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Fermentation process for increasing yield of L-arginine

A technology of arginine and technology, applied in the field of bioengineering, can solve the problems of unsatisfactory production level and output, low acid production level, high cost, etc.

Inactive Publication Date: 2014-01-29
SHANDONG MINQIANG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to produce L-arginine by microbial fermentation, the acid production level of most domestic fermentation production technologies is low and the cost is high, and the production level and output are far from meeting the needs of the domestic market.

Method used

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  • Fermentation process for increasing yield of L-arginine
  • Fermentation process for increasing yield of L-arginine
  • Fermentation process for increasing yield of L-arginine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] A kind of processing step of improving L-arginine fermentation acid production is as follows:

[0054] Strain selection:

[0055] From Brevibacterium flavum ATCC21493 (SD R ) as the starting bacteria, add a certain concentration of AHV (α-amino-β-hydroxyvaleric acid) or 2-TA (α-thiazolealanine) to the selective medium to induce TA174 (SD R 、AHV R 、Z-TA R ) strains;

[0056] TA188 (SD R 、AHV R , 2-TA R , L-His - ) strains;

[0057] After TA188 was treated with NTG, the mutant strain TA189 (SD R 、AHV R , 2-TA R , L-His - ) strains.

[0058] As preferably, the screening of the Brevibacterium flavum TA174:

[0059] Starting strain: Brevibacterium flavum ATCC214939 (SD R )

[0060] Described strain, the composition of the mutagenesis medium used for screening is:

[0061] culture medium

[0062] a. Complete medium: peptone 1.0%, beef extract 1.0%, yeast extract 0.5%, sodium chloride 0.5%, agar powder 2.0%, PH7.0-7.2 (test paper), sterilization temperature 1...

Embodiment 2

[0100] Screening of Brevibacterium flavum TA188 Mutant Strain and Fermentation in 30L Tank

[0101] (1), materials and methods

[0102] ①, starting strain Brevibacterium flavum AT174 (SD R 、AHV R , 2-TA R )

[0103] ②. Medium The selective medium is supplemented with 0.01% each of L-histidine (L-His), L-proline (L-Pro) and methionine (Met) on the basis of the basic medium.

[0104] ③. Mutagenesis method and selection of resistant bacteria

[0105] Centrifuge the value-added solution of the complete medium, wash the bacteria twice with 1 / 30M phosphate buffer and prepare a single-cell bacterial suspension, which is properly diluted and spread on a selective medium plate for 2-3 days at 30°C. The colonies that do not grow on the medium plate but grow on the selective plate may be L-His-deficient (L-His － ) or L-Pro deficiency (L-Pro － ) or Met-deficient (Met － ), and then confirmed by growth spectrum method.

[0106] Results and discussion

[0107] 1. The characteris...

example 1

[0116] Example 1. The initial volume (after planting) is 12L, the seed volume is 10%, the initial sugar is 12.5% ​​(1.5 kg of oral sugar), and the sugar supplement (1.8 kg of oral sugar) is made into about 3 L of 60% sugar solution, which contains C.S.L 60g acidic component After digestion, mix with sugar supplement.

[0117] Sugar supplement method: artificial intermittent or automatic pulse. Fermentation medium ammonium initial sulfide 4%, 18 h OD (1 / 100) is 0.47, when the wet bacteria body is 0.72g / 10ml, the pH is positioned at 6.6; take industrial glutenic acid (containing about 95%) and add water to adjust the pH to about 6.0 with ammonia water Dissolve and add 4 times after sterilization, 1% (120g) each time, 18 hours for the first time, and then add the second time when the residual L-GA is lower than 0.5% as measured by the Warburg respiration instrument.

[0118]

[0119] time (h) pH OD (1 / 100) Wet bacteria (g / 10ml) R.G (%) L-arg (%) 0 6.8 ...

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Abstract

The invention aims at providing a fermentation process for increasing the yield of L-arginine. The adopted technical scheme is as follows: brevibacterium flavum ATCC21493 (SD<R>) is adopted as a starting bacterium, a strain TA189 is obtained through NTG (Nitrosoguanidine) treatment and mutagenesis and can directly synthesize arginine by using glucose, the arginine production capacity is strong, the strain production capacity and the fermentation process are controlled to reach indexes, and the comprehensive level of the process reaches the leading domestic level. According to the fermentation process provided by the invention, the characteristics of the fermentation processes of amino acid and antibiotics are combined, the vitality of starting and ending thalli is strictly controlled by adopting trend control and a midway feed and glucose supplementing process, and a relatively high arginine production level is obtained.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a process for improving the fermentation of L-arginine to produce acid. Background technique [0002] L-arginine is one of the most important varieties of amino acids. It is widely used in the fields of medicine, nutrition and health care, and food, and has a large demand. It is useful for treating physiological functions, cardiovascular diseases, stimulating the immune system, maintaining nutritional balance of infants, It has good curative effect on promoting detoxification of the human body, and is called by experts as an important carrier for transporting and storing amino acids in the body, and is extremely important in intramuscular metabolism. L-arginine has the following functions: L-arginine is the main precursor of nitric oxide in the body, nitric oxide is an endogenous cell stretching factor, which can expand small arteries (such as penis blood vessels, making ...

Claims

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Application Information

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IPC IPC(8): C12P13/10C12N15/01C12N1/20C12R1/13
Inventor 高法民高树营李令娣王威
Owner SHANDONG MINQIANG BIOTECH
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