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Aureobasidium pullulans and application thereof

A technology of Aureobasidium pullulans and strains, applied in the direction of microorganisms, fungi, biochemical equipment and methods, etc., can solve the problems affecting the yield and molecular weight of pullulan, the reduction of synthetase activity, and the inhibition of cell growth, etc., and achieve good results. Industrial application prospects, short fermentation time, and good social and economic benefits

Inactive Publication Date: 2013-03-27
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Acid stress can lead to inhibition of cell growth and decrease of pullulan synthetase activity, thus affecting the yield and molecular weight of pullulan

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Screening of Aureobasidium pullulans mutant strains and performance comparison by compound mutagenesis

[0032] (1) strain

[0033] The starting strain was Aureobasidium pullulans ( Aureobasidium pullulans ATCC 201253), the mutant strain after compound mutagenesis is Aureobasidium pullulans SZU 1001. The mutant strain was deposited in China Center for Type Culture Collection (CCTCC) on June 29, 2012, with the preservation number CCTCC M 2012259.

[0034] (2) Medium

[0035] Seed medium: PDA medium (potato extract 1 L, glucose 20 g, natural pH);

[0036] Screening medium: add 0.1 g / L ethionine to the seed medium;

[0037] Fermentation medium before optimization: 50 g / L glucose, 2.5 g / L yeast powder, 5 g / L K 2 HPO 4 , 0.6 g / L (NH 4 ) 2 SO 4 , 0.2 g / L MgSO 4 ·7H 2 O, 1 g / L NaCl, pH 6.8.

[0038] (3) UV mutagenesis

[0039]Irradiation was carried out under the Philips ZWSSJD-30 ultraviolet lamp, the irradiation power and wavelength were 30W, 253....

Embodiment 2

[0061] Example 2: Determining the best medium for high-yield fermentation of high-molecular-weight pullulan

[0062] The operation was carried out as in Example 1, with the difference that: taking the fermentation medium before optimization as the benchmark, the fermentation medium was optimized by using an orthogonal design test to determine the optimum medium composition and concentration.

[0063] According to the results of the orthogonal design experiment, the most important nutrients affecting the yield of pullulan are ammonium sulfate, yeast powder, sodium chloride and dipotassium hydrogen phosphate in sequence, and the optimal concentrations are: ammonium sulfate 0.8 g / L, yeast powder 3 g / L, sodium chloride 1 g / L, dipotassium hydrogen phosphate 3 g / L.

[0064] According to the results of orthogonal design experiments, the most important nutrients that affect the molecular weight of pullulan are sodium chloride, yeast powder, dipotassium hydrogen phosphate and ammonium ...

Embodiment 3

[0065] Example 3: The optimized medium (1) was used for the pullulan fermentation culture of mutant strains.

[0066] Each liter of fermentation medium (1) contains: 50 grams of glucose, 3.0 grams of yeast powder, 0.8 grams of ammonium sulfate, 3.0 grams of dipotassium hydrogen phosphate, 0.2 grams of magnesium sulfate, 1 gram of sodium chloride, the pH of the fermentation medium It is 6.8.

[0067] Carry out seed culture and fermentation culture with embodiment one, the result is as follows:

[0068] Cell dry weight 9.9 g / L; pullulan yield 22.3 g / L; pullulan molecular weight 1.69×10 6 .

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Abstract

The invention discloses Aureobasidium pullulans and application thereof. The Aureobasidium pullulans strain SZU1001 is preserved in China center for type culture collection (CCTCC), and the preservation number is CCTCC M 2012259. A method for producing pullulan by the Aureobasidium pullulans strain includes the steps of by using the Aureobasidium pullulans strain as an original strain, performing fermentation cultivation for at least 48 hours after slope seed activation and seed culture, and extracting with an organic solvent to obtain the pullulan. The method is characterized in that the fermentation medium per liter contains glucose 40-60g, yeast powder 0.8-3.5g, ammonium sulfate 0.2-1.0g, dipotassium phosphate 1.0-3.0g, magnesium sulfate 0.1-0.3g, and sodium chloride 0-1.0g, and the pH value is 6.5-6.8. According to the invention, the aureobasidium pullulans mutant strain which can synthesize glutathione in cells can be obtained, so that the synthesis of pullulan with high molecular weight and no pigment production is realized, thereby meeting the requirement of application fields, such as medicine and environment, on the pullulan.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a strain of Aureobasidium pullulans which does not produce pigment and has the ability to synthesize glutathione and its application in the synthesis of high-molecular-weight pullulan. Background technique [0002] Pullulan (Pullulan), also known as pullulan, is an aureobasidium pullulan ( Aureobasidium pullulans ) The exopolysaccharide produced is a high molecular polysaccharide composed of α-1,6-glycosidic bonds combined with maltotriose. The pullulan molecule is a linear structure with no branches, and its molecular weight is generally 4.8×10 4 ?2.2×10 6 Between (the average molecular weight of commercial pullulan polysaccharide is 2×10 5 , composed of approximately 480 maltotriose), the degree of polymerization is 100-5000, and its optical rotation is 192° at a concentration of 10 g / L. [0003] Pullulan is a colorless, tasteless, and odorless po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P19/04C12R1/645
Inventor 卫功元余小六王大慧
Owner SUZHOU UNIV
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