A kind of method of producing L-leucine by fermentation

A leucine and fermentation end technology, applied in the field of fermentation to produce L-leucine, can solve the problems of complex process and high cost, and achieve the effects of simple and easy control of the process, low cost, and shortened fermentation time

Active Publication Date: 2018-04-27
HENAN JULONG BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzyme used in the enzymatic production of L-leucine needs to be produced by microbial fermentation and extracted and refined. The process is relatively complicated and the cost is high, so it has not been widely used at present.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for producing L-leucine by fermentation, comprising the following steps:

[0023] Put 600ml of LB medium into a 2L shake flask, and insert the slant bacteria (brevibacterium flavum TL0412 (Met - +2-TA r +-α-AB r +β-HL r +SG r +Rif r , Tianjin University of Science and Technology Metabolic Control Fermentation Laboratory preservation strain), then put the shaker flask on the shaker at a stirring speed of 110r / min and a temperature of 31°C for 28 hours to obtain a seed solution for use; then inoculate at 10% Put the seed liquid into a 10L fermenter with a fermentation medium, and carry out fermentation culture at a temperature of 31°C; the fermentation medium includes the following carbon-containing compounds and nitrogen-containing compounds in mass percentages: anhydrous Glucose 2%, corn steep liquor 6%, ammonium sulfate 1.5%, ammonium acetate 1.0%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.2%, methionine 0.04%, isoleucine 0.006% and glutami...

Embodiment 2

[0027] A method for producing L-leucine by fermentation, comprising the following steps:

[0028] Put 600ml of LB medium into a 2L shake flask, insert the slant strain, then place the shake flask on a shaker at a stirring speed of 110r / min and a temperature of 31°C, and cultivate for 28h to obtain a seed solution for later use; Then, according to 10% inoculum size, the seed solution was inserted into a 10L fermentor with fermentation medium, and the temperature was 32°C for fermentation; the fermentation medium included the following carbon-containing compounds and Nitrogen compounds: anhydrous glucose 2%, corn steep liquor 6%, ammonium sulfate 1.5%, ammonium acetate 1.0%, potassium dihydrogen phosphate-0.1%, magnesium sulfate 0.2%, methionine 0.04%, isoleucine 0.006% and glutamine acid 0.07%; the amount of trace elements and vitamins in the fermentation medium is VB 1 300ug / L, VH50ug / L, FeSO 4 0.05mg / L, MnSO 4 0.03mg / L and ZnSO 4 0.05mg / L, the mass percentage of the ad...

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PUM

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Abstract

The invention discloses a method for fermenting and producing L-leucine, which comprises the following steps: loading LB medium into a shake flask, inserting slant strains, and then placing the shake flask on a shaker for 28 hours of cultivation to obtain seed liquid, Standby; then the seed liquid is connected to a fermenter with a fermentation medium for fermentation and cultivation; during the fermentation process, by controlling the stirring speed and air volume, the dissolved oxygen in the first 20 hours of the fermentation process is 6-8 mg / L, and the dissolved oxygen after 20 hours is 6-8 mg / L. 3-6mg / L; the pH of the fermentation process is controlled by automatic feeding of ammonia water, and the pH is controlled to be 6.4-6.7 before the OD value of the bacteria reaches 25. After the OD value of the bacteria increases to 25, the pH increases by 0.1 per hour until it increases Remains unchanged after 7.2; L-leucine is extracted and refined after fermentation. The method of the invention shortens the fermentation period, improves the fermentation output, has simple and easy control technology, and is low in cost, and is very suitable for large-scale industrial application.

Description

technical field [0001] The invention belongs to the technical field of amino acid production, and mainly relates to a method for producing L-leucine by fermentation. Background technique [0002] L-leucine, also known as leucine, was first isolated from cheese by Proust in 1819, and later Braconnot was crystallized from the acid hydrolyzate of muscle and wool, and named it leucine. L-leucine is one of the 18 common amino acids and one of the essential amino acids for the human body. Leucine can be used as a nutritional supplement and flavor enhancer. Amino acid infusions and comprehensive amino acid preparations, hypoglycemic agents, and plant growth promoters can be prepared. The production methods of L-leucine mainly include extraction method, chemical synthesis method, enzyme catalysis method, microbial fermentation method and so on. Most manufacturers in my country use protein hydrolysis to produce L-leucine, but the production of L-leucine by hydrolysis is time-consum...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/06C12R1/13
Inventor 刘帅李静宋朋睿
Owner HENAN JULONG BIOLOGICAL ENG CO LTD
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