Aureobasidium pullulans and method for fermenting to produce pulullan thereof

A technology of Aureobasidium pullulans and pullulan polysaccharide is applied in the field of microorganisms to achieve the effects of reducing production costs and achieving obvious effects

Active Publication Date: 2018-02-16
INST OF AGRO FOOD SCI & TECH SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the use of ARTP to mutate the pullulan polysacchar

Method used

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  • Aureobasidium pullulans and method for fermenting to produce pulullan thereof
  • Aureobasidium pullulans and method for fermenting to produce pulullan thereof
  • Aureobasidium pullulans and method for fermenting to produce pulullan thereof

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Experimental program
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Embodiment 1

[0025] The identification of embodiment 1 original bacterial strain

[0026] The original strain Aureobasidium pullulans was purchased from China General Microorganism Culture Collection and Management Center, and the preservation number of the strain is AS3.3984.

[0027] Amplification and sequencing of ITS characteristic sequence and 18S rRNA gene sequence showed that the sequence similarity between strain AS3.3984 and Aureobasidium pullulans strain YY16 was 99%, and it was determined to be Aureobasidium pullulans. Sequencing was completed by Jinweizhi Biotechnology (Beijing) Co., Ltd.

Embodiment 2

[0029] ARTP mutagenesis of Aureobasidium pullulans AS3.3984 and screening of high-yield mutants

[0030] 2.1 Activation of the original strain

[0031] Prepare slant medium: glucose 15-20g / L, yeast extract powder 8-10g / L, peptone 15-20g / L and agar powder 15-20g / L, pH is natural, put into test tubes, stopper, extinguish at 115°C After sterilizing for 30 minutes, make a test tube slope (18mm×180mm), and set aside;

[0032] Preparation of solid plate medium: the formula is the same as that of the slant medium, sterilized at 115°C for 30 minutes, cooled to about 50°C, poured into a sterilized plate, and set aside.

[0033] Thaw the Aureobasidium pullulans AS3.3984 milk freezing tube, transfer it to the slant medium, and culture it in a constant temperature incubator at 28°C for 2 days; transfer it to a plate, and culture it in a constant temperature incubator at 28°C for 3 days, until a single colony grows , after 3 days of constant temperature culture at 28°C on a solid plate, ...

Embodiment 3

[0055] Aureobasidium pullulans strain (Aurebasidium pullulans) NCPS2016-M fermentation production pullulan 10L fermenter small test method, comprises the following steps:

[0056] Preparation of seed medium: glucose 15-20g / L, yeast extract powder 2-2.5g / L, ammonium sulfate 0.6-0.8g / L, magnesium sulfate heptahydrate 0.2-0.25g / L, sodium chloride 1.0-3.0g / L L and dipotassium hydrogen phosphate trihydrate 5.0-6.6g / L, pH 6.5, sterilized at 115°C for 30min;

[0057] Preparation of culture medium for upper tank: Calculated according to the liquid volume of 75%, 7.5L should be prepared: 750g of glucose, 940mL of 8.8% soybean meal acid hydrolyzate, (NH 4 ) 2 SO 4 4.5g, MgSO 4 ·7H 2 O 1.5g, NaCl 7.5g, K 2 HPO 4 ·3H 2 O 49.5g; pH 6.5, sterilized at 121°C for 20min, cooled.

[0058] Prepare 8.8% soybean meal acid hydrolysis solution: press 160g of soybean meal at low temperature, add 312.5mL of 1M hydrochloric acid, add distilled water to make up to 1250mL, acidify at 121°C for 3...

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Abstract

The invention belongs to the technical field of microbes and in particular relates to an aureobasidium pullulans stain NCPS 2016-M capable of producing pulullan at high yield. The aureobasidium pullulans stain NCPS 2016-M capable of producing pulullan at high yield provided by the invention is characterized in that the strain is collected in China Center for Type Culture Collection Center on September 14, 2017, and has a collection number of CCTCC M 2017498. The aureobasidium pullulans stain disclosed by the invention has the beneficial effects that the amount of the pulullan produced by performing shaking flask fermentation on the aureobasidium pullulans stain provided by the invention reaches 60g/L, and the ability of producing the pullulans is 1.5-1.7 times that of a wild aureobasidiumpullulans stain. The mutant strain does not produce any melanin, and the fermentation time is shortened by six hours compared with that of an original strain. The technical problem that wide application of the pullulans in the domestic food industry, particularly in an agricultural product preservation aspect, is greatly limited because the pullulans production level is low and the product qualityis low is solved.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of Aureobasidium pullulans with high yield of pullulan. Background technique [0002] Pullulan polysaccharide is favored by food, medicine and other industries for its excellent film-forming properties, gas barrier properties, water solubility, biocompatibility and degradability, especially in the preservation of agricultural products. focus on. As an excellent food additive and antistaling agent, because foreign products have been in a monopoly position, the price has remained high, and the selling price has been around 25 US dollars / kg. At present, the domestic production level of this product is low, and the product quality is not high, which greatly limits the wide application of pullulan in the domestic food industry, especially in the preservation of agricultural products. At present, Aureobasidium pullulans is considered to be an excellent str...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/10C12R1/645
CPCC12N1/14C12P19/10C12N1/145C12R2001/645
Inventor 杨金玉赵双枝陈蕾蕾张彦昊王易芬孙华张翔
Owner INST OF AGRO FOOD SCI & TECH SHANDONG ACAD OF AGRI SCI
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