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Mutant strain for mass production of pulullan and cultural method thereof

The technology of pullulan polysaccharide and shaking culture is applied in the field of mutagenic strain Aureobasidium pullulans and cultivation, which can solve the problems of low yield of pullulan polysaccharide, low pigment level, etc. The effect of improving the utilization rate of raw materials

Active Publication Date: 2013-04-24
天津北洋百川生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fang Xuanjun also obtained the mutant strain ZY047 through ultraviolet irradiation, and the polysaccharide conversion rate reached 54.1% by optimizing the culture conditions, and the pigment level was low; the title of Liu Na's 2005 master's thesis was "Composite mutagenesis screening of pullulan-producing bacteria and research on fermentation conditions" , this paper mainly studies the optimization of pullulan production strains and fermentation conditions, and determines the strains and fermentation conditions. Although its pigment level is low, its pullulan production is not high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 The original bacterial strain Aureobasidium pullulans TKPM10017 producing pullulan

[0019] 1. Cell morphology: Aureobasidium pullulans has a complex life cycle. During its growth, the cell morphology has the following five changes: yeast-like, spore-like, swollen spore, mycelium-like and chlamydospore. The evolution depends on the growth conditions. It has been found through research that the yield of polysaccharides is positively correlated with the number of yeast-like cells, and increases with the increase of the proportion of yeast-like cells.

[0020] 2. Colony morphology: Aureobasidium exhibits typical fungal growth, forming fungal colonies with loose edges and irregular edges; the colonies are dirty white and sticky at the beginning, and then gradually darken to black, leathery and wrinkled.

[0021] 3. Culture characteristics: This bacterium is a highly aerobic bacterium, the optimum growth temperature is 24-34 ℃, the slightly acidic environment is g...

Embodiment 2

[0024] Example 2 Taking Aureobasidium pullulans TKPM10017 as the starting strain to obtain bacterial strain BCSWGHPL101 by compound mutagenesis technique screening

[0025] 1. Obtaining of mutagenized strains

[0026] (1) Select the well-growing Aureobasidium pullulans strain TKPM10017 slant strain, wash the spores with sterile saline, and make 10 6 Individual / mL spore suspension, under 30W ultraviolet lamp, irradiate at 30cm for 0.5min, 1min, 1.5min, 2min, 2.5min3min, 3.5min, 4min respectively, dilute the spore suspension and spread it on a plate, and incubate at 28℃ in the dark , to calculate the fatality rate;

[0027] (2) Three different irradiation times of 0.5min, 2min, and 4min were selected to irradiate the spore suspension of strain TKPM10017 with ultraviolet light;

[0028] (3) Then mix the spore suspensions with three different irradiation times evenly, and perform 10-fold serial dilution 10 -1 ~10 -6 , take 10 -4 、10 -5 、10 -6 Three dilutions of the spore su...

Embodiment 3

[0042] The cultivation of embodiment 3 Aureobasidium pullulans BCSWGHPL101

[0043] 1. Activation of strains

[0044] Aureobasidium pullulans (Aureobasidium pullulans) BCSWGHPL101 was transferred to PDA slant medium and incubated at 28°C for 3 days.

[0045] 2. Seed culture

[0046] The well-growth activated slant strains were selected and transferred to the Erlenmeyer flask containing the seed medium, and cultured at 28°C, 200±20rpm for 36h.

[0047] 3. Fermentation culture

[0048] The seed medium prepared in 2 was inoculated into the fermentation medium at an inoculum amount of 5% (v / v), and cultured at 28°C, 200±20rpm for 3 days.

[0049] Test results: Aureobasidium pullulans (Aureobasidium pullulans) BCSWGHPL101 fermented in the fermentation medium to produce pullulan yield was 96g / L. The yield of the original strain was 68±5.2g / L, which was 41.2% higher than that of the original strain.

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Abstract

The invention discloses an aureobasidium pullulan for producing pullulan, and belongs to the field of microorganisms. The preservation number of the aureobasidium pullulan BCSWGHPL101 (Aureobsidium pullulans) is CGMCCNO.7055; the fermentation medium adopted by the aureobasidium pullulan for producing the pullulan comprises peptone of 3g / L, cane sugar of 100g / L, K2HPO4.7H2O of 3.0g / L, MgSO4.7H2O of 0.4g / L, NaCl of 2.5g / L and FeSO of 40.01g / L, and the pH is adjusted to 7.0. Compared with an ordinary pullulan producing strain, the mutant strain BCSWGHPL101 is higher in pullulan production capacity, and as a result, the yield of the pullulan is effectively increased, and the production cost is lowered.

Description

technical field [0001] The invention belongs to the field of fermentation engineering, and in particular relates to a mutagenic strain Aureobasidium pullulans capable of producing a large amount of pullulan and a cultivation method thereof. Background technique [0002] Pullulan is an exopolysaccharide produced by Aureobsidium 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 also has Non-toxic, harmless, colorless and tasteless and other excellent properties, it has been widely used in the fields of medicine, food, light industry, chemical industry and petroleum, and is a promising industrial polysaccharide. [0003] The cell morphology of Aureobsidium pullulans is various. During the fermentation process, due to the different fermentation conditions and substrates, there are generally two states of yeast (Y phase) and mycelium (M phase) that change ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/10C12R1/645
Inventor 乔长晟郝华璇卢星达李政马正旺
Owner 天津北洋百川生物技术有限公司
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