Fermentation method for producing pullulan polysaccharide

A technology of pullulan polysaccharide and fermentation method, which is applied in the field of microbial fermentation, can solve the problems of long fermentation time and low yield, and achieve the effects of shortening fermentation time, increasing product yield, and small molecular weight range

Active Publication Date: 2008-07-09
天津量信检验认证技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long fermentation time, low yield and wide range of molecular weight distr

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] (1) Take a ring of Aureobasidium pullulans and insert it into a 500 ml Erlenmeyer flask with 100 ml of seed culture medium. Shake flask seed culture medium, its aqueous solution is composed of (g / L): soybean oil 30.0, KH 2 PO 4 6.0, (NH 4 ) 2 SO 4 0.5, MgSO 4 .7H 2 O0.2, NaCl1.0, FeSO 4 .7H 2 O0.01, yeast extract 0.5, pH value 6.0, sterilized at 121°C for 20 minutes, shake the flask at 28°C, and cultivate it on a shaker with a rotation speed of 240rpm for 48 hours, as the primary seed solution.

[0016] (2) Take 5 milliliters of the primary seed solution and put it into a 500 milliliter Erlenmeyer flask containing 100 milliliters of seed medium. The composition of the shake flask seed medium is the same as that in step 1. Sterilize at 121°C for 20 minutes, and culture the shake flask at 28°C on a shaker with a rotation speed of 240rpm for 24 hours as the secondary seed solution.

[0017] (3) Put the shake flask seeds into a 30-liter fermenter according to the i...

Embodiment 2

[0021] (1) Take a ring of Aureobasidium pullulans and insert it into a 500 ml Erlenmeyer flask with 100 ml of seed culture medium. Shake flask seed medium, its aqueous solution is composed of (g / L): peanut oil 40.0, KH 2 PO 4 5.0, (NH 4 ) 2 SO 4 0.6, MgSO 4 .7H 2 O0.2, NaCl0.8, FeSO 4 .7H 2 O0.01, yeast extract 0.4, pH value 5.5, sterilized at 121°C for 20 minutes, shake the flask at 28°C, and cultivate it on a shaker with a rotation speed of 240rpm for 48 hours, as the primary seed solution.

[0022] (2) Take 5 milliliters of the primary seed solution and put it into a 500 milliliter Erlenmeyer flask containing 100 milliliters of seed medium. The composition of the shake flask seed medium is the same as 1. Sterilize at 121°C for 20 minutes, and culture the shake flask at 28°C on a shaker with a rotation speed of 240rpm for 24 hours as the secondary seed solution.

[0023](3) Put the shake flask seeds into a 30-liter fermenter according to the inoculum amount of 5%. ...

Embodiment 3

[0027] (1) Take a ring of Aureobasidium pullulans and insert it into a 500 ml Erlenmeyer flask with 100 ml of seed culture medium. Shake flask seed medium, its aqueous solution is composed of (g / L): soybean oil 40.0, KH 2 PO 4 6.0, (NH 4 ) 2 SO 4 0.5, MgSO 4 .7H 2 O0.2, NaCl1.0, FeSO 4 .7H 2 O0.01, yeast extract 0.6, pH value 5.5, sterilized at 121°C for 20 minutes, shake the flask at 28°C, and cultivate it on a shaking table with a rotation speed of 240rpm for 48 hours, as the primary seed solution.

[0028] (2) Take 5 milliliters of the primary seed solution and put it into a 500 milliliter Erlenmeyer flask containing 100 milliliters of seed medium. The composition of the shake flask seed medium is the same as 1. Sterilize at 121°C for 20 minutes, and culture the shake flask at 28°C on a shaker with a rotation speed of 240rpm for 24 hours as the secondary seed solution.

[0029] (3) Put the shake flask seeds into a 30-liter fermenter according to the inoculum amount...

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Abstract

The invention relates to a fermenting method for producing pullulanuse polysaccharide, which is characterized in that the method comprises following steps: firstly, preparing seed culture medium, secondly, preparing fermenting initial culture medium, thirdly, inoculating liquid seeds into the fermenting initial culture medium, wherein the inoculation amount is 3-8%, the fermentation stirring speed is 200-500rpm, the fermentation temperature is 29 DEG C +- 1 DEG C, the ventilation volume is 0.5-10V/V, and the bed pressure is 0.01-0.02MPa, fourthly, complementarily adding the other carbon source after fermenting for 24 hours, wherein the adding amount is 40-80g/L and the adding liuqid is sucrose solution or glucose solution or starch hydrolysate whose pH value is 40-60, and continuingly fermenting for 60-72 hours after adding the carbon source, fifthly, flocculating, hyper-filtrating, barrier-separating, concentrating and drying the fermenting liquid and obtaining the pullulanuse polysaccharide whose molecular weight is between 20-60 million daltons. The invention has simple technique and significant effects and prominently shortens the fermenting time, the fermenting time is 80-100 hours, the production efficiency is increased greatly, energy is saved, the yield rate of product is greatly improved, and the yield can reach by 50%.

Description

technical field [0001] The invention relates to a microbial fermentation method, in particular to a fermentation method for producing pullulan. Background technique [0002] Pullulan was discovered as early as 1936. Later, Aureobasidium pullulans were cultured in liquid and pullulan was isolated. It was not until the 1960s that people began to study its fermentation, extraction and application. In 1978, the Japanese Hayashibara Company realized the industrial production of pullulan. Pullulan is a viscous exopolysaccharide produced by aerobic fermentation of Aurebasidium pullulans using carbohydrate compounds as substrates. Pullulan is a linear polymer polysaccharide with maltotriose as the unit and repeatedly connected by α-1.6 bonds. Molecular weight 4.8×10 4 -2.2×10 6 , the average molecular weight of commercial pullulan polysaccharide is 2×10 5 . Pullulan is a non-ionic, non-reducing stable polysaccharide, so it is easy to dissolve in water and can be used as a visc...

Claims

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Application Information

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IPC IPC(8): C12P19/04C12R1/645
Inventor 许勤虎徐勇虎国华刘晓鸥李睿颖孙溪墨玉欣刘晨
Owner 天津量信检验认证技术有限公司
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