermented and a coated feed, and provides a method for preparing an L-lysine product by fermentation. The method comprises the steps: fermenting to produce an L-lysine fermented liquor, mixing the L-lysine fermented liquor with high-temperature-resisting dewatered cellulase obtained through the fermentation, and coating by using fat after high temperature drying. In addition, the invention also provides a product produced by using the method, the high-temperature-resisting dewatered cellulase and the like.

Description

technical field [0001] The invention belongs to the field of fermentation of amino acids. Specifically, the invention relates to a method for fermenting and preparing L-lysine products, which includes fermenting and producing L-lysine fermentation broth, which can be dehydrated with fermentable high-temperature resistant The cellulase is mixed, dried at high temperature and coated with fat. In addition, the invention also provides the product produced by the method, the high temperature dehydration resistant cellulase and the like. Background technique [0002] Cellulase can effectively degrade cellulose-containing substances (such as feed), so adding cellulase to feed can enhance the absorption and utilization of feed, thereby saving feeding costs. For example, Chinese patent 96196760.9 discloses an enzyme supplement for ruminant feed, which contains cellulase. [0003] L-lysine is an important amino acid raw material, which can be used as condiment, food, feed additive,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K1/00A23K1/16C12P13/08C12N9/42
Inventor 马吉银陈崇安孟刚曹洪程耀东刘鑫
Owner NINGXIA EPPEN BIOTECH
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