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Method for increasing pulullan output through fed-batch fermentation

A pullulan polysaccharide and fed-feed fermentation technology, which is applied in the field of fed-fed fermentation to increase the yield of pullulan polysaccharide, can solve the problems of unfavorable large-scale, large-scale production, high cost of pullulan polysaccharide, and low substrate utilization rate , to achieve the effect of improving substrate conversion rate, convenient operation and shortening fermentation cycle

Inactive Publication Date: 2015-09-16
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

However, it still has the problems of long fermentation cycle, low substrate conversion rate and low substrate utilization rate, resulting in high cost of pullulan polysaccharide, which is not conducive to large-scale and large-scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1) Seed culture

[0041] After activating the strain of Aureobasidium pullulans, transfer it to a 500ml baffle bottle, the volume of the culture medium is 100ml, the culture temperature is 32°C, the shaker speed is 180rpm, and the culture time is 28h;

[0042] 2) Fermentation culture

[0043] The 5L fermentation tank has a liquid volume of 1.8L, an inoculum size of 4% (V / V), and a culture temperature of 30°C. After 24 hours of fermentation, feeding solution is started to control the residual sugar of the fermentation liquid at 6-12g / L. Add 0.8g / L of phenylalanine solution once every two hours, and the fermentation time is 67h;

[0044] 3) Separation of pullulan

[0045] After filtering out the bacterial cells in the fermentation broth, add twice as much ethanol, stir, let stand overnight at 4°C, centrifuge, rinse twice with absolute ethanol, and dry to obtain 74.6 g / L of white pullulan crude product. The fermentation period was 67h, which was shortened by 7h compared...

Embodiment 2

[0047] 1) Seed culture

[0048] After activating the strain of Aureobasidium pullulans, transfer it to a 500ml baffle bottle, the volume of the culture medium is 100ml, the culture temperature is 32°C, the shaker speed is 180rpm, and the culture time is 30h;

[0049] 2) Fermentation culture

[0050] The 5L fermenter has a liquid volume of 1.8L, an inoculum size of 4% (V / V), a culture temperature of 30°C, and after 28 hours of fermentation, feeding solution is started to control the residual sugar of the fermentation liquid at 6-12g / L. Add an arginine solution of 1.5g / L every two hours, and the fermentation time is 65h;

[0051] 3) Separation of pullulan

[0052] After the bacterial cells were filtered out from the fermentation broth, twice the amount of ethanol was added, stirred, left to stand overnight at 4°C, centrifuged, rinsed twice with absolute ethanol, and dried to obtain 73.9 g / L of crude white pullulan. The fermentation period was 65h; compared with the fermentati...

Embodiment 3

[0054] 1) After activating the strain of Aureobasidium pullulans, transfer it to a 500ml baffle bottle, the volume of the culture medium is 100ml, the culture temperature is 32°C, the rotation speed of the shaker is 180rpm, and the culture time is 30h;

[0055] 2) Fermentation culture

[0056] The 5L fermentation tank has a liquid volume of 1.8L, an inoculum size of 4% (V / V), a culture temperature of 30°C, and feed feeding solution after 25 hours of fermentation to control the residual sugar of the fermentation liquid at 6-12g / L. Add 1.3g / L lysine solution and 1.8g / L histidine solution once every two hours, and ferment for 66 hours;

[0057] 3) Separation of pullulan

[0058] After filtering out the bacterial cells in the fermentation broth, add twice as much ethanol, stir, let stand overnight at 4°C, centrifuge, rinse twice with absolute ethanol, and dry to obtain 73.3 g / L of white pullulan crude product. The fermentation period was 66h, which was shortened by 8h compared wit...

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Abstract

The invention relates to a method for increasing pulullan output through fed-batch fermentation, which comprises the following steps: culturing seeds, performing fermentation culture and separating pulullan; continuously feeding a fed-batch solution after fermenting for 24-28 hours, controlling the residual sugar of a broth to maintain at 6-12g / L, supplementing 0.5-2g / L of amino acid solution for once by every two hours, wherein the total fermentation period is 65-67 hours. Aiming at defects of long period and low substrate conversion rate during a process of fermentation of aureobasidium pullulans with a preservation number of No. 7055 to produce pulullan, a proper amount of amino acid is continuously fed during the fermentation process for synthesis of pullulan, substrate conversion rate is increased, operation is convenient, effect is obvious, fermentation period is greatly shortened, and cost of pullulan can be reduced.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a method for increasing the yield of pullulan through fed-batch fermentation. Background technique [0002] Pullulan is a kind of extracellular water-soluble mucilaginous microbial exopolysaccharide similar to dextran and xanthan gum. It is formed by combining glucose with α-1,4-glycosidic bonds into maltotriose, and the two ends are connected with α- The 1,6-glucosidic bond is combined with another maltotriose, so that it is repeatedly connected to form a high molecular polymer. Pullulan has been used as an emulsifier, suspending agent, thickener, stabilizer, gelling agent, film-forming agent and lubricant, and has been widely used in food, pharmaceutical, petroleum, chemical and other fields. It will be an important development direction of new fermentation engineering in the future, and it will be more and more valued by domestic enterprises. [0003] In the c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/10C12R1/645
Inventor 乔长晟王建梓郝华璇李振海
Owner 天津北洋百川生物技术有限公司
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