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

Paper Details

Research Paper 01/04/2021
Views (411) Download (21)
current_issue_feature_image
publication_file

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

Ameen Ullah, Zulfiqar Ali Malik, Muhammad Irfan, Salah Ud Din, Qurratul Ain Rana, Malik Badshah1, Samiullah Khan, Fariha Hasan, Aamer Ali Shah
Int. J. Biosci.18( 4), 144-158, April 2021.
Certificate: IJB 2021 [Generate Certificate]

Abstract

Xylanases (EC 3.2.1.8) 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.

VIEWS 24

Amel BD, Nawel B, Khelifa B, Mohammed G, Manon J, Salima KG. 2016. Characterization of a purified thermostable xylanase from Caldicoprobacter algeriensis sp. nov. strain TH7C1T. Carbohydrate research 419, 60-8.

Archana A, Satyanarayana T. 2003. Purification and characterization of a cellulase-free xylanase of a moderate thermophile Bacillus licheniformis A99. World Journal of Microbiology and Biotechnology 19(1), 53-7.

Archana A, Sharma A, Satyanarayana T. 1999. Xylanolytic enzymes. Thermophilic Moulds in Biotechnology: Springer, p 169-90.

Bajaj BK, Manhas K. 2012. Production and characterization of xylanase from Bacillus licheniformis P11 (C) with potential for fruit juice and bakery industry. Biocatalysis and Agricultural Biotechnology 1(4), 330-7.

Basit A, Liu J, Miao T, Zheng F, Rahim K, Lou H. 2018. Characterization of Two Endo-β-1, 4-Xylanases from Myceliophthora thermophila and Their Saccharification Efficiencies, Synergistic with Commercial Cellulase. Frontiers in Microbiology 9, 233.

Chanwicha N, Katekaew S, Aimi T, Boonlue S. 2015. Purification and characterization of alkaline xylanase from Thermoascus aurantiacus var. levisporus KKU-PN-I2-1 cultivated by solid-state fermentation. mycoscience 56(3), 309-18.

Christov L, Myburgh J, O’Neill F, Van Tonder A, Prior B. 1999. Modification of the Carbohydrate Composition of Sulfite Pulp by Purified and Characterized β‐Xylanase and β‐Xylosidase of Aureobasidium pullulans. Biotechnology progress. 15(2), 196-200.

Collins T, Gerday C, Feller G. 2005. Xylanases, xylanase families and extremophilic xylanases. FEMS microbiology reviews 29(1), 3-23.

Damiano V, Bocchini D, Gomes E, Da Silva R. 2003. Application of crude xylanase from Bacillus licheniformis 77-2 to the bleaching of eucalyptus Kraft pulp. World Journal of Microbiology and Biotechnology 19(2), 139-44.

Demirjian DC, Morı́Varas F, Cassidy CS.  2001. Enzymes from extremophiles. Current opinion in chemical biology 5(2), 144-51.

Gessesse A, Gashe BA. 1997. Production of alkaline protease by an alkaliphilic bacteria isolated from an alkaline soda lake. Biotechnology letters. 19(5), 479.

Goswami GK, Rawat S. 2015. Microbial Xylanase and their applications-A review International journal current research and academic review 3(6), 436-50.

Haki G, Rakshit S. 2003. Developments in industrially important thermostable enzymes: a review. Bioresource technology 89(1), 17-34.

Irfan M, Guler HI, Belduz AO, Shah A, Canakci S. 2016. Cloning, purification and characterization of a cellulase-free xylanase from Geobacillus thermodenitrificans AK53. Applied biochemistry and microbiology 52(3), 277-86.

Irfan M, Guler HI, Ozer A, Sapmaz MT, Belduz AO, Hasan F. 2016. C-Terminal proline-rich sequence broadens the optimal temperature and pH ranges of recombinant xylanase from Geobacillus thermodenitrificans C5. Enzyme and microbial technology 91, 34-41.

