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Culture medium for promoting recombinant pichia pastoris to express xylanase and preparation method thereof

A technology for xylanase and Pichia pastoris, which is applied to a medium for promoting recombinant Pichia pastoris to express xylanase and the field of preparation thereof, can solve the problems of low initial pH, precipitation, expensive raw materials and the like, and achieves clear composition. , the effect of price economy

Inactive Publication Date: 2016-03-23
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a culture medium for promoting recombinant Pichia pastoris to express xylanase and a preparation method thereof, so as to solve the problem of complex components of the existing recombinant Pichia pastoris expressing xylanase, high raw material prices, The production conditions are not easy to control and the initial pH of the synthetic medium is low and precipitation is likely to occur, which affects the growth of cells and the expression of foreign proteins. A synthetic medium for basic research and commercial production of xylanase expressed by red yeast, and a method for preparing the medium

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The formulation of the recombinant Pichia pastoris GS115 expression xylanase induction medium of the present embodiment consists of: sodium glycerophosphate 29.0g, ammonium sulfate 4.5g, calcium sulfate dihydrate 1.6g, potassium sulfate 14.0g, magnesium sulfate heptahydrate 11.0g g, 2.4 mL of PTM1 trace element solution, 10.0 mL of anhydrous methanol, 802.0 g of Tween, dissolved in distilled water to 1000.0 mL, and adjusted the pH to 7.0 with KOH.

[0019] The preparation method of this culture medium is:

[0020] (1) Preparation of basic synthetic medium: Weigh 29.0 g of sodium glycerophosphate, 4.5 g of ammonium sulfate, 1.6 g of calcium sulfate dihydrate, 14.0 g of potassium sulfate, 11.0 g of magnesium sulfate heptahydrate, and 802.0 g of Tween. Distill water to 1000mL, adjust the pH to 7.0 with KOH, sterilize at 121°C for 20 minutes, and obtain the basic synthetic medium after cooling.

[0021] (2) Add trace element PTM1: 2.4mL PTM1 trace element is filtered and s...

Embodiment 2

[0025] The formula composition of recombinant Pichia pastoris GS115 expression xylanase induction medium is as follows: sodium glycerophosphate 28.0g, ammonium sulfate 3.5g, calcium sulfate dihydrate 2.5g, potassium sulfate 10.0g, magnesium sulfate heptahydrate 10.0g, PTM1 trace Element solution 4.8mL, anhydrous methanol 12.0mL, Tween 802.8g, distilled water dissolved to 1000.0mL, and KOH was used to adjust the pH to 7.5.

[0026] The preparation method of this culture medium is:

[0027] (1) Preparation of basic synthetic medium: Weigh 28.0g of sodium glycerophosphate, 3.5g of ammonium sulfate, 2.5g of calcium sulfate dihydrate, 10.0g of potassium sulfate, 10.0g of magnesium sulfate heptahydrate, and 802.8g of Tween. Distill water to 1000mL, adjust the pH to 7.5 with KOH, sterilize at 121°C for 20 minutes, and obtain the basic synthetic medium after cooling.

[0028] (2) Add trace element PTM1: 4.8mL PTM1 trace element is filtered and sterilized, and then aseptically added t...

Embodiment 3

[0032] The formulation of the recombinant Pichia pastoris GS115 expression xylanase induction medium consists of: sodium glycerophosphate 30.0g, ammonium sulfate 7.0g, calcium sulfate dihydrate 1.6g, potassium sulfate 14.0g, magnesium sulfate heptahydrate 11.0g, PTM1 trace Element solution 2.8mL, anhydrous methanol 10.0mL, Tween 802.3g, distilled water dissolved to 1000.0mL, and KOH was used to adjust the pH to 7.0.

[0033] The preparation method of this culture medium is:

[0034] (1) Preparation of basic synthetic medium: Weigh 30.0 g of sodium glycerophosphate, 7.0 g of ammonium sulfate, 1.6 g of calcium sulfate dihydrate, 14.0 g of potassium sulfate, 11.0 g of magnesium sulfate heptahydrate, and 802.3 g of Tween. Distill water to 1000mL, adjust the pH to 7.0 with KOH, sterilize at 121°C for 20 minutes, and obtain the basic synthetic medium after cooling.

[0035](2) Add trace element PTM1: 2.8mL PTM1 trace element is filtered and sterilized, and then aseptically added to...

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Abstract

The invention provides a culture medium for promoting recombinant pichia pastoris to express xylanase and a preparation method of the culture medium, wherein the culture medium is prepared from the following components: 15.0-40.0 g of sodium glycerophosphate, 1.0-10.0 g of ammonium sulfate, 1.0-4.0 g of calcium sulfate dihydrate, 8.0-16.0 g of potassium sulfate, 7.0-15.0 g of magnesium sulfate heptahydrate, 0.5-6.0 mL of PTM1 trace element solution, 5.0-20.0 mL of absolute methanol and 1.0-3.0 g of Tween80; the preparation method comprises the following steps of dissolving the components in distilled water to 1000 mL, and adjusting the pH to be 5.5-7.5 by utilization of KOH. The culture medium for promoting the recombinant pichia pastoris to express the xylanase and the preparation method of the culture medium have the advantages that the components are specific, the price is economic, the recombinant pichia pastoris well grows in the culture medium, and the culture medium is suitable for expressing the xylanase in an industrialized large-scale manner.

Description

technical field [0001] The invention belongs to the technical field of production of xylanase products, in particular to a culture medium for promoting the expression of xylanase by recombinant Pichia pastoris and a preparation method thereof. Background technique [0002] Pichia pastoris is one of the most widely used strains for recombinant protein production. It has the advantages of post-translational modification, relatively simple genetic operation, elimination of endotoxin and phage contamination, high expression level, secretable product, and high-density fermentation with simple inorganic salt medium. [0003] Xylanase is one of the commonly used industrial enzymes, and has important applications in the fields of medicine, health care products, food, feed and new energy. At present, xylanase is mainly expressed and produced by recombinant Pichia pastoris. [0004] At present, the recombinant Pichia pastoris induction medium commonly used in the laboratory is mainl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12R1/84
CPCC12N9/248C12N1/16
Inventor 朱新术赵文慧周晨妍罗晓秋解丽芹李同彪
Owner XINXIANG MEDICAL UNIV
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