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Method for producing L-diaminocaproic acid through multistage continuous fermentation

A lysine and fermenter technology, applied in the field of L-lysine preparation, can solve the problems of reducing the utilization rate of the product, affecting the yield of the product, etc., and achieve the advantages of high utilization rate of the substrate, stable yield and lower production cost. Effect

Inactive Publication Date: 2016-11-23
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the strains used in the production of L-lysine at home and abroad are mainly modified strains of Corynebacterium glutamicum, Brevibacterium lactofermentum, Escherichia coli and Brevibacterium flavum. Inhibition, which reduces the utilization rate of the product and affects the yield of the product

Method used

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  • Method for producing L-diaminocaproic acid through multistage continuous fermentation
  • Method for producing L-diaminocaproic acid through multistage continuous fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0028] A method for producing L-lysine by multistage continuous fermentation, comprising the following steps:

[0029] Step 1, prepare shake flask medium, seed medium, initial fermentation medium, carbon source feed tank culture fluid, nitrogen source feed tank culture fluid and medium feed tank culture fluid, the formula is as follows:

[0030] Shake flask medium: sucrose 8g / L (dissolved), ammonium sulfate 2g / L, yeast extract 5g / L, peptone 10g / L, K 2 HPO 4 .3H 2 02g / L, monosodium glutamate 2g / L, pyruvic acid 0.5g / L, MgSO 4 0.4g / L, FeSO 4 .7H 2 0 0.01g / L, L-threonine 0.2g / L, biotin 5mg / L. PH6.8-7.0.

[0031] Seed medium: sucrose 25g / L (divided), yeast extract 5g / L, ammonium sulfate 10g / L, K 2 HPO 4 .3H 2 0 1g / L, MSG 2.5g / L, pyruvic acid 0.5g / L, MgSO 4 0.4g / L, FeSO 4 .7H 2 0 0.01g / L, L-threonine 0.2g / L, biotin 2.5mg / L, nicotinamide 1.5mg / L, thiamine 0.45mg / L, pH6.8-7.0.

[0032] Initial fermentation medium: glucose 40 g / L (divided), ammonium sulfate 20 g / L, potas...

Embodiment 2

[0040] Steps 1-3 are the same as in Example 1, wherein the sugar used in the fermentation medium and the carbon source medium is starch saccharification solution.

[0041] Step 4: After connecting the sterilized fermentation tanks in series, insert 600ml of initial fermentation medium into the first-level fermentation tank, and feed in sterile air with an air volume of 0.04 / (min L), according to the inoculum size of 10% (v / v) Connect the first-level seed tank to cultivate mature strains, and the initial fermentation medium flows from the first-level fermentation tank into the second, third, and fourth levels. Use carbon source feeding tanks and medium feeding tanks to control the inflow of fermentation broth The rate is 100-120ml / h, the flow rate of the carbon source feed medium is controlled to 30ml / h, and the outflow rate of the fermentation broth is 130-150ml / h, so that the initial sugar concentration in the primary fermentation tank is 80g / L. During the continuous fermentat...

Embodiment 3

[0043] Steps 1-3 are the same as in Example 2.

[0044] Step 4: After connecting the sterilized fermentation tanks in series, insert 600ml of initial fermentation medium into the first-level fermentation tank, and inject sterile air with an air volume of 0.02L / (min·L), according to the inoculum size of 10 %(v / v) is connected to the first-level seed tank to cultivate mature strains, and the initial fermentation medium flows from the first-level fermenter to the second, third and fourth levels, and the carbon source feeding tank and the medium feeding tank are used to control the fermentation liquid The inflow rate is 50-70ml / h, the flow acceleration rate of the carbon source feeding tank culture fluid is 20-30ml / h, and the outflow rate of the fermentation broth is 80-100ml / h, so that the initial sugar concentration in the primary fermentation tank is 50g / L , in the continuous fermentation process, the sugar concentration in the fermentation tanks at all levels is as follows: fi...

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Abstract

The invention discloses a method for producing L- diaminocaproic acid through multistage continuous fermentation. The method includes: starting from escherichia coil to obtain an L-diaminocaproic product through fermentation above stage four; specifically stringing at least four fermentation tanks integrally, and controlling carbon source concentration and nitrogen source concentration in the fermentation tanks of different stages by regulating ventilatory capacity, initial sugar concentration of the primary fermentation tank, and addition rates, fermented-liquid outflowing rate and fermented liquid inflowing rate of carbon source filling containers and nitrogen source filling containers of the fermentation tanks; controlling the fermentation tanks of different stages to enable the specific growth rate of thallus of the fermentation tanks of the different stages to be 0.3-0.8h<1> to obtain the fermentation product. With the method, high thalli concentration in the fermentation system is maintained, lag phase of cells is shortened, substrate utilization is higher as compared with batch fermentation, target product quantity is high, yield is stable, operations of repeated tank washing, sterilizing, inoculating and the like are not needed, and production cost is lowered.

Description

technical field [0001] The invention belongs to the field of L-lysine preparation, and in particular relates to a method for producing L-lysine by multistage continuous fermentation. Background technique [0002] The chemical name of lysine is 2,6-diaminocaproic acid (2,6- diaminohexanoic acid ), molecular formula NH 2 (CH 2 ) 4 CH. (NH 2 )COOH, with a molecular weight of 146.19, is one of the nine essential amino acids for living organisms. There are two types of lysine in nature: L-type and D-type, and L-lysine can be used by humans and animals. Lysine can regulate the balance of metabolism in the body, improve the body's absorption of cereal protein, improve human dietary nutrition and animal nutrition, and promote growth and development; it can also promote the secretion of digestive enzymes, promote the delivery of intracellular calcium, and improve the metabolism of nitrogen in the body balance. [0003] Lysine is the second most productive commercial amino acid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/08C12R1/19
CPCC12N1/20C12P13/08
Inventor 陈可泉齐雁斌张阿磊陈西欧阳平凯
Owner NANJING UNIV OF TECH
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