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Method for producing L-glutamic acid by fermentation in staged gradient oxygen supply manner

A technology of glutamic acid and oxygen supply, which is applied in the field of microbial fermentation, can solve the problems of low oxygen utilization rate, achieve the effect of reducing power cost and operating cost, avoiding the accumulation of by-products, and high conversion rate of sugar and acid

Inactive Publication Date: 2013-07-24
北京轻发生物技术中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because oxygen is insoluble in water, the utilization rate of oxygen in the glutamic acid fermentation process is low, only 40%-60%

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Culture medium preparation

[0033] Incline medium (g / L): peptone 10, yeast powder 5, NaCl2.5, agar 30, pH7.0-7.2, 0.1MPa sterilization for 30min;

[0034] Seed medium: glucose 30g / L, corn steep liquor 40mL / L, soybean meal hydrolyzate 10mL / L, K 2 HPO 4 ·3H 2 O1.0g / L, MgSO 4 ·7H 2 O0.5g / L, urea 3g / L, pH7.0-7.2, 0.1Mpa sterilization for 15min;

[0035] Fermentation medium: glucose 80g / L, Na 2 HPO 4 12H 2 O2.5g / L, VB 1 0.4mg / L, MgSO 4 ·7H 2 O1g / L, MnSO 4 ·H 2 O2mg / L, FeSO 4 ·7H 2 O2mg / L, KCl1.3g / L, corn steep liquor 3ml / L, molasses 1.1g / L, medium pH value 7.0~7.2, 0.1Mpa sterilization for 15min;

[0036] 2. Fermentation

[0037] Corynebacterium glutamicum GKG-9 was inoculated in the above slant medium for activation, and cultured at 32°C for 20 hours to grow a lawn;

[0038] Pick the bacterial lawn from the freshly activated slant, inoculate it in the above-mentioned seed medium, shake and cultivate it at 37°C and 200r / min for 7-8h, and obtain the seed ...

Embodiment 2

[0042] Incline medium and seed medium formula are with embodiment 1;

[0043] Fermentation medium: glucose 75g / L, Na 2 HPO 4 12H 2 O2.5g / L, VB 1 1mg / L, MgSO 4 ·7H 2 O1.5g / L, MnSO 4 ·H 2 O1mg / L, FeSO 4 ·7H 2 O3mg / L, KCl2g / L, corn steep liquor 4ml / L, molasses 1.8g / L, medium pH value 7.0~7.2, 0.1Mpa sterilization for 15min;

[0044] In the fermentation process, except that the inoculum size is 5%, the dissolved oxygen concentration in the fermented liquid of the fermentation is controlled by a stepwise gradient oxygen supply mode, the initial ventilation rate is set to be 1L / min, the initial stirring speed is 500r / min, and by controlling Ventilation volume (1-2L / min) and stirring speed (500-880r / min) make the concentration of dissolved oxygen in the fermentation liquid of Corynebacterium glutamicum GKG-9 from growth inhibition phase to logarithmic growth phase 25-30% , the dissolved oxygen concentration of the glutamic acid-producing bacteria in its transition phase fer...

Embodiment 3

[0046] Incline medium and seed medium formula are with embodiment 1;

[0047] Fermentation medium: glucose 90g / L, Na 2 HPO 4 12H 2 O1g / L, VB 1 0.1mg / L, MgSO 4 ·7H 2 O0.5g / L, MnSO 4 ·H 2 O2.5mg / L, FeSO 4 ·7H 2 O1.5mg / L, KCl1g / L, corn steep liquor 2ml / L, molasses 0.5g / L, medium pH value 7.0-7.2, 0.1Mpa sterilization for 15min;

[0048] In the fermentation process, except that the inoculum size is 15%, the dissolved oxygen concentration in the fermented liquid of the fermentation is controlled by a stepwise gradient oxygen supply mode, the initial aeration rate is set to 2L / min, the initial stirring speed is 800r / min, and by controlling Ventilation volume (1-2L / min) and stirring speed (500-880r / min) make the concentration of dissolved oxygen in the fermentation liquid of Corynebacterium glutamicum GKG-9 from growth inhibition phase to logarithmic growth phase 40-45% , the dissolved oxygen concentration of the glutamic acid-producing bacteria in its transition phase ferm...

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Abstract

The invention discloses a method for producing an L-glutamic acid by fermentation in a staged gradient oxygen supply manner. The L-glutamic acid is obtained by inoculating a glutamic acid producing strain into a fermentation medium for fermentation and controlling the concentration of dissolved oxygen in a fermented liquid in a staged gradient oxygen supply manner. According to the method, the yield of the L-glutamic acid achieves over 155g / L and the glucose-acid conversion rate is raised to over 64% by means of respectively controlling different concentrations of dissolved oxygen at an inhibition stage, a logarithmic stage, a transformation stage and an acid production stage of the glutamic acid producing strain and through fed-batch fermentation; and the byproduct, namely lactic acid, during fermentation is reduced greatly.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for producing L-glutamic acid by fermentation. Background technique [0002] At present, the fermentation method is the main method to produce glutamic acid, and its production strains mainly include Corynebacterium glutamicum, Brevibacterium tianjinese, Corynebacterium crenalum, Corynebacterium Beijing pekinense), Brevibacterium flavum and their mutants, etc. However, the production level of glutamic acid fermentation depends not only on the performance of the strain itself, but also must be given suitable environmental conditions to fully exert and show their excellent production capacity, such as medium composition, temperature, solubility, and so on. Oxygen, feed, etc. [0003] The Chinese invention patent with the application number 91107357.4 discloses a new L-glutamic acid fermentation process. In order to improve the acid production rate and conversion rat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/14C12Q3/00C12R1/13C12R1/15
Inventor 邱博谢希贤张成林万红兵欧阳宇红曹文杰
Owner 北京轻发生物技术中心
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