Method for increasing output of alkaline pectinase produced by fermentation by adding sorbitol

The technology of sorbitol and pectinase is applied in the application field of carbon source addition strategy in the optimization of fermentation process, so as to achieve the effects of increasing yield, weakening the toxic effect of methanol and improving the efficiency of enzyme production

Inactive Publication Date: 2009-09-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The low-speed fed-batch of sorbitol can solve this problem and improve the yield of pectinase, which has not been reported in the literature yet.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Fermentation in a fermenter adopts the strategy of adding sorbitol at a constant rate, and the rest of the conditions are the same as those in the comparative example. Sorbitol (50%, v / v, 1.8 g / h solid sorbitol) was added when the fermentation entered the induction phase. Results: The production of alkaline pectinase increased from 930U / mL of the control to 1176U / mL.

Embodiment 2

[0020] Example 2: Fermentation in a fermenter adopts a strategy of adding sorbitol at a constant rate, and the rest of the conditions are the same as those in the comparative example. Sorbitol (50%, v / v, 3.6 g / h solid sorbitol) was added when the fermentation entered the induction phase. Results: The production of alkaline pectinase increased from 930U / mL of the control to 1389U / mL.

Embodiment 3

[0021] Example 3: Fermentation in a fermenter adopts the strategy of adding sorbitol at a constant rate, and the rest of the conditions are the same as those in the comparative example. Sorbitol (50%, v / v, 5.4 g / h solid sorbitol) was added when the fermentation entered the induction phase. Results: The production of alkaline pectinase increased from 930U / mL of the control to 1211U / mL.

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Abstract

The invention discloses a method for increasing the output of alkaline pectinase produced by fermentation by adding sorbitol, relating to the technical field of application of the carbon source adding policy in the optimization of fermentation. In the method, recombinant Pichia pastoris GS115 is adopted as fermentation strains, and sorbitol is added into the fermentation liquid at a constant speed or accelerated speed during fermentation induction phase to improve the activity of cells, weaken the toxicity of methanol, and increase the output of the alkaline pectinase. The lower adding amount of sorbitol can not inhibit the enzyme production effect of methanol to the strains, on the other hand, the toxicity to the strains from the methanol can be reduced to a certain extent, and the efficiency of enzyme production is further increased. When the sorbitol is added at a varying speed (0.9g / h-7.2g / h), the activity of enzyme is increased from 930U / ml to 1541U / ml. The invention is simple and effective in process, capable of being applied in the industrial production of pectinase in the future, and instructive to the optimization of fermentation of other Pichia pastoris.

Description

technical field [0001] Using recombinant Pichia pastoris as a host to efficiently express alkaline pectinase, a method for increasing the yield of alkaline pectinase by adding sorbitol, relates to the technical field of application of carbon source addition strategy in fermentation process optimization. Background technique [0002] Alkaline pectinase is a kind of enzyme that can efficiently decompose pectin (a linear polymer formed by D-galacturonic acid linked by α-1,4 glycosidic bonds) in plant tissue under alkaline conditions As a general term for a new mild bioscouring enzyme preparation for textile cleaning production, it has attracted much attention. In the previous research, on the basis of isolation and screening of an alkaline pectinase high-yielding strain WSHB04-02, the prokaryotic alkaline pectin ester lyase gene PL was connected with the shuttle vector pPIC9K and successfully expressed In Pichia pastoris GS115, the strain preservation number is CGMCC NO.2143. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/26C12R1/84
Inventor 陈坚汪志浩李江华堵国成张东旭
Owner JIANGNAN UNIV
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