Aureobasidium Pullulan producing pullulan with high yield by utilizing xylose and application of Aureobasidium Pullulan

A technology of Aureobasidium pullulans and pullulan, which is applied in the direction of microorganism-based methods, microorganisms, fungi, etc., can solve the problems of complex process and high cost, and achieve the effect of increasing added value

Active Publication Date: 2013-08-21
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Use physical methods to separate the components in the xylose mot...

Method used

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  • Aureobasidium Pullulan producing pullulan with high yield by utilizing xylose and application of Aureobasidium Pullulan
  • Aureobasidium Pullulan producing pullulan with high yield by utilizing xylose and application of Aureobasidium Pullulan
  • Aureobasidium Pullulan producing pullulan with high yield by utilizing xylose and application of Aureobasidium Pullulan

Examples

Experimental program
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Effect test

Embodiment 1

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

[0026] Take Aureobasidium pullulan (Aureobasidium Pullulan) CGMCC3.00837 as the starting strain, select the ultraviolet mutagenic dose (irradiation 5min) with a lethality rate of 82.4% to carry out mutagenesis to Aureobasidium pullulan, dilute the bacterium solution after the mutation and evenly Spread on a high-concentration xylose plate (xylose concentration 100g / L), culture at 30°C in the dark 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.

[0...

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.

[0037] 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.

[0038] Table 3 AY82 xylose mother liquor medium fermentation result

[0039]

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Abstract

The invention discloses an Aureobasidium Pullulan producing pullulan with high yield by utilizing xylose and a production method of pullulan. The Aureobasidium Pullulan is taken as an original strain and subjected to ultraviolet mutagenesis to obtain a mutated strain which produces pullulan with high yield and secretes less pigment. The Aureobasidium Pullulan mutated strain AY82 is preserved in the China General Microbiological Culture Center with the data of Jan 18th, 2013 and the preservation number of CGMCC No.7154. The Aureobasidium Pullulan mutated strain AY82 can produce pullulan by utilizing fermentation of xylose, yield of pullulan is higher, after shake-flask culture is carried out on xylose fermentation culture medium for seven days, the yield of pullulan is 16.61g/L, and after culture by virtue of a 5L fermentation tank is carried out for seven days, the yield is 16.23g/L. Study on the Aureobasidium Pullulan producing pullulan by utilizing xylose can provide a new method for reasonably utilizing xylose, and additional value of xylose can be effectively enhanced.

Description

Technical field: [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a strain of Aureobasidium pullulans using xylose to produce pullulan and its application. 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 micro...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/10C12R1/645
Inventor 陈叶福肖冬光郭建郭学武张翠英董健杜丽平
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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