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Gene for coding alanine isomerase and application of gene

An alanine and isomerase technology, applied in the fields of genetic engineering and enzyme engineering, can solve the problems of low yield, long process flow, easy environmental pollution, etc., and achieve the advantages of improving conversion efficiency, shortening process route and reducing environmental pollution. Effect

Inactive Publication Date: 2015-05-06
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] But so far, no matter what kind of chemical synthesis method is used to prepare D-alanine, there are disadvantages such as long process flow, many steps, complicated by-product removal process, low yield, high cost and easy pollution of the environment, thus limiting the production capacity of D-alanine. Production and marketing of alanine

Method used

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  • Gene for coding alanine isomerase and application of gene
  • Gene for coding alanine isomerase and application of gene
  • Gene for coding alanine isomerase and application of gene

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Experimental program
Comparison scheme
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Embodiment

[0035] 1. Library sources and plasmids

[0036] Yeast cDNA library and pPICZaA are preserved by our laboratory.

[0037] 2. Main reagents

[0038] The expression host strain P. pastoris X-33 (wild type) and the antibiotic Zeocin were purchased from Invitrogen. Both Pmll enzyme and XbaI enzyme were purchased from TaKaRa Bioengineering Company.

[0039] 3. Method

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PUM

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Abstract

The invention belongs to the field of genetic engineering and enzyme engineering, and particularly relates to a method for cloning and expressing a gene for coding alanine isomerase. The gene for coding the alanine isomerase is obtained from a constructed yeast cDNA library, and a nucleotide sequence of the gene is as shown in SEQ ID NO:1. After being cloned, the gene for coding alanine isomerase is connected with an expression plasmid to construct a recombinant expression plasmid; after being linearized and purified, the recombinant expression plasmid transforms a recipient bacterium, and is screened to obtain a genetically engineered bacterium; after being induced to express, the genetically engineered bacterium is purified and identified to obtain alanine isomerase. The gene disclosed by the invention adopts the alanine isomerase to realize conversion of L-alanine into D-alanine by a one-step enzyme method, so that the conversation efficiency is improved, the process route of the D-alanine is shortened, the production cost is reduced, and the environmental pollution is reduced.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and enzyme engineering, and specifically relates to a gene encoding alanine isomerase and application thereof. Background technique [0002] Alanine is one of the 20 basic amino acids that make up protein, an excellent transport tool for nitrogen in the blood, and an effective sugar-producing amino acid. D-alanine is contained in large organic tissues as a cell wall peptide muicylate or peptide antibiotic. D-alanine is a non-natural amino acid, an important organic chiral source, mainly used in the fields of chiral drugs, chiral additives, chiral auxiliary agents, etc. sex source. At present, D-alanine is mainly used in the production of new broad-spectrum antibiotics, D-alaninol, alanine protective agent in the process of peptide synthesis, food additives and cosmetics. Also D-alanine [0003] It is also widely used in functional edible sweeteners. D-alanine is an important raw material f...

Claims

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Application Information

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IPC IPC(8): C12N15/61C12N9/90C12N15/81C12N1/19C12R1/84
Inventor 黄时海李湘萍盘慧群黄广上鄢凯舟梁智群陈桂光苏辉兰王国盼李丽刘诗宇李连威付跃
Owner GUANGXI UNIV
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