Preparing purification method for thermophilic alpha-amylase

A technology of amylase and thermophilic archaea, applied in the field of bioengineering

Inactive Publication Date: 2007-10-31
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the factors of yield, the glycerol extraction method also has problems in process amplification and other aspects

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
  • Preparing purification method for thermophilic alpha-amylase
  • Preparing purification method for thermophilic alpha-amylase
  • Preparing purification method for thermophilic alpha-amylase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Preparation of thermophilic alpha-amylase PFA

[0063] Materials and Methods

[0064] 1. Materials

[0065] Strains and vectors: Genomic DNA of Pyrococcus furiosus DSM3638 ATCC No. 43587 TM ; Escherichia coli E.coli DH5α, E.coli BL21(DE3) and E.coli BL21-CodonPlus(DE3)-RIL were purchased from Stratagen; pET21a expression vector was purchased from Novagen;

[0066] Molecular biology reagents: Restriction enzymes Nde I, HindIII, Pyrobest DNA polymerase, T4 DNA polymerase, T4 DNA ligase were purchased from TaKaRa Company; protein molecular quality standards were purchased from Shanghai Institute of Biochemistry; Phenyl Sepharose 6 Fast Flow chromatography The column material was purchased from Novagen; the gel recovery kit was purchased from Huashun Bioengineering Co., Ltd.; the primer synthesis was completed by Shanghai Bocai Biotechnology Co., Ltd.; the DNA sequencing was completed by Yingjun Biotechnology Co., Ltd.; other reagents were of analytical grade.

[0067] 2...

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 preparing method of thermophilic alpha-amylase (PFA), which comprises the following steps: heating and dissolving recombinant protein inclusion body under the condition of alkalinity; diluting; compositing; separating and purifying through column layer. This method possesses high reclaiming ratio and high alpha-amylase activity.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the extraction and purification of recombinant proteins and thermophilic enzymes. Background technique [0002] High temperature resistant α-amylase is one of the most commonly used enzyme preparations in industry, and is widely used in starch processing in food industry and chemical industry. The search and development of high heat-tolerant amylases has long been a key and hot topic in the field of industrial enzyme research. The global starch processing enzyme market is about 150 million US dollars, of which about 24% are used in the starch liquefaction process. Therefore, any improvement in amylase production yield, heat resistance and enzyme activity has very important theoretical significance and direct commercial value. [0003] At present, the heat-resistant amylase commonly used in industry comes from Bacillus licheniformis. The optimum catalytic temperature of t...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/28C12P19/14C12P19/02
Inventor 张毅王丽萨杨胜利
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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