Optimization of production and partial characterization of xylanase from a newly isolated Bacillus amyloliquefaciens

Bruno Las-Casas Chaves, Ana Paula Martinazzo, Brisabella Coca, Adriane Nunes de Souza, Carlos Eduardo Teodoro

Abstract


This paper reports the process of production optimization and partial characterization of xylanase from a newly isolated Bacillus amyloliquefacies VR002, isolated from local soil. The microorganism exhibited maximum xylanase production when 1.0% (v/v) of inoculum size was added to culture medium with initial pH 6, 1.0% (w/v) birchwood xylan, at 35 °C after 48h of incubation. Xylanase production in different carbon sources apart from birchwood xylan and xylose did not show high production levels. Optimum pH for xylanase activity was 6.0. The enzyme was alkali-stable and retained 100% of residual activity over the pH range from 6.0 to 10.0 for 24 h at 25°C. Optimum temperature for enzyme activity was 55°C. Xylanase was 100% stable at 4°C and 25°C even after 24h of incubation, a desirable characteristic for enzyme storage. Moreover, best crude extract volume and time reaction were found to be 10 µL and 5 min, respectively. After optimization of production and activity parameters, an increase of nearly 60-fold in xylanase activity (44.12 ± 4.36 U/mL) was achieved. Characteristics of B. amyloliquefaciens VR002 xylanase are particularly desirable for biotechnological applications

Keywords


xylanase; bacillus amyloliquefaciens; optimization

Full Text:

HTML

References


ADHYARU, Dharmesh N. et al. Cellulase-free-thermo-alkali-solvent-stable xylanase from Bacillus altitudinis DHN8: Over-production through statistical approach, purification and bio-deinking/ bio-bleaching potential. Biocatalysis and Agricultural Biotechnology, [S. l.], v. 12, p. 220–227, 2017. Disponível em: https://doi.org/10.1016/j.bcab.2017.10.010

AKHAVAN SEPAHY, Abbas; GHAZI, Shokoofeh; AKHAVAN SEPAHY, Maryam. Cost-effective production and optimization of alkaline xylanase by indigenous Bacillus mojavensis AG137 fermented on agricultural waste. Enzyme Research, [S. l.], v. 2011, n. 1, 2011.

ALTAF, Simair A. et al. Characterization of Crude Xylanase Produced by Edible Mushroom Pleurotus eryngii. Journal of bioprocessing and biotechniques, [S. l.], v. 6, n. 2, p. 6–11, 2016.

BAJAJ, Bijender Kumar; MANHAS, Kalpana. Production and characterization of xylanase from Bacillus licheniformis P11(C) with potential for fruit juice and bakery industry. Biocatalysis and Agricultural Biotechnology, [S. l.], v. 1, n. 4, p. 330–337, 2012. Disponível em: http://dx.doi.org/10.1016/j.bcab.2012.07.003

BATTAN, Bindu et al. Enhanced production of cellulase-free thermostable xylanase by Bacillus pumilus ASH and its potential application in paper industry. Enzyme and Microbial Technology, [S. l.], v. 41, n. 6–7, p. 733–739, 2007.

BÉRENGER, Jean-François et al. Production, purification, and properties of thermostable xylanase from Clostridium stercorarium . Canadian Journal of Microbiology, [S. l.], v. 31, n. 7, p. 635–643, 2010.

BERNIER, Roger et al. Isolation and Characterization of a Xylanase from Bacillus subtilis. Applied and environmental microbiology, [S. l.], v. 46, n. 2, p. 511–514, 1983. Disponível em: http://aem.asm.org/content/46/2/511.short

BHALLA, Aditya; BISCHOFF, Kenneth M.; SANI, Rajesh Kumar. Highly Thermostable Xylanase Production from A Thermophilic Geobacillus sp. Strain WSUCF1 Utilizing Lignocellulosic Biomass. Frontiers in Bioengineering and Biotechnology, [S. l.], v. 3, n. June, p. 1–8, 2015.

