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Method for increasing yield 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: 2014-04-02
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 yield of paenibacillus polymyxa 2,3-butanediol by using vitamin C
  • Method for increasing yield of paenibacillus polymyxa 2,3-butanediol by using vitamin C
  • Method for increasing yield of paenibacillus polymyxa 2,3-butanediol by using vitamin C

Examples

Experimental program
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Effect test

Embodiment 1

[0018] A method for increasing the production of Bacillus polymyxa 2,3-butanediol by using vitamin C, comprising the following steps:

[0019] (1) Preparation of culture medium

[0020] The composition of the solid slant medium is calculated 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 20 min;

[0021] The composition of the seed medium is calculated in g / L: starch 30, glucose 5, yeast extract 2, peptone 4, MgSO 4 .7H 2 O0.5, NaCl0.5, K 2 HPO 4 1.5, 121℃ steam sterilization for 20min;

[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 20min;

[0023] (2) Shake flask culture

[0024] Cultivation of primary seeds

[0025] Connect CJX-518 from the solid slant culture...

Embodiment 2

[0032] Control group: fermentation without vitamin C

[0033] (1)~(2) Same as steps (1)~(2) of Example 1;

[0034] (3) Connect the secondary seed culture solution to a 5L fermentor equipped with 3L fermentation medium according to the volume percentage of 10% inoculation density, and ferment under the condition of aeration of 0.2vvm, stirring speed of 300rpm and temperature of 37℃ After 24h, take the fermentation broth at 12h, 18h, 24h, centrifuge at 12000rpm for 10min, take the supernatant, dilute with ultrapure water 10 times, filter with 0.22μm filter, high performance liquid chromatography to detect the content of fermentation product, the consumption detected in each time period The total amount of glucose such as figure 1 As shown in the control group, the concentration of acetoin produced is as figure 2 As shown in the control group, the concentration of (R,R)-2,3-butanediol produced is as follows image 3 Shown in the control group.

Embodiment 3

[0036] A method for increasing the production of Bacillus polymyxa 2,3-butanediol by using vitamin C, comprising the following steps:

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

[0038] (2) Fermentation culture

[0039] Connect the secondary seeds to the fermentation medium at an inoculum density of 5% by volume. When the aeration rate is 0.8vvm, the stirring speed is 500rpm, the temperature is 42℃, and the fermentation time is 12h, when the glucose in the fermentation medium is detected When the concentration is reduced to 2g / L, add an aqueous solution of vitamin C with a concentration of 80g / L. The ratio of the amount of vitamin C to the volume of the fermentation medium is 160mg:1L each time, and vitamin C is added once every 1h until the end of the fermentation. The ...

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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 production of Bacillus polymyxa 2,3-butanediol by using vitamin C. Background technique [0002] 2,3-Butanediol, as an important chemical raw material and liquid fuel, is widely used in medical, chemical, energy and food fields, and has broad industrial application prospects. Bacillus polymyxa produces (R,R)-2,3-butanediol through biocatalysis of acetoin, because each molecule of acetoin is converted into (R,R)-2,3-butanediol One molecule of NADH needs to be consumed in the alcohol process. Therefore, ensuring sufficient supply of coenzyme NADH is beneficial to the accumulation of (R,R)-2,3-butanediol. At present, in the process of promoting the conversion of acetoin to (R,R)-2,3-butanediol, the construction of a biological regeneration system of cofactors has become the mainstream of research. Due to the relatively cumbersome operating metho...

Claims

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Application Information

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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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