Method for producing polymalic acid by high-density fermentation

A polymalic acid and fermenter technology, which is applied in the field of microbial fermentation to achieve the effects of reducing equipment scale, saving power consumption and energy consumption, and improving production intensity

Active Publication Date: 2016-03-02
天津北洋百川生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Regarding the fermentation of polymalic acid, most of the literatures increase the yield of polymalic acid through strain mutagenesis and medium optimization, and rarely mention that the production of polymalic acid can be regulated and improved by controlling the fermentation conditions. The invention patent discloses a new process for the production of β-polymalic acid by microbial fermentation. It is mentioned in the article that ventilation, tank pressure, stirring rate and dissolved oxygen concentration should be controlled during the fermentation process. Therefore, a new method is sought. To further increase the production of polymalic acid is a major problem to be solved at present

Method used

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  • Method for producing polymalic acid by high-density fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] (1) Activate the Aureobsidium pullulans CGMCC.NO.3336 strain for 6-8 hours, that is, transfer it from the sucrose slant medium to a 500ml baffle bottle with a liquid volume of 50ml and a constant temperature of 25°C. Cultivate at 200r / min for 72h to prepare seed liquid;

[0025] The seed culture medium is composed of: sucrose 140g / L, yeast extract 3g / L, ammonium sulfate 1g / L, succinic acid 2g / L, corn steep liquor 1g / L, K 2 CO 3 0.4g / L, MgSO 4 ·7H 2 O0.1g / L, KH 2 PO 4 1g / L, ZnSO 4 ·7H 2 O0.05g / L, the rest is water, pH7.0.

[0026] (2) Connect the obtained seed liquid to a 5L fermentor at 10% (v / v) for fermentation and culture, according to the OD of the bacteria 600 Value controls pH and dissolved oxygen:

[0027] The first stage of fermentation: when the bacteria OD 600 When it is 0.67, the dissolved oxygen is reduced to 55%, and the stirring is connected in series to maintain the dissolved oxygen at 50%. At the same time, the pH is adjusted to 7.0 with hydrochloric acid and s...

Embodiment 2

[0032] The activation of the strain and the preparation of the seed liquid are the same as in Example 1

[0033] (2) Connect the obtained seed liquid to a 5L fermentor at 10% (v / v) for fermentation and culture, according to the OD of the bacteria 600 Value controls pH and dissolved oxygen:

[0034] The first stage of fermentation: when the bacteria OD 600 When it is 0.65, the dissolved oxygen is reduced to 60%, and the stirring is connected in series to maintain the dissolved oxygen at 55%. At the same time, the pH is adjusted to 6.8 with hydrochloric acid and sodium hydroxide to promote the growth of bacteria;

[0035] The second stage of fermentation: when the bacteria OD 600 When it is 0.75, the dissolved oxygen is reduced to 32%, the temperature is 25°C, and the pH is adjusted to 5.0 with hydrochloric acid and sodium hydroxide, and the fermentation is 104 hours to promote the large-scale synthesis of polymalic acid by the bacteria;

[0036] The concentration of hydrochloric acid is...

Embodiment 3

[0039] The activation of the strain and the preparation of the seed liquid are the same as in Example 1

[0040] (2) Connect the obtained seed liquid to a 5L fermentor at 10% (v / v) for fermentation and culture, according to the OD of the bacteria 600 Value controls pH and dissolved oxygen:

[0041] The first stage of fermentation: when the bacteria OD 600 When it is 0.63, the dissolved oxygen is reduced to 62%, and the stirring is connected in series to maintain the dissolved oxygen at 58%. At the same time, the pH is adjusted to 6.0 with hydrochloric acid and sodium hydroxide to promote the mass growth of bacteria;

[0042] The second stage of fermentation: when the bacteria OD 600 When it is 0.72, the dissolved oxygen is reduced to 35%, the temperature is 25°C, and the pH is adjusted to 4.9 with hydrochloric acid and sodium hydroxide at the same time, and the fermentation is 112 hours to promote the large-scale synthesis of polymalic acid by the bacteria;

[0043] The concentration o...

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Abstract

The invention discloses a method for high-density fermentation production of polymalic acid and belongs to the field of microbial fermentation. According to the method for high-density fermentation production of the polymalic acid, pH and dissolved oxygen are regulated and controlled in two stages, and in the first stage, mass growth of thalli is realized; and in the second stage, the pH and the dissolved oxygen are changed to realize mass accumulation of the polymalic acid. The invention provides the method for production of the polymalic acid, the operation is simple, the effects are obvious, the fermentation period is shortened, and the production efficiency of the polymalic acid is further greatly improved; and according to the two-stage process for high-density fermentation, on the one hand, the scale of equipment in the thallus culture stage can be reduced, and the power consumption and the energy consumption of a large-scale fermentation tank in the fermentation process are saved, and on the other hand, the production intensity for cell culture and synthesis of the polymalic acid can be improved, and the production process can achieve the purposes of saving energy and reducing consumption.

Description

Technical field [0001] The invention belongs to the field of microbial fermentation, and specifically relates to a method for producing polymalic acid by high-density fermentation, and particularly relates to a method for producing polymalic acid by regulating pH and dissolved oxygen in two stages. Background technique [0002] Now, most synthetic polymers are petrochemical products, which have a profound impact on the environment. With the accumulation of people's knowledge of environmental protection, the traditional non-degradable polymer production has changed. At present, the focus of attention is on environmentally friendly biodegradable polymers. The production of these polymers is conducive to saving energy and resources, thereby slowing the greenhouse effect, and developing ecologically coordinated processes and products. [0003] Polymalic acid (Polymalicacid or Polymalate, referred to as PMLA) is a homopolymeric polymer synthesized with malic acid as the only monomer. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/62C12R1/645
Inventor 乔长晟宋玉民范栩嘉郝华旋
Owner 天津北洋百川生物技术有限公司
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