Heat stability improvement and efficient expression of phytase with high specific activity

A high-efficiency expression and thermal stability technology, applied in the field of improved phytase and its encoding gene, can solve the problems of poor thermal stability and large loss

Inactive Publication Date: 2005-12-14
安徽固源生物工程有限责任公司
View PDF0 Cites 44 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The optimal temperature of APPA is 60°C, but its thermal stability is poor. The remaining enzyme activity is o

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
  • Heat stability improvement and efficient expression of phytase with high specific activity
  • Heat stability improvement and efficient expression of phytase with high specific activity
  • Heat stability improvement and efficient expression of phytase with high specific activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0024] Experimental conditions

[0025] 1. Strains and vectors Escherichia coli strain E.coli DH5a and plasmid pET-22b(+) were purchased from Promega, and yeast strain Pichia pastoris GS115(His-Mut+) and plasmid pFF01 were kindly donated by Dr. D.Luo, University of Alberta, Canada.

[0026] 2. Enzymes and kits Restriction enzymes, ligases, Taq enzymes, and DNAaseI are products of Boehringer. PCR Kits were purchased from Promega.

[0027] 3. Biochemical reagents DNA synthesis reagents are products of Milipore Company. Primers were synthesized with ABI Cyclone DNA synthesizer. IPTG, X-Gal, SDS and sodium phytate are products of Sigma. TEMED, ammonium persulfate, acrylamide and methylene bisacrylamide are products of Promega.

[0028]4. Medium The culture medium for Escherichia coli is LB (1% peptone, 0.5% yeast extract, 1% NaCl, pH 7.0). Yeast complete medium is YPD (1% yeast extract, 2% peptone, 2% glucose); yeast transformation medium is RDB [18.6% sorbitol, 2% glucose, 1...

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 present invention provides one kind of improved high specific activity phytase and its coding gene, and the improved phytase has even higher heat stability. The present invention also provides recombinant yeast cell containing the phytase gene of the present invention. The recombinant yeast of the present invention has phytase expressing amount up to 6.5E6 U in each ml of fermented liquid. The phytase of the present invention may be used widely in feed and food industry.

Description

technical field [0001] The invention relates to an improved phytase and its coding gene. The invention also relates to a recombinant yeast cell containing the phytase gene and capable of highly expressing the phytase. Background technique [0002] Phytase (EC.3.1.3.8) is an enzyme that can hydrolyze phytic acid. It degrades phytate phosphorus into inositol and phosphoric acid. Phytase widely exists in microorganisms, such as Aspergillus in fungi (Yamada K. et al. Agric. Biol. Chem. 32: 1275-1282, 1986; Van Gorcom R.F.M. et al., US Patent No. 5436156, 1995) , Yeast (Nayini N.R.et al.Lebensm Wiss.Technol.17:24-26,1984); Bacillus subtilis (Paver, V.K.J., Bacteriol.151:1102-1108,1982) in bacteria, Pseudomonas (Cosgrove D.J., Austral.J.Biol.Sci.23:1207-1220, 1970), Lactobacillus (Shirai K., Letters Appl.Biol.Sci.19:366-369, 1994), Escherichia coli (Greiner R., Arch Biochem. Biophys. 303:107-113, 1993) et al. [0003] Phytase can be widely used in monogastric animal feed (War...

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): C12N9/16
Inventor 孙跃军孙齐军
Owner 安徽固源生物工程有限责任公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap