L-histidine high-yielding strain and application thereof

A technology for producing histidine and bacteria, applied in the field of bioengineering, can solve problems such as late start of research

Inactive Publication Date: 2013-04-03
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Domestic research in this area started relatively late

Method used

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  • L-histidine high-yielding strain and application thereof

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1 starts the mensuration of bacterial strain initial output

[0026] The starting strain Serratia marcescens (Serratia marcescens) ATCC 31026 was activated by streaking, and the seed medium was prepared and transferred to the fermentation medium. The initial yield was measured to be 2.1g / L.

Embodiment 2

[0027] Example 2 Preparation of mutants resistant to 3-amino-1,2,4-triazole

[0028] The starting strain Serratia marcescens (Serratia marcescens) ATCC 31026 was mutagenized with diethyl sulfate at a concentration of 2% at 30°C for 20 minutes, and coated onto 3-amino-1,2,4-triazole ( 2mg / mL) resistance plate, cultivated at 30°C for 36h, and picked the strain grown on this medium as a mutant resistant to 3-amino-1,2,4-triazole. Carry out L-histidine fermentation test then, among these mutant strains, the mutant strain with the highest L-histidine production is called Serratia marcescens (Serratiamarcescens) A 07, and L-histidine production is 3.5g / L.

Embodiment 3

[0029] Example 3 Preparation of mutants resistant to 6-mercaptopurine

[0030]Serratia marcescens A 07 obtained in Example 2 was mutagenized with 250 μg / mL nitrosoguanidine at 30° C. for 20 minutes, and coated to 6-mercaptopurine (3 mg / mL) resistance Cultivate on a plate for 36 hours at 30° C., and pick the strain grown on this medium as a mutant resistant to 6-mercaptopurine. Carry out L-histidine fermentation test then, among these mutant strains, the mutant strain with the highest L-histidine production is called Serratia marcescens (Serratia marcescens) M 20, and the L-histidine production is 4.6g / L.

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Abstract

The invention provides an L-histidine high-yielding strain and an application thereof, and belongs to the field of a bioengineering technology. The invention provides Serratia marcescens ZJZG25 which is obtained by the following steps of: by taking Serratia marcescens ATCC (American Type Culture Collection) 31026 as a starting strain, carrying out diethyl sulfate (DES), nitrosoguanidine (NTG) and ultraviolet (UV) gradual-grade mutation; and adding analogues of L-histidine including 3-amino-1,2,4-triazole, 6-purinethol, histidine methyl ester, 2-thiouracil, D-histidine and the like into a basic culture medium and screening. The L-histidine is produced by a strain fermentation method; and compared with the starting strain, the capability of the accumulating high-level L-histidine is expressed. The ZJZ 625 is subjected to shake-flask culture for 72 hours and the yield of the L-histidine can reach to 9.7 g / L. The fermentation is a 3-L fermentation tank is carried out for 60 hours and the yield of the L-histidine can reach to 18.1 g / L.

Description

technical field [0001] An L-histidine high-yielding bacterium and its application belong to the technical field of bioengineering. Background technique [0002] L-histidine is an essential amino acid for infants and young children, and L-histidine can be synthesized in the body of adults, so it is a semi-essential amino acid for adults. L-histidine is mainly used in amino acid infusion and comprehensive amino acid reagents, heart circulation organ medicines and peptic ulcer medicine, treatment of anemia, rheumatic diseases, allergies, etc. L-histidine has a variety of physiological functions and is widely used in medicine, feed and food industries. At present, L-histidine is mainly extracted from pig blood meal hydrolyzate by ion exchange process in China. Due to high cost and high hydrolysis loss rate, it causes environmental pollution and is not suitable for industrial production. Therefore, the production of L-histidine by fermentation method is accelerated. imminent. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/24C12R1/43
Inventor 陈坚李江华侯颖堵国成
Owner JIANGNAN UNIV
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