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Method for producing pullulan by aureobasidium pullulans AY82 by using xylose

A technology of Aureobasidium pullulans and pullulan sugar, applied in the field of microbial fermentation, can solve the problems of complex process and high cost, and achieve the effect of increasing added value

Active Publication Date: 2014-09-10
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Use physical methods to separate the components in the xylose mother liquor and use them, the process is complicated and the cost is high

Method used

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  • Method for producing pullulan by aureobasidium pullulans AY82 by using xylose
  • Method for producing pullulan by aureobasidium pullulans AY82 by using xylose
  • Method for producing pullulan by aureobasidium pullulans AY82 by using xylose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Screening of mutant strains of Aureobasidium pullulans with high yield of pullulan using xylose

[0026] Taking Aureobasidium pullulan (Aureobasidium Pullulan) CGMCC 3.00837 as the starting strain, the ultraviolet mutagenic dose (irradiation 5min) with a lethality rate of 82.4% was selected to mutate Aureobasidium pullulan. Spread on high-concentration xylose plates (xylose concentration 100g / L), culture in the dark at 30°C for 4-5 days, and then pick mutants. The mutant strains were re-screened in the shake flask fermentation experiment, and the amount of pullulan sugar produced was measured after 168 hours of fermentation. Compared with the original strain, a mutant strain AY82 that utilizes xylose to produce pullulan sugar was obtained, and the pullulan sugar yield was relatively high. The starting strain has been greatly improved, and the fermentation results are shown in Table 1.

[0027] Table 1 Comparison of pullulan production between the mutant stra...

Embodiment 2

[0029] Embodiment 2: Shake flask fermentation of xylose medium

[0030] Taking the mutant strain AY82 as the experimental strain, using xylose as the carbon source, and optimizing the medium and fermentation conditions, the optimized xylose fermentation medium is: xylose 50g / L, (NH 4 ) 2 SO 4 1.2g / L, K 2 HPO 4 6g / L, MgSO 4 ·7H 2 O0.4g / L, NaCl2g / L, yeast extract powder 1.0g / L, sterilized by high-pressure steam at 121°C for 20min; the optimized fermentation conditions are: inoculum size 8%, shaker speed 200r / min, temperature 28°C, initial pH The value is 6.5, and the culture time is 168h.

[0031] Pick a loop of AY82 bacteria from the activated slope and inoculate it into a 250mL Erlenmeyer flask containing 50mL of seed medium for cultivation, and cultivate it on a shaker at 28°C and 200r / min for 24-48h to obtain a seed solution. The composition of the seed medium is: xylose 20g / L, (NH 4 ) 2 SO 4 1.0g / L, K 2 HPO 4 4g / L, MgSO 4 ·7H 2 O0.2g / L, NaCl4g / L, yeast extract...

Embodiment 3

[0035] Embodiment 3: Shake flask fermentation of xylose mother liquor culture medium

[0036]The preparation of the seed liquid and the shake flask fermentation conditions were basically the same as in Example 2. The components of the fermentation medium are: xylose mother liquor (xylose 35-40%, arabinose 8-16%, galactose 9-15%, glucose 8-11%) diluted to a total sugar content of 50g / L in the medium , (NH 4 ) 2 SO 4 1.2g / L, K 2 HPO 4 6g / L, MgSO 4 ·7H 2 O0.4g / L, NaCl2g / L, yeast extract powder 1.0g / L, initial pH6.5. Sterilized by high pressure steam at 121°C for 20min. The fermentation results are shown in Table 3.

[0037] Table 3 AY82 xylose mother liquor medium fermentation result

[0038]

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Abstract

The invention discloses a method for producing pullulan by aureobasidium pullulans AY82 by using xylose. A mutant strain which has high yield of the pullulan and secretes little pigment is obtained by performing ultraviolet mutagenesis on the aureobasidium pullulans serving as an original strain. The aureobasidium pullulans mutant strain AY82 is preserved in the China general microbiological culture collection center (CGMCC) in January, 18th in 2013 with the preservation number of CGMCC No.7154. The strain can produce the pullulan by using the xylose by fermentation; the yield of the pullulan is higher; after a xylose fermentation culture medium shake flask is cultivated for seven days, the yield of the pullulan is up to 16.61 g / L; and after a fermentation pot of 5 liters is cultivated for seven days, the yield is up to 16.23 g / L. A research shows that the pullulan can be produced by the aureobasidium pullulans by using the xylose; a new concept can be supplied to rational utilization of the xylose; and the additional value of the xylose can be effectively increased.

Description

Technical field: [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to an Aureobasidium pullulans strain and a method for producing pullulan by using xylose. Background technique: [0002] Pullulan is an unbranched extracellular homopolysaccharide produced by Aureobasidium Pullulan, which is composed of α-(1,4) and α-(1,6) glycosidic bonds. Sugar is continuously polymerized. The regular chemical structure endows pullulan with unique properties. Its structure is flexible and easily soluble in water. Pullulan can be used as a thickener, and a small amount of pullulan dissolved in water can produce a large viscosity. The transparent film made of pullulan is oil-proof and impermeable to oxygen. Its good performance makes it suitable for coating and degradable packaging materials of food and medicine. Pullulan can also replace starch as a low-calorie food, and can also be used as a cosmetic emulsion. Pullulan is a microbia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/10C12R1/645
Inventor 陈叶福肖冬光郭建郭学武张翠英董健杜丽平
Owner TIANJIN UNIV OF SCI & TECH
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