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Original Article
- Cloning, Expression, and Purification of Hyperthermophile α-Amylase from
Pyrococcus woesei - Amir Ghasemi, Sobhan Ghafourian, Sedighe Vafaei, Reza Mohebi, Maryam Farzi, Morovat Taherikalani, Nourkhoda Sadeghifard
- Osong Public Health Res Perspect. 2015;6(6):336-340. Published online December 31, 2015
- DOI: https://doi.org/10.1016/j.phrp.2015.10.003
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- 3 Crossref
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Abstract
PDF - 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. -
Citations
Citations to this article as recorded by
- Escherichia coli expression and characterization of α-amylase from Geobacillus thermodenitrificans DSM-465
A. Al-Amri, M. A. Al-Ghamdi, J. A. Khan, H. N. Altayeb, H. Alsulami, M. Sajjad, O. A. Baothman, M. S. Nadeem
Brazilian Journal of Biology.2022;[Epub] CrossRef - Glycoside Hydrolases and Glycosyltransferases from Hyperthermophilic Archaea: Insights on Their Characteristics and Applications in Biotechnology
Khadija Amin, Sylvain Tranchimand, Thierry Benvegnu, Ziad Abdel-Razzak, Hala Chamieh
Biomolecules.2021; 11(11): 1557. CrossRef - Optimization, Purification, and Starch Stain Wash Application of Two Newα-Amylases Extracted from Leaves and Stems ofPergularia tomentosa
Imen Lahmar, Hanen El Abed, Bassem Khemakhem, Hafedh Belghith, Ferjani Ben Abdallah, Karima Belghith
BioMed Research International.2017; 2017: 1. CrossRef
- Escherichia coli expression and characterization of α-amylase from Geobacillus thermodenitrificans DSM-465
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