(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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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 | ||||||||||||||||
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| Keywords: | amylase expression recombinant protein thermophile | ||||||||||||||||
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| 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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