Irfan M, Guler HI, Shah AA, Sal FA, Inan K, Belduz AO. 2016. Cloning, purification and characterization of halotolerant xylanase from Geobacillus thermodenitrificans C5. The Journal of Microbiology, Biotechnology and Food Sciences 5(6), 523.

Kallel F, Driss D, Bouaziz F, Neifer M, Ghorbel R, Chaabouni SE. 2015. Production of xylooligosaccharides from garlic straw xylan by purified xylanase from Bacillus mojavensis UEB-FK and their in vitro evaluation as prebiotics. Food and Bioproducts Processing 94, 536-46.

Kocabaş DS, Güder S, Özben N. 2015. Purification strategies and properties of a low-molecular weight xylanase and its application in agricultural waste biomass hydrolysis. Journal of Molecular Catalysis B: Enzymatic 115, 66-75.

Kumar V, Satyanarayana T. 2014. Production of thermo-alkali-stable xylanase by a novel polyextremophilic Bacillushalodurans TSEV1 in cane molasses medium and its applicability in making whole wheat bread. Bioprocess and biosystems engineering 37(6), 1043-53.

Laemmli V. 1970. Determination of protein molecular weight in polyacrylamide gels. Nature. 227, 680-5.

Lineweaver H, Burk D. 1934. The determination of enzyme dissociation constants. Journal of the American chemical society 56(3), 658-66.

Lombard V, Golaconda Ramulu H, Drula E, Coutinho PM, Henrissat B. 2013. The carbohydrate-active enzymes database (CAZy) in 2013. Nucleic acids research 42(D1), D490-D5.

Marcolongo L, La Cara F, Morana A, Di Salle A, Del Monaco G, Paixão SM. 2015. Properties of an alkali-thermo stable xylanase from Geobacillus thermodenitrificans A333 and applicability in xylooligosaccharides generation. World Journal of Microbiology and Biotechnology 31(4), 633-48.

Nihalani D, Satyanarayana T. 1992. Isolation and characterization of extracellular alkaline enzyme producing bacteria. Indian journal of microbiology New Delhi 32(4), 443-9.

Ninawe S, Kapoor M, Kuhad RC. 2008. Purification and characterization of extracellular xylanase from Streptomyces cyaneus SN32. Bioresource Technology 99(5), 1252-8.

Saitou N, Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular biology and evolution. 4(4), 406-25.

Techapun C, Charoenrat T, Watanabe M, Sasaki K, Poosaran N. 2002. Optimization of thermostable and alkaline-tolerant cellulase-free xylanase production from agricultural waste by thermotolerant Streptomyces sp. Ab106, using the central composite experimental design. Biochemical Engineering Journal 12(2), 99-105.

Touzel JP, O’Donohue M, Debeire P, Samain E, Breton C. 2000. Thermobacillus xylanilyticus gen. nov., sp. nov., a new aerobic thermophilic xylan-degrading bacterium isolated from farm soil. International journal of systematic and evolutionary microbiology 50(1), 315-20.

van Dyk JS, Sakka M, Sakka K, Pletschke BI. 2009. The cellulolytic and hemi-cellulolytic system of Bacillus licheniformis SVD1 and the evidence for production of a large multi-enzyme complex. Enzyme and Microbial Technology 45(5), 372-8.

Vieille C, Zeikus GJ. 2001. Hyperthermophilic enzymes: sources, uses, and molecular mechanisms for thermostability. Microbiology and molecular biology reviews 65(1), 1-43.

Zheng HC, Sun MZ, Meng LC, Pei HS, Zhang XQ, Yan Z. 2014. Purification and characterization of a thermostable xylanase from Paenibacillus sp. NF1 and its application in xylooligosaccharides production. J Microbiol Biotechnol  24(4), 489-96.

Zhu Y, Li X, Sun B, Song H, Li E, Song H. 2012. Properties of an Alkaline‐Tolerant, Thermostable Xylanase from Streptomyces chartreusis L1105, Suitable for Xylooligosaccharide Production. Journal of food science 77(5).