Method for preparing high purity lysine sulfate through fermenting

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

Active Publication Date: 2013-02-13
NINGXIA EPPEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Through the inventor's long-term research and experiments, it has been found surprisingly that adding a certain proportion of other types of amino acids in the lysine sulfate with poor hygroscopicity resistance will reduce the purity of the lysine sulfate in the product, but It will significantly improve the moisture absorption resistance of lysine sulfate products, and the purity of the product will not affect its effectiveness as a lysine feed additive

Method used

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  • Method for preparing high purity lysine sulfate through fermenting
  • Method for preparing high purity lysine sulfate through fermenting
  • Method for preparing high purity lysine sulfate through fermenting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] The preparation of embodiment 1 RNA polymerase sigma-32 factor gene construct

[0076] According to the amino acid sequence of the protein accession number AAB18436.1 of NCBI (http: / / www.ncbi.nlm.nih.gov), the inventors designed a moderate expression codon (non-expression optimized codon), 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:

[0077] Synthesize nucleic acid fragments of the RNA polymerase sigma-32 factor gene by an automatic DNA synthesizer, use T4 polynucleotide kinase (purchased from TaKaRa company) to phosphorylate the 5' ends of these nucleic acid fragments, ...

Embodiment 2

[0079] Embodiment 2 Escherichia coli fermentation produces L-lysine

[0080] 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 as W3110(tyrA) / pCABD2 -sigma), the strains W3110(tyrA) / pCABD2 and W3110(tyrA) / pCABD2-sigma were 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 formulation of the medium per liter 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., the total nitrogen content is not less than 3%), 1g KH 2 PO 4 , 400mgMgSO 4 ·7H 2 O,...

Embodiment 3

[0084] The refining of embodiment 3L-lysine sulfate product

[0085] 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 Chromatographic column, eluted with pure water, collected fractions with a molecular weight between 100 and 200Da, and quantified the L-lysine content and other components in the fractions by HPLC, wherein: W3110(tyrA) / pCABD2 The composition distribution of lysine is as follows: 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 impurity components...

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Abstract

The invention provides a method for preparing high purity lysine sulfate through fermenting. The method includes that a gene irrelative to lysine metabolism is led into bacteria producing L-lysine, and lysine content in the obtained product can be increased. In addition, the invention further provides products produced by the method. Compared with the existing products containing L-lysine, the products can remarkably improve hygroscopicity resistance and are suitable for being stored for a long time. The preparation method is good in flexibility and controllable in cost.

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 u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/04C12P13/08A23K1/16
Inventor 马吉银陈崇安孟刚
Owner NINGXIA EPPEN BIOTECH
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