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Method for producing polymalic acid of different molecular weights through combined regulation

A technology of polymalic acid and molecular weight, applied in the field of microbial fermentation, can solve the problems of difficult extraction, low purity of β-polymalic acid, high cost of raw materials, etc., and achieve the effect of wide and reasonable application

Inactive Publication Date: 2014-01-15
天津北洋百川生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the use of organic nitrogen sources has problems such as high raw material costs, difficulty in later extraction, and low purity of the obtained β-polymalic acid.

Method used

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  • Method for producing polymalic acid of different molecular weights through combined regulation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A fermentation method for producing polymalic acid with different molecular weights, the steps are as follows:

[0025] 1. Composition of fermentation medium: sucrose 120g / L, NaNO 3 2g / L, KH 2 PO 4 0.1g / L, KCl0.4g / L, MnSO 4 0.05g / L, MgSO 4 ·7H 2 O1.0g / L, CaCO 3 20g / L, NH 4 NO 3 14.5g / L, aspartic acid 0.25~0.30g / L, leucine 0.58g / L, threonine 0.25g / L, valine 0.5g / L, histidine 0.35g / L, cytosine 0.001g / L, adenine 0.0015g / L, choline 0.1g / L, the rest is water, and the initial pH is 7.0.

[0026] 2. Activate the Aureobsidium pullulans CGMCC3336 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 cultivate at a constant temperature of 25°C and 200r / min 72h to prepare the seed solution; the composition of the seed medium is: 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, MgSO4 7H 2 O0.1g / L, KH 2 PO 4 1g / L, ZnSO 4 ...

Embodiment 2

[0030] A fermentation method for producing polymalic acid with different molecular weights, the steps are as follows:

[0031] 1. Composition of fermentation medium: sucrose 120g / L, NaNO 3 2g / L, KH 2 PO 4 0.1g / L, KCl0.4g / L, MnSO40.05g / L, MgSO 4 ·7H 2 O1.0g / L, CaCO 3 20g / L, NH 4 NO 3 14.5g / L, aspartic acid 0.30~0.35g / L, leucine 0.58g / L, threonine 0.25g / L, valine 0.5g / L, histidine 0.35g / L, cytosine 0.001g / L, adenine 0.0015g / L, choline 0.1g / L, the rest is water, and the initial pH is 7.0.

[0032] 2, the preparation of seed liquid and the bacterial classification used are basically the same as embodiment 1; L.K 2 CO 3 0.4g / L, MgSO 4 ·7H 2 O0.1g / L, KH 2 PO41g / L, ZnSO 4 ·7H 2 O0.05g / L, the rest is water, pH7.0.

[0033] 3. Connect the seed liquid obtained in step 2 to a 5L (3L liquid) fermenter at a volume ratio of 10%, with a ventilation rate of 1.0 (V / V), 400r / min, and culture at 25°C for 120h. Sampling in 2 hours, use amino column to detect the content of aspart...

Embodiment 3

[0036] A fermentation method for producing polymalic acid with different molecular weights, the steps are as follows:

[0037] 1. Composition of fermentation medium: sucrose 120g / L, NaNO 3 2g / L, KH 2 PO 4 0.1g / L, KCl0.4g / L, MnSO40.05g / L, MgSO 4 ·7H 2 O1.0g / L, CaCO 3 20g / L, NH 4 NO 3 14.5g / L, aspartic acid 0.35~0.4g / L, leucine 0.58g / L, threonine 0.25g / L, valine 0.5g / L, histidine 0.35g / L, cytosine 0.001g / L, adenine 0.0015g / L, choline 0.1g / L, the rest is water, and the initial pH is 7.0.

[0038] 2, the preparation of seed liquid and the bacterial classification used are basically the same as embodiment 1; L.K 2 CO 3 0.4g / L, MgSO 4 ·7H 2 O0.1g / L, KH 2 PO41g / L, ZnSO 4 ·7H 2 O0.05g / L, the rest is water, pH7.0.

[0039] 3. Connect the seed liquid obtained in step 2 to a 5L (3L liquid) fermenter at a volume ratio of 10%, with a ventilation rate of 1.0 (V / V), 400r / min, and culture at 25°C for 120h. Sampling at 2 hours, using amino column to detect aspartic acid conten...

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Abstract

The invention provides a method for producing polymalic acid of different molecular weights. The method comprises the following steps: inoculating an aureobasidium pullulans seed solution to a fermentation medium, and fermenting to produce polymalic acid; controlling a content of asparaginic acid in the fermentation medium to be 0.25-0.45g / L; regulating the pH value of the fermentation liquor to 6.0-7.0 in the fermentation process; and producing the polymalic acid of different molecular weights. The fermentation process is regulated, and the polymalic acid of different molecular weights is obtained, so that the polymalic acid is widely and reasonably applied to the fields of foods, medicines, chemical industry and petroleum according to different polymerization degrees of the polymalic acid. Moreover, according to a synergistic effect between the asparaginic acid and the pH value, the fermentation medium is further optimized, and the molecular weight of the produced polymalic acid can be accurately oriented in a narrow range, so that the requirements of people for the polymalic acid with different polymerization degrees are met.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a method for producing polymalic acid with different molecular weights by fermentation of Aureobasidium pullulans. Background technique [0002] Today, most synthetic polymers are petrochemical products, which have a profound impact on the environment. With the accumulation of people's environmental protection knowledge, the traditional production of non-degradable polymers has changed. At present, people are focusing on environmentally friendly biodegradable polymers. The production of these polymers is conducive to saving energy and resources, thereby slowing down the greenhouse effect and developing ecologically harmonious processes and products. [0003] Polymalic acid (polyhydroxysuccinate, Polymalic acid or Polymalate, PMLA for short) is a high molecular polymer synthesized with malic acid as the only monomer. Malic acid is a dicarboxylic acid containing ...

Claims

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Application Information

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IPC IPC(8): C12P7/62C12R1/645
Inventor 乔长晟宋玉民郑志达李政
Owner 天津北洋百川生物技术有限公司
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