Aureobasidium pullulans for highly producing pigment-free Pulullan and method for culturing and preparing Pulullan

A pullulan polysaccharide and a culturing method are applied in the fields of Aureobasidium pullulans producing high-yield non-pigmented pullulan polysaccharide and its cultivation and preparation of pullulan polysaccharide, so as to achieve reduced production cost, high yield and raw material utilization rate. improved effect

Active Publication Date: 2014-06-25
INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is that the pullulan polysaccharide obtained by the existing production method is black or the pigment is lighter and the yield is generally not high. Ultraviolet radiation and nitrosoguanidine NTG mutagenesis, screening the mutagenic strains that are colorless, transparent and / or white in color and have a high transformation rate from the mutagenized strains, and obtain a strain that produces a large amount of pigment-free pullulan

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Screening of Aureobasidium pullulans mutant strains and their properties by compound mutagenesis treatment

[0028] (1) strain

[0029] The starting strain was Aureobasidium pullulans ATCC 201253, and the mutant strain after compound mutagenesis was Aureobasidium pullulans GM-1. The mutant strain Aureobasidium pullulans mutant strain GM-1 was deposited in the China Center for Type Culture Collection (CCTCC) on November 27, 2013, with the preservation number CCTCC M 2013611.

[0030] (2) Strain activation and seed cultivation

[0031] Transfer the starting strain Aureobasidium pullulans ATCC 201253 to PDA slant medium, culture at a constant temperature of 28-33°C for 3-4 days, and select the well-growing activated slant strains to transfer to the Erlenmeyer flask containing the seed medium cultured at 28-33°C for 36h at 200±20rpm.

[0032] (3) Strain selection and breeding

[0033] Taking Aureobasidium pullulans as the starting strain to carry out ultrav...

Embodiment 2

[0045] The cultivation and performance of embodiment 2 Aureobasidium pullulans mutant strain

[0046] Culture method: 1) Transfer the slant strain Aureobasidium pullulans mutant strain GM-1 to the slant PDA medium, activate and culture at 30°C for 3 days;

[0047] 2) Inoculate Aureobasidium pullulans GM-1 into a seed containing 20.0 g / L sucrose, 2.0 g / L peptone, 3.0 g / L rice bran, 0.3 g / L oxalic acid, K 2 HPO 4 1.0 g / L, NaCl 1.0 g / L, MgSO 4 ·7H 2 O 0.2 g / L, FeSO 4 ·7H 2 O 0.01 g / L, pH 6.0 liquid nutrient medium, cultured at 28°C with shaking at 220 rpm, and cultured to the stable period of proliferation (144 hours from inoculation). The culture obtained from the fermentation of the mutagenized mutant strain GM-1 is light milky white or light milk yellow, while the fermentation culture of the starting parent strain Aureobasidium pullulans ATCC 201253 is dark green or dark green.

[0048] In order to measure the glucan content in the aforementioned culture, collect the ...

Embodiment 3

[0058] Example 3 Pigmentless fermentation production method of pullulan

[0059] Utilize the optimized medium formula, and at the same time, by controlling the fermentation conditions, first promote the growth of bacteria, and then change the fermentation conditions to promote product synthesis, realize the fermentation and production of pullulan without pigment, make full use of the medium substrate, and improve the yield of raw materials The utilization rate reduces the production cost.

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Abstract

The invention discloses an aureobasidium pullulans for highly producing pigment-free Pulullan, which is collected in China Center For Type Culture Collection (CCTCC) with a collection number of CCTCC M2013611. The invention further discloses a mutation screening method and a culture method of aureobasidium pullulans for highly producing pigment-free Pulullan, and a method for producing Pulullan through fermentation of the aureobasidium pullulans for highly producing pigment-free Pulullan. A culture obtained by fermentation cultivation of an aureobasidium pullulans strain GM-1 cannot show a dark green color or deep green color due to accumulation of melanin substance, but form a milk white or faint yellow composition. Compared with a conventional Pullulan producing strain, the aureobasidium pullulans strain GM-1 has the advantages that the Pulullan is colorless and is higher in yield, the purposes of increasing the raw material utilization rate and reducing the production cost are reached, and the demand of application fields such as foods, medicines, and environment on the pigment-free Pulullan is met.

Description

technical field [0001] The invention belongs to the technical field of microbial strain bioengineering of pullulan polysaccharide, and in particular relates to a mutagenic Aureobasidium pullulan strain and a fermentation culture method for producing a large amount of pigment-free pullulan polysaccharide. [0002] Aureobasidium pullulans of the present invention is preserved in CCTCC, China's Typical Culture Collection Center, the preservation address is Wuhan University, Wuhan, China, the preservation date is November 27, 2013, and the preservation number is CCTCC M 2013611; preservation name: Aureobasidium pullulans GM -1 (Aureobasidium pullulans GM-1). [0003] technical background [0004] Pullulan is an exopolysaccharide produced by Aureobasidium pullulans. It is easily soluble in water and has a very good thickening effect. It has strong film-forming properties and is easy to form a good water-soluble edible film. It has excellent characteristics such as non-toxic, har...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N15/01C12P19/10C12R1/645
Inventor 袁林郭建军张小华杨罡
Owner INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI
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