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Fermentation and coating of lysine and xylanase

A technology of xylanase and lysine, which is applied in the field of fermentation to prepare L-lysine products, and can solve problems such as low utilization rate, loss, and weakened enzyme activity

Active Publication Date: 2013-10-16
NINGXIA EPPEN BIOTECH +1
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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 low
[0006] The inventor imagines that xylanase and / or cellulase are directly mixed with lysine fermentation broth in the process of preparing coated lysine products, which can be mixed evenly and then coated, however However, it is found that due to the high temperature and drying process to be experienced in the coating, the conventional enzyme activity will be greatly weakened or even completely lost

Method used

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  • Fermentation and coating of lysine and xylanase
  • Fermentation and coating of lysine and xylanase
  • Fermentation and coating of lysine and xylanase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1 The preparation of the expression construct of the heat-resistant and dehydration-resistant xylanase gene

[0057] According to the strain we found, we commissioned the Institute of Microbiology, Chinese Academy of Sciences to conduct sequence analysis, and found a new xylanase gene, whose nucleotide sequence is shown in SEQ ID No: 1 in the sequence table, and the encoded xylan The amino acid sequence of the enzyme 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 xylanase is used as a forward primer such as SEQ ID No: 3 of the sequence listing Shown (introduced EcoR I endonuclease site), reverse primer as shown in the SEQ ID No of sequence listing: 4 (introduced Not I endonuclease site) amplified with EcoR I and Not I double enzyme and ligated with the pPIC3.5 plasmid (purchased from Invitro...

Embodiment 2

[0058] Example 2 Fermentation preparation and activity determination of heat-resistant and dehydration-resistant xylanase

[0059] 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.

[0060] 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 8 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 to be 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 proteins correspo...

Embodiment 3

[0064] The fermentative production of embodiment 3L-lysine

[0065] 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 ( Can be purchased from the American Type Microorganisms Collection (ATCC, product number ATCC 67189) between the EcoR I and Xba I endonuclease sites, electrotransformed into L-lysine fermented coryneform bacteria engineering bacteria (available from the American Type Microorganisms Collection) Microorganisms Collection Center (ATCC), product number ATCC 31269), obtained coryneform bacteria engineering bacteria.

[0066] Insert this coryneform bacterium engineering bacterium with 0.5% inoculation amount into 20 cubic meters of fermenters (the culture medium formula is: 600 kilograms of glucose, 200 kilograms of sugarcane molasses, 520 kilograms of corn steep li...

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Abstract

The present invention provides a fermentation method for preparing an L-lysine product. The method comprises steps of: producing an L-lysine fermentation broth through fermentation; mixing the fermentation broth with high temperature dehydration resistant xylanase which can be obtained by fermentation; and drying the mixture at high temperature, and coating with fat. In addition, the present invention also provides a product produced by the method and high temperature dehydration resistant xylanase, etc.

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 xylanase is mixed, dried at high temperature and coated with fat. In addition, the present invention also provides the product produced by the method and the high-temperature-resistant dehydration xylanase and the like. Background technique [0002] Xylanase can effectively degrade lignin-containing substances (such as feed), so adding xylanase 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 xylanase. [0003] L-lysine is an important amino acid raw material, which can be used as condiment, food, feed additiv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/24A23K1/16A23K1/165C12P13/08C12N15/56C12N15/63C12N1/19C12R1/15C12R1/84
Inventor 马吉银陈崇安孟刚曹洪程耀东刘鑫
Owner NINGXIA EPPEN BIOTECH
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