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Method for preparing high-purity lysine salt by fermentation and product thereof

A kind of technology of lysine sulfate and lysine, applied in the field of amino acid fermentation

Active Publication Date: 2015-09-30
NINGXIA EPPEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] After long-term research and experimentation by the inventor, it was unexpectedly found that adding a certain proportion of other types of amino acids to lysine sulfate with poor moisture absorption resistance , although it will reduce the purity of lysine sulfate in the product, it will significantly improve the moisture resistance of the lysine sulfate product, and the product of this purity will not affect its effectiveness as a lysine feed additive

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] The preparation of embodiment 1 RNA polymerase sigma-32 factor gene construct

[0075] According to the amino acid sequence of the protein accession number AAB18436.1 of NCBI (http: / / www.ncbi.nlm.nih.gov), the inventors designed moderate expression codons (non-expression-optimized codons), and obtained The pathway commissioned Shanghai Sangon Biotechnology Co., Ltd. to synthesize the gene encoding RNA polymerase sigma-32 factor and construct it into E. coli expression plasmid pET-20b(+) (available from Novagen, USA, product number Cat. No. 69739-3 )middle. The cloning process was carried out according to the "Molecular Cloning Experiment Guide" and the operation guide of the commercial reagents used. The brief process is as follows:

[0076] Synthesize nucleic acid fragments of the RNA polymerase sigma-32 factor gene by an automatic DNA synthesizer, phosphorylate the 5' ends of these nucleic acid fragments with T4 polynucleotide kinase (purchased from TaKaRa Co...

Embodiment 2

[0078] Example 2 Production of L-lysine by Escherichia coli fermentation

[0079] The L-lysine-producing Escherichia coli strain W3110(tyrA) / pCABD2 constructed by the method described in Chinese Patent No. 94194962 was transformed into a pET-sigma plasmid, and a positive clone was obtained after identification (named W3110(tyrA) / pCABD2 -sigma), the strains W3110(tyrA) / pCABD2 and W3110(tyrA) / pCABD2-sigma were respectively subjected to lysine fermentation experiments with reference to Chinese Patent No. 03120099. In short, the bacterial strain was inserted into the liquid LB medium for shaking culture until the OD500 reached 0.35, and was inserted into the lysine fermentation medium with a 5% inoculum size (the formula per liter of medium was: 100g glucose, 60g (NH 4 ) 2 SO 4 , 50g CaCO 3 , 35mL peptone-B (Soy Protein Enzymatic Hydrolysate, purchased from Shanghai Xinran Biotechnology Co., Ltd., total nitrogen content not less than 3%), 1g KH 2 PO 4 , 400mg MgSO 4 ·7H 2...

Embodiment 3

[0082] The refining of embodiment 3 L-lysine sulfate product

[0083] The supernatants obtained from strains W3110(tyrA) / pCABD2 and W3110(tyrA) / pCABD2-sigma described in Example 2 were respectively taken out by 1 / 4 volume and purified first: these supernatants were loaded on Sephadex G-10 The chromatographic column is eluted with pure water, and the fractions with a molecular weight of 100~200Da are collected, and the contents of L-lysine and other components in the fractions are quantified by HPLC, wherein: W3110(tyrA) / pCABD2 The composition distribution of the product is: lysine, 92.60%; threonine, 3.23%; serine, 1.70%; glutamic acid, 1.33; methionine, 0.76%; ); the composition distribution of W3110(tyrA) / pCABD2-sigma was: lysine, 80.73%; aspartic acid, 2.39%; threonine, 1.51%; serine, 0.89%; glutamic acid, 3.73%; valine Acid, 1.37%; Methionine, 0.68%; Isoleucine, 0.97%; Leucine, 1.97%; Tyrosine, 1.12%; Phenylalanine, 0.99%; Histidine, 1.50%; Arginine, 1.72%; other impur...

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Abstract

The invention discloses a method for preparing a high-purity lysine salt based on fermentation and a product prepared according to same, and provides a method for preparing L-lysine salt based on fermentation. According to the method, bacterium producing L-lysine is introduced with a gene which is regarded as no relevance with lysine metabolism, and the content of lysine in the obtained product is improved. In addition, the invention further provides a product prepared according to the method, the product can remarkably improve humidity resistance compared with the existing products containing L-lysine, and therefore is suitable for long-term storage; the preparation method is flexible, and the cost is controllable.

Description

technical field [0001] The present invention belongs to the field of amino acid fermentation. Specifically, the present invention relates to a method for fermenting and preparing products containing L-lysine sulfate, which includes innovatively introducing a lysine that is generally considered to be related to lysine into L-lysine-producing bacteria. Acid metabolism unrelated genes, and the lysine content in the obtained product can be improved. In addition, the invention also provides products and applications produced by the method. Background technique [0002] L-lysine is an important amino acid raw material, which can be used as condiment, food, feed additive, and also as an effective or auxiliary ingredient in health care products and medicines. It is widely used in food industry, feed industry, pharmaceutical industry and other chemical industries. In industry, especially lysine salts containing impurities (such as sulfate, hydrochloride), generally can be directly ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/08C12P13/06C12P13/10C12P13/12C12P13/14C12P13/20C12P13/22C12P13/24A23K1/16C12R1/19
Inventor 马吉银陈崇安孟刚
Owner NINGXIA EPPEN BIOTECH
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