Cloning and sequencing xylanase ofAnoxybacillus flavithermus from Qinarjeh, Meshkin-Shahr, Iran

The thermostable and alkaliphile xylanases are beneficial industrial enzymes in food industry (bakery and alcohol production or fermentation industries) pulp-kraft, and pharmaceutic industry. Xylanases hydrolyse xylan (the hemicellulose of plant cell wall) and produced by different kinds of microorganisms like bacteria, fungi and some algae. In this study, xylanase gene from thermoalkaline Anoxybacillus flavithermus was isolated from hot spring of Meshkinshahr (Iran) and was cloned in E. coli (BL21). At first, total DNA was extracted from bacteria. PCR was then accomplished and xylanase gene was isolated and amplified. Amplification of xylenase coding gene was confirmed by electrophoresis. Cloning was performed by digestion of xylanase gene and pET21a by restriction enzymes (NheI and XhoI), following by ligation and transformation to E. coli (BL21). Finally, the results were confirmed by colony-PCR and sequencing of selected colonies. PCR and sequencing results, demonstrated a high similarity between xylanases of Anoxybacillus species with xylanase of this study.