A metalloprotease molecular cloning and sequencing of Anoxybacillus flavithermus from Qinarjeh, Meshgin-Shahr, Iran

Document Type : Research Paper

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Abstract

Biomacromolecules of thermophilic organisms are often thermostable. Apart from high temperature they are also known to withstand denaturants of extremely acidic and alkaline conditions. Thermostable enzymes have capable potentioal in the food and paper industries, detergents, drugs, toxic wastes removal and oil drilling. The enzymes can be produced from the thermophiles through either optimized fermentation of the microorganisms or cloning of fast-growing mesophiles by recombinant DNA technology. Metalloproteases are metal ion dependent protease. Metalloproteases that used in this study was a zinc-dependent endopeptidase. Metalloprotease gene was cloned from Anoxybacillus flavithermus. For the amplification of the gene, PCR was performed. A pair of primers designed with a cleavage site for enzymes XhoI and NheI and vector pET21a(+) clone was transformed into BL21 host. The results of the cloned gene sequence analysis showed similarity to the protease gene different variants of the species Anoxybacillus flavithermus.

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