Repository of Research and Investigative Information

Repository of Research and Investigative Information

Ilam University of Medical Sciences

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

Mon Oct 3 02:33:49 2022

(2015) Cloning, Expression, and Purification of Hyperthermophile alpha-Amylase from Pyrococcus woesei. Osong public health and research perspectives. pp. 336-40. ISSN 2210-9099 (Print) 2210-9099 (Linking)

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Official URL: https://www.ncbi.nlm.nih.gov/pubmed/26835242

Abstract

OBJECTIVES: In an attempt alpha-amylase gene from Pyrococcus woesei was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY- alpha-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 alpha-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 approximately 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 alpha-amylase.

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
Divisions:
Page Range: pp. 336-40
Journal or Publication Title: Osong public health and research perspectives
Journal Index: Pubmed
Volume: 6
Number: 6
Identification Number: https://doi.org/10.1016/j.phrp.2015.10.003
ISSN: 2210-9099 (Print) 2210-9099 (Linking)
Depositing User: مهندس مهدی شریفی
URI: http://eprints.medilam.ac.ir/id/eprint/1080

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