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Method for producing L-leucine by fermentation

A leucine and fermenter technology, applied in the field of fermentation and production of L-leucine, can solve problems such as high cost, unsuitable for large-scale production, and cumbersome procedures

Active Publication Date: 2019-12-06
XINJIANG FUFENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The applicant's previous patented technology "a fermentation process for producing L-leucine" increases the production of leucine by adding compound amino acid solution and nutrient solution in the middle and late stages of fermentation, but the cost of the above-mentioned feeding solution is relatively high , not suitable for large-scale production
The applicant's previous patented technology "a method for increasing the fermentation yield of L-leucine" adopts membrane-coupled batch dialysis fermentation, which effectively reduces the concentration of by-product and improves the conversion rate of sugar and acid , by supplementing the nutrient solution containing glycerol and betaine in the middle stage of fermentation, the production of L-leucine and the conversion rate of sugar and acid in the fermentation broth can be promoted; however, this method has the disadvantage of complicated procedures

Method used

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  • Method for producing L-leucine by fermentation
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  • Method for producing L-leucine by fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A method for producing L-leucine by fermentation, comprising the steps of:

[0029] Test strain: Brevibacterium flavum ( Brevibacterium flavum ) TK0303 (Met - +Ile L +2-TA r +α-AB r +β-HL r +Rif r +SG r ).

[0030] The Brevibacterium flavum seed solution was obtained by conventional culture methods, and the Brevibacterium flavum seed solution was inserted into a 500L fermenter equipped with a 300L fermenter medium at an inoculation amount of 10%, and the inoculation concentration was OD 600 1.5; the stirring speed is 400r / min; the dissolved oxygen is controlled at 25% by stirring and ventilation; the pH is controlled at 7.0 by automatic flow of ammonia water; the cultivation temperature is 33°C; Feed into the tank, maintain the sugar concentration in the fermentation tank at 1g / L, stop feeding 6 hours before the end of fermentation; the fermentation period is 44h.

[0031] Components of the fermenter culture medium: glucose 50g / L, (NH 4 ) 2 SO 4 10g / L, cor...

Embodiment 2

[0037] A method for producing L-leucine by fermentation, comprising the steps of:

[0038] Test strain: Brevibacterium flavum ( Brevibacterium flavum ) TK0303 (Met - +Ile L +2-TA r +α-AB r +β-HL r +Rif r +SG r ).

[0039] The Brevibacterium flavum seed solution was obtained by conventional culture methods, and the Brevibacterium flavum seed solution was inserted into a 500L fermenter equipped with a 300L fermenter medium at an inoculation amount of 8%, and the inoculation concentration was OD 600 1.8; the stirring speed is 350r / min; the dissolved oxygen is controlled at 25% by stirring and ventilation; the pH is controlled at 6.8 by automatic flow of ammonia water; the cultivation temperature is 33°C; Feed into the tank, maintain the sugar concentration in the fermentation tank at 1g / L, stop feeding 6 hours before the end of fermentation; the fermentation period is 44h.

[0040] Components of the fermenter culture medium: glucose 50g / L, (NH 4 )2 SO 4 10g / L, corn s...

Embodiment 3

[0050] 1. On the basis of the conventional fermentation of Comparative Example 1, the effect of the added concentration of sodium acetate on the fermented liquid was checked. L- Leu and the influence of the main by-product content. Set the concentration gradient of sodium acetate in the fermentation broth, respectively 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, the unit is g / L. like Figure 1-2 As shown, with the increase of sodium acetate concentration, the L- The content of Leu increases steadily, while the content of by-products Val and Hac gradually decreases. When the concentration of sodium acetate increases to 0.5g / L, the content of L-Leu reaches the maximum value. Continue to increase the content of sodium acetate, and the content of L-Leu does not increase At this time, the content of the by-products Val and Hac did not continue to decrease, indicating that at this time, the metabolic fluxes have reached equilibrium. During the fermentation process, by adding an appropr...

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Abstract

The invention belongs to the field of production process of L-leucine, and discloses a method for producing L-leucine by fermentation. The method comprises the following steps: fermenting brevibacterium flavum producing L-leucine for more than 44 h and feeding a sodium acetate aqueous solution into a fermentation tank at 30 h of the fermentation, controlling concentration of sodium acetate at 0.1-1 g / L in the fermentation solution until the fermentation is completed, wherein the concentration of the sodium acetate aqueous solution is 50-100 g / L; feeding a malonic acid aqueous solution in the fermentation tank at 30 h of the fermentation and controlling concentration of malonic acid at 0.1-0.5 ml / L in the fermentation solution until the fermentation is completed, wherein the concentration of the malonic acid aqueous solution is 10-20%; adding chitosan in the fermentation tank at 40 h of the fermentation and controlling concentration of chitosan at 20-80 mg / L in the fermentation solution. The method of the invention has high fermentation efficiency, simple operation and low cost.

Description

technical field [0001] The invention belongs to the field of L-leucine production technology, and specifically provides a method for fermenting and producing L-leucine. Background technique [0002] L-leucine is also called leucine, its chemical name is α-aminoisocaproic acid, and its molecular formula is C 6 h 13 o 2 N, with a relative molecular weight of 131.18, is a non-polar amino acid with a slightly bitter taste, soluble in water, and an isoelectric point of 5.98. It is widely used in many industries such as medicine, food, seasoning, animal feed and cosmetics. my country's research on L-leucine started relatively late, some enterprises produced low levels of L-leucine by fermentation, and there were relatively few studies on downstream pretreatment and extraction processes, it was difficult to realize industrial production, and there were Problems such as high acid and alkali consumption, serious pollution, and low yield and purity of the current product. [0003] ...

Claims

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Application Information

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IPC IPC(8): C12P13/06C12R1/13
CPCC12P13/06
Inventor 冯世红包鑫史静东韦树高边恩来
Owner XINJIANG FUFENG BIOTECH
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