Repository of Research and Investigative Information

Repository of Research and Investigative Information

Ilam University of Medical Sciences

Cloning, Expression, and Purification of Hyperthermophile α-Amylase from Pyrococcus woesei

Fri May 20 02:01:24 2022

(2015) Cloning, Expression, and Purification of Hyperthermophile α-Amylase from Pyrococcus woesei. Osong public health and research perspectives. pp. 336-340. ISSN 22109099 (ISSN)

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Official URL: https://www.scopus.com/inward/record.uri?eid=2-s2....

Abstract

Objectives: In an attempt α-amylase gene from Pyrococcus woesei was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY- α-amylase. Methods: Escherichia coli BL21 used as a host and protein expression was applied using IPTG. SDS-PAGE assay demonstrated the 100 kDa protein. Amylolytic activity of proteins produced by transformed E. coli cells was detected by zymography, and the rate of active α-amylase with and without the intein tag in both soluble conditions and as inclusion bodies solubilized by 4M urea were measured. Results: Amylolytic activity of ∼185,000 U/L of bacterial culture was observed from the soluble form of the protein using this system. Conclusion: These results indicate that this expression system was appropriate for the production of thermostable α-amylase. © 2015.

Item Type: Article
Creators:
CreatorsEmail
Ghasemi, A.UNSPECIFIED
Ghafourian, S.UNSPECIFIED
Vafaei, S.UNSPECIFIED
Mohebi, R.UNSPECIFIED
Farzi, M.UNSPECIFIED
Taherikalani, M.UNSPECIFIED
Sadeghifard, N.UNSPECIFIED
Keywords: Amylase Expression Recombinant protein Thermophile recombinant enzyme alpha amylase gene Article bacterial gene bacterium culture cell inclusion controlled study enzyme activity enzyme purification Escherichia coli molecular cloning molecular weight nonhuman polyacrylamide gel electrophoresis priority journal protein expression Pyrococcus furiosus recombinant plasmid solubility zymography
Divisions:
Page Range: pp. 336-340
Journal or Publication Title: Osong public health and research perspectives
Journal Index: Scopus
Volume: 6
Number: 6
Identification Number: https://doi.org/10.1016/j.phrp.2015.10.003
ISSN: 22109099 (ISSN)
Depositing User: مهندس مهدی شریفی
URI: http://eprints.medilam.ac.ir/id/eprint/1437

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