Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Process for preparing phytase by efficiently fermenting recombinant pichiapastoris

A technology of Pichia pastoris and phytase, applied in the direction of hydrolase, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of high preparation cost of phytase, large amount of methanol added, etc., and achieve high-efficiency fermentation production Effect

Inactive Publication Date: 2010-11-24
WENZHOU UNIVERSITY
View PDF2 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the whole, there are defects in the salt composition in the medium and the addition of too much methanol in the induction stage, and the high cost of phytase preparation.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Process for preparing phytase by efficiently fermenting recombinant pichiapastoris

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The strain used was Pichia pastoris 602, provided by Wenzhou Conch Challenge Bioengineering Co., Ltd.

[0021] Pick a single colony from the plate and insert it into YPD seed medium (500mL Erlenmeyer flask liquid volume: 125mL) (1L contains: yeast powder 10g, peptone 20g, glucose 20g) at 30°C for 20h at 220r / min. Then connect in the 25L fermentation tank that 12L batch fermentation culture medium is housed by 10% inoculum size (1L contains: glucose 30g, ammonium sulfate 15g, magnesium sulfate 8g, potassium sulfate 7.5g, potassium dihydrogen phosphate 3.5g, calcium sulfate 1.0g, potassium hydroxide 1.2g, PTM 14mL), adjust the pH to 4.5 with ammonia water before inoculation. Maintain the dissolved oxygen concentration above 25% by controlling the air flow and speed. The batch culture ends after the carbon source is metabolized, and then the feed medium (1 L containing: 250 g of glucose) is added to make the bacteria continue to grow. During the 24 hours of fermentation. ...

Embodiment 2

[0024] The strain used was Pichia pastoris 602, provided by Wenzhou Conch Challenge Bioengineering Co., Ltd.

[0025] Pick a single colony from the plate and insert it into YPD seed medium (500mL Erlenmeyer flask liquid volume 125mL) (1L contains: yeast powder 10g, peptone 20g, glucose 20g) at 30°C for 20h at 220r / min. Then insert in the 25L fermentation tank that 12L batch fermentation culture medium is housed by 10% inoculum size (1L contains: glucose 50g, ammonium sulfate 20g, magnesium sulfate 9g, potassium sulfate 5g, potassium dihydrogenphosphate 2g, calcium sulfate 1.0g , potassium hydroxide 1.2g, PTM 14mL), adjust the pH to 4.7 with ammonia water before inoculation. Maintain the dissolved oxygen concentration above 25% by controlling the air flow and speed. The batch culture ends after the carbon source is metabolized, and the feed medium (1L containing: glucose 250g) is added to continue the growth of the bacteria, and the carbon source (glucose) is stopped after 24 ...

Embodiment 3

[0028] The strain used was Pichia pastoris 602, provided by Wenzhou Conch Challenge Bioengineering Co., Ltd.

[0029] Pick a single colony from the plate and insert it into YPD seed medium (500mL Erlenmeyer flask liquid volume 125mL) (1L contains: yeast powder 10g, peptone 20g, glucose 20g) at 30°C for 20h at 220r / min. Then connect in the 25L fermentation tank that 12L batch fermentation medium is housed by 10% inoculum size (1L contains: glucose 60g, ammonium sulfate 25g, magnesium sulfate 9g, potassium sulfate 9.5g, potassium dihydrogen phosphate 3.5g, calcium sulfate 0.8g, potassium hydroxide 1.2g, PTM 14mL), adjust the pH to 5.0 with ammonia water before inoculation. Maintain the dissolved oxygen concentration above 25% by controlling the air flow and speed. The batch culture ends after the carbon source is metabolized, and the feed medium (1L containing: glucose 250g) is added to continue the growth of the bacteria, and the carbon source (glucose) is stopped after 24 hou...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a process for preparing phytase by efficiently fermenting recombinant pichiapastoris and a special batch fermentation culture medium thereof. The process comprises a thalli growth stage and an exogenous gene induction expression stage and relates to promoters such as an alcohol oxidase promoter, a methanol promoter and the like. The culture medium in a fed-batch fermentation stage adopts a low-salt formula which comprises the following components in percentage by weight: 3 to 6 percent of carbon source, 1.5 to 3 percent of nitrogen source, 0.8 to 1.0 percent of magnesium sulfate, 0.5 to 1.0 percent of potassium sulfate and 0.3 to 0.4 percent of monopotassium phosphate, wherein fermentation temperature is between 27 and 30 DEG C; and fermentation pH is between 4.5 and 5.5. Methanol induction in the exogenous gene induction expression stage is performed in five stages. The preparation process of the invention has the advantages of lower cost and higher phytase fermentation activity.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, in particular to a process for preparing phytase by fermenting recombinant Pichia pastoris. Background technique [0002] Phytic acid widely exists in plant tissues and corresponding grain products, and 40%-70% of phosphorus contained in grain feed and its processed products is phytate phosphorus. All animal production is based on plant-based diets, and plant-based diets contain a large amount of phytic acid. The content of phytic acid in common feed ingredients is shown in Table 1. [0003] Table 1 Contents of phytic acid in common feed ingredients [0004] [0005] Note: The numbers in the table are based on dry matter and are taken from literature (W.Eeckhout and M.De Paepe.Total phosphorus, phytate phosphorus and phytase activity in plant feed stuffs.Anim Feed Sci Technol, 1994, 47:19-29.) [0006] Phytic acid (or phytate) in plant-based feeds is the primary storage form ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N9/16C12R1/84
Inventor 杨海龙吴明江林格郝名慧姜小伟
Owner WENZHOU UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com