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Production and characterization of xylanase from Bacillus licheniformis S3 isolated from hot spring

Research Paper | April 1, 2021

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Ameen Ullah, Zulfiqar Ali Malik, Muhammad Irfan, Salah Ud Din, Qurratul Ain Rana, Malik Badshah1, Samiullah Khan, Fariha Hasan, Aamer Ali Shah

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Int. J. Biosci.18( 4), 144-158, April 2021

DOI: http://dx.doi.org/10.12692/ijb/18.4.144-158


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Xylanases (EC are hemicellulases responsible for the catalysis of xylan, a major component of hemicellulose and the second largest renewable biomass. It is applied in various industries such as paper and pulp, biofuel, baking, detergents, animal feeds and textile. The current study was focused on screening of hot spring samples, collected from Skardu, Pakistan, for novel xylanases producing microbial strains. A bacterium designated as strain S3 was isolated that could effectively breakdown xylan. It was found to be the specie of genus Bacillus based on morphological, biochemical analysis, while 16S rRNA gene sequencing results indicated 99% similarity with Bacillus licheniformis. Various physical and chemical conditions were statistically optimized using Design-expert software for maximum production of xylanase. Xylanase produced under these optimized physical and chemical conditions, was purified to homogeneity by size exclusion column chromatography using Sephadex G-100. The molecular weight was found to be approximately 28 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The specific activity of the purified xylanase was up to 39 U/mg with a 4.38-fold purification and 58% yield. The Km and Vmax of S3 xylanase were 8.6 mgmL−1 and 43.71 μmolmg−1min−1, respectively. The activity exhibited by B. licheniformis xylanase was found to be acellulolytic with stability at wide temperature (40°C-60°C) and pH (5.0 to 10.0), and stimulated by CaCl2, FeSO4, FeSO4, CdCl2 and MgSO4, while inhibited by HgCl2 and CuSO4. Furthermore, the enzyme was resistant to most of the proteases tested. Since the enzyme was stable at wide pH range (5.0-10.0) and showed resistance to Ni, Cd, Zn and Co at 10 mM concentration that represents its efficiency and potential application in paper and pulp, food and feed industries.


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Production and characterization of xylanase from Bacillus licheniformis S3 isolated from hot spring

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