Method for increasing the production of Paenibacillus polymyxa 2,3-butanediol by using vitamin C

A technology of spore-like polymyxa and vitamins, which is applied in the field of fermentation engineering, can solve problems such as impact, and achieve the effect of reducing accumulation obstruction and promoting glucose consumption

Inactive Publication Date: 2015-10-28
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to exogenous reducing agents (such as NaBH 4 and DTT) can affect the oxidation-reduction state of the cell environment. At present, the addition of exogenous reducing agents can regulate the oxidation-reduction potential of the external environment of the organism, thereby affecting the intracellular coenzyme NADH / NAD + There have been reports on promoting the accumulation of target products such as 1,3-propanediol, but the use of exogenous reducing agent VC to increase the production of (R,R)-2,3-butanediol has not yet been seen.

Method used

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  • Method for increasing the production of Paenibacillus polymyxa 2,3-butanediol by using vitamin C
  • Method for increasing the production of Paenibacillus polymyxa 2,3-butanediol by using vitamin C
  • Method for increasing the production of Paenibacillus polymyxa 2,3-butanediol by using vitamin C

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] A method utilizing vitamin C to improve Paenibacillus polymyxa 2,3-butanediol production, comprising the steps of:

[0019] (1) Preparation of medium

[0020] Solid slant medium composition in g / L: starch 20, glucose 5, peptone 2, MgSO 4 .7H 2 O0.5, NaCl0.5, corn steep liquor 2, agar 20, steam sterilization at 121°C for 20min;

[0021] Seed medium composition in g / L: starch 30, glucose 5, yeast extract powder 2, peptone 4, MgSO 4 .7H 2 O0.5, NaCl0.5, K 2 HPO 4 1.5, steam sterilization at 121°C for 20 minutes;

[0022] Fermentation medium components in g / L: glucose 55, yeast extract powder 10, NaCl5, K 2 HPO 4 4. KH 2 PO 4 2, (NH4) 2 SO 4 2, sodium citrate 2, MgSO 4 ·7H 2 O0.7, KCl0.5, CaCl 2 0.05, ZnSO 4 ·7H 2 O0.005, FeSO 4 ·7H 2 O0.005, MnSO 4 ·H 2 O0.005, pH 7.0, steam sterilization at 121°C for 20 minutes;

[0023] (2) Shake flask culture

[0024] Primary Seed Cultivation

[0025] Insert CJX-518 from the solid slant medium into 50mL seed medi...

Embodiment 2

[0032] Control group: Fermentation without vitamin C addition

[0033] (1)~(2) are the same as Steps (1)~(2) of Example 1;

[0034](3) Put the secondary seed culture solution into a 5L fermenter with 3L fermentation medium at an inoculation density of 10% by volume, and ferment at a ventilation rate of 0.2vvm, a stirring speed of 300rpm, and a temperature of 37°C. 24h, take the fermentation broth at 12000rpm and centrifuge for 10min at 12h, 18h, and 24h respectively, then take the supernatant, dilute it with ultrapure water 10 times, and filter it with a 0.22μm filter. The total amount of glucose as figure 1 As shown in the control group, the concentration of acetoin produced was as figure 2 As shown in the control group, the concentration of (R,R)-2,3-butanediol produced was as image 3 shown in the control group.

Embodiment 3

[0036] A method utilizing vitamin C to improve Paenibacillus polymyxa 2,3-butanediol production, comprising the steps of:

[0037] (1) Seed culture: cultivate CJX-518 in seed medium to obtain first-grade seeds; cultivate first-grade seeds in seed medium to obtain second-grade seeds; the preparation of seed medium is the same as in Example 1;

[0038] (2) Fermentation culture

[0039] Insert the secondary seeds into the fermentation medium at an inoculation density of 5% by volume. When the ventilation rate is 0.8vvm, the stirring speed is 500rpm, the temperature is 42°C, and the fermentation time is 12h, when the glucose in the fermentation medium is detected When the concentration is reduced to 2g / L, add a vitamin C aqueous solution with a concentration of 80g / L, the ratio of the amount of vitamin C added each time to the volume of the fermentation medium is 160mg:1L, and vitamin C is added every 1h until the end of fermentation. The content of (R,R)-2,3-butanediol in the fe...

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Abstract

The invention discloses a method for increasing the yield of paenibacillus polymyxa 2,3-butanediol by using vitamin C. The method comprises the following steps: (1) seed culture; (2) fermentation culture: inoculating secondary seeds into a fermentation culture medium, performing fermentation under the conditions that the ventilatory capacity is 0.1-0.8vvm, the stirring rotating speed is 300-500rpm and the temperature is 32-42 DEG C for 12-24 hours, and adding 80-160g / L vitamin C solution when the concentration of glucose in the fermentation culture medium is reduced to 2-10g / L. By the adoption of a method for continuously adding a reducing agent, the method disclosed by the invention is favorable for maintenance of a redox situation of an external environment in which bionts exist in the whole fermentation process, so that consumption of the glucose in fermentation liquid and conversion of acetoin are finally promoted, and the accumulation of (R,R)-2,2- butanediol is improved.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering and relates to a method for improving the 2,3-butanediol output of Paenibacillus polymyxa by using vitamin C. Background technique [0002] As an important chemical raw material and liquid fuel, 2,3-butanediol is widely used in the fields of medical treatment, chemical industry, energy and food, and has broad industrial application prospects. Paenibacillus polymyxa produces (R,R)-2,3-butanediol by biocatalyzing acetoin, because each molecule of acetoin is converted into (R,R)-2,3-butanediol A molecule of NADH needs to be consumed in the process of alcohol, therefore, ensuring sufficient supply of coenzyme NADH is conducive to the accumulation of (R,R)-2,3-butanediol. At present, in the process of promoting the conversion of acetoin into (R,R)-2,3-butanediol, the construction of a cofactor bioregeneration system has become the mainstream of research. Due to the relatively cumbers...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/18C12R1/01
Inventor 程景胜代军军元英进完莉萍赵琰
Owner TIANJIN UNIV
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