A high-yield polymalic acid Aureobasidium pullulans and its application

A technique of Aureobasidium pullulans and polymalic acid, applied in the direction of microorganism-based methods, fungi, microorganisms, etc., can solve the problems of high raw material cost, difficult quality control, low yield of polymalic acid, etc., and meet the simple nutritional requirements of strains , reduce the consumption of cooling water, the effect of significant economic benefits

Active Publication Date: 2020-06-02
GUANGXI UNIV
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Problems solved by technology

[0006] (1) High-concentration glucose is used as a carbon source. On the one hand, the cost of raw materials is higher, and on the other hand, high-concentration glucose has a substrate inhibitory effect on the growth of the thallus, thereby limiting the improvement of polymalic acid output; and when molasses is used as a carbon source, due to Molasses itself has a darker color and more impurities. Although the cost of raw materials has decreased, it will increase the cost of extraction and purification of polymalic acid and treatment of fermentation waste liquid
[0007] (2) The yield of polymalic acid is still low. Although the maximum yield of polymalic acid synthesized by Aureobasidium pullulans ZD-3D can reach 62.27g / L, sodium fumarate needs to be added as a fermentation accelerator in the medium, which increases the fermentation rate. Raw material costs
[0008] (3) A certain amount of corn steep liquor or corn extract must be added to the above-mentioned three kinds of fermentation medium of Aureobasidium pullulans, which increases the cost of fermentation raw materials on the one hand, and is difficult to control due to the complexity of corn steep liquor or corn extract on the other hand. , there is uncertainty about the effect on the fermentation process
[0009] (4) The fermentation temperature of the three above-mentioned Aureobasidium pullulans is 25°C, and microbial fermentation is an exothermic process, so a large amount of cooling water is required to maintain a lower fermentation temperature, thereby increasing the operating cost of fermentation equipment

Method used

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  • A high-yield polymalic acid Aureobasidium pullulans and its application
  • A high-yield polymalic acid Aureobasidium pullulans and its application
  • A high-yield polymalic acid Aureobasidium pullulans and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 : Screening and Identification of Strains

[0034] 1.1 Screening process of strain GXZ-6

[0035] Enrichment culture: Weigh 5g of fresh leaf samples into a 250mL Erlenmeyer flask filled with 50mL of enrichment medium, culture at 30°C for 2 days at a shaker speed of 180rpm, to obtain a bacterial suspension. The concentration components of the enrichment medium are as follows: mannitol 100g / L, ammonium nitrate 1g / L, citric acid 2g / L, KH 2 PO 4 0.5g / L, MgSO 4 ·7H 2 O 0.2g / L, Tween-80 0.2g / L, distilled water to 1L, natural pH.

[0036] Bacterial suspension dilution and colony picking: The bacterial suspension obtained above was diluted into bacterial suspensions of different concentrations using the 10-fold dilution method, and 10 4 , 10 5 , 10 6 100 μL of the bacterial suspension was spread on the PDA plate medium, incubated at 30°C for 2 days, and yeast-like viscous colonies were picked. The concentration and components of the PDA medium are as follow...

Embodiment 2

[0051] Example 2 : Method for producing polymalic acid by fermentation of Aureobasidium pullulans GXZ-6 bacterial strain

[0052] (1) Strain activation: Aureobasidium pullulans GXZ-6 preserved in glycerin was streaked on a PDA slant medium, and cultured at a constant temperature of 28° C. for 3 days;

[0053] (2) Seed culture: Take one ring of activated slant strains, inoculate in a 500mL Erlenmeyer flask with 50mL liquid seed medium, shaker speed 160rpm, and cultivate at 25°C for 48h to obtain seed liquid; the concentration of liquid seed medium The components are as follows: glucose 80g / L, sodium nitrate 2g / L, KCl 0.4g / L, KH 2 PO 4 0.05g / L, MgSO 4 ·7H 2 O 0.1g / L, ZnSO 4 ·7H 2 O 0.05g / L, CaCO 3 15g / L;

[0054] (3) Fermentation culture: the seed liquid is inserted in the 250mL Erlenmeyer flask that 30mL liquid fermentation medium is housed according to 8% inoculation amount, shaker rotating speed 160rpm, 25 ℃ cultivates 9 days; The concentration component of liquid f...

Embodiment 3

[0058] Example 3 : Method for producing polymalic acid by fermentation of Aureobasidium pullulans GXZ-6 bacterial strain

[0059] Aureobasidium pullulans (Aureobasidium pullulans) GXZ-6 fermentation method for producing polymalic acid, the specific steps are:

[0060] (1) Strain activation: Aureobasidium pullulans GXZ-6 preserved in glycerol was streaked on the PDA slant medium, and cultured at 30° C. for 2 days;

[0061] (2) Seed culture: Take one ring of activated slant strains, inoculate in a 500mL Erlenmeyer flask with 100mL liquid seed medium, shaker speed 220rpm, cultivate at 30°C for 36h to obtain seed liquid; the concentration of liquid seed medium The components are as follows: glucose 100g / L, sodium nitrate 4g / L, KCl 0.6g / L, KH 2 PO 4 0.2g / L, MgSO 4 ·7H 2 O 0.3g / L, ZnSO 4 ·7H 2 O 0.2g / L, CaCO 3 30g / L;

[0062] (3) Fermentation culture: the seed liquid is inserted in the 250mL Erlenmeyer flask that 40mL liquid fermentation medium is housed according to 10%...

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Abstract

The invention belongs to the technical field of microorganism application, and in particular relates to Aureobasidium pullulans and its application in producing polymalic acid. A high-yield polymalic acid Aureobasidium pullulans, which is classified as Aureobasidium pullulans GXZ‑6, and the preservation number is CCTCC NO: M 2017517, and the preservation date is September 20, 2017. The preservation unit: China Center for Type Culture Collection, deposit address: Wuhan University, Wuhan, China. Aureobasidium pullulans (Aureobasidium pullulans) GXZ‑6 provided by the present invention can utilize tapioca starch, corn starch, wheat starch, soluble starch, starch saccharification liquid, maltose syrup as carbon source fermentation to produce polymalic acid, compared with utilizing glucose as For the production of polymalic acid by fermentation of carbon sources, the cost of raw materials is lower, and there is no problem that high-concentration carbon sources can inhibit the growth of bacteria.

Description

technical field [0001] The invention belongs to the technical field of microorganism application, and in particular relates to Aureobasidium pullulans and its application in producing polymalic acid. Background technique [0002] Polymalic acid [poly(β-L-malic acid), PMLA] is a new type of water-soluble polymer compound, which is formed by polymerizing malic acid as the only monomer through the linkage of α-hydroxyl and β-carboxyl through ester bonds. It has good chemical plasticity, biodegradability and biocompatibility, and has broad application prospects in food, medicine, agriculture and other fields. In 1969, polymalic acid was discovered for the first time in Penicillium cyclopium. Foreign researchers subsequently conducted extensive research on the preparation method, properties and uses of polymalic acid. It was not until around 2000 that there were related studies in China. research report. [0003] At present, polymalic acid is mainly prepared by microbial fermen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P7/62C12R1/645
CPCC12P7/625C12N1/145C12R2001/645
Inventor 曾伟梁智群陈桂光张斌李梦璇
Owner GUANGXI UNIV
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