BOCCHINI, D. .. et al. Optimization of Xylanase Production by Bacillus Circulans D1 in Submerged Fermentation Using Response Surface Methodology. Process Biochemistry, [S. l.], v. 38, n. 5, p. 727–731, 2003.

BOCCHINI, D. A.; GOMES, E.; DA SILVA, R. Xylanase production by Bacillus circulans D1 using maltose as carbon source. Applied Biochemistry and Biotechnology, [S. l.], v. 146, n. 1–3, p. 29–37, 2008.

COUGHLAN, MP; HAZLEWOOD, GP. beta‐1,4‐D‐xylan‐degrading enzyme systems: biochemistry, molecular biology and applications. Biotechnology and Applied Biochemistry, [S. l.], v. 17, n. 3, p. 259–289, 1993.

DEEKER, Robert F. H.; ROBERTS, Geoffrey N. Structures of the oligosaccharides from the enzymic hydrolysis of hemicellulose by a hemicellulase of Ceratocystis paradoxa. Carbohydrate Research, [S. l.], v. 43, p. 335–344, 1975.

DHILLON, Ashita et al. A cellulase-poor, thermostable, alkalitolerant xylanase produced by Bacillus circulans AB 16 grown on rice straw and its application in biobleaching of eucalyptus pulp. Bioresource Technology, [S. l.], v. 73, n. 3, p. 273–277, 2000.

GHOSH, Mousumi et al. Aspergillus sydowii MG 49 is a strong producer of thermostable xylanolytic enzymes. Enzyme and Microbial Technology, [S. l.], v. 15, n. 8, p. 703–709, 1993.

GUPTA, Uma; KAR, Rita. Xylanase production by a thermo-tolerant Bacillus species under solid-state and submerged fermentation. Brazilian Archives of Biology and Technology, [S. l.], v. 52, n. 6, p. 1363–1371, 2009.

HALTRICH, Dietmar et al. Production of fungal xylanases. Bioresource Technology, [S. l.], v. 58, n. 2, p. 137–161, 1996.

HECK, Júlio Xandro; DE BARROS SOARES, Luís Henrique; ZÁCHIA AYUB, Marco Antônio. Optimization of xylanase and mannanase production by Bacillus circulans strain BL53 on solid-state cultivation. Enzyme and Microbial Technology, [S. l.], v. 37, n. 4, p. 417–423, 2005.

IRFAN, Muhammad et al. Optimization of process parameters for xylanase production by Bacillus sp. in submerged fermentation. Journal of Radiation Research and Applied Sciences, [S. l.], v. 9, n. 2, p. 139–147, 2015. Disponível em: http://dx.doi.org/10.1016/j.jrras.2015.10.008

KAPOOR, Mukesh; NAIR, Lavanya M.; KUHAD, Ramesh Chander. Cost-effective xylanase production from free and immobilized Bacillus pumilus strain MK001 and its application in saccharification of Prosopis juliflora. Biochemical Engineering Journal, [S. l.], v. 38, n. 1, p. 88–97, 2008.

KAVYA, Venkatesh; PADMAVATHI, Tallapragada. Optimization of growth conditions for xylanase production by Aspergillus niger in solid state fermentation. Polish Journal of Microbiology, [S. l.], v. 58, n. 2, p. 125–130, 2009.

KHASIN, Alexander; ALCHANATI, Iris; SHOHAM, Yuval. Purification and Characterization of a Thermostable Xylanase from Bacillus stearothermophilus T-6. Applied and Environmental Microbiology, [S. l.], v. 59, n. 6, p. 1725–1730, 1993.

KHUSRO, Ameer et al. Statistical optimization of thermo-alkali stable xylanase production from Bacillus tequilensis strain ARMATI. Electronic Journal of Biotechnology, [S. l.], v. 22, p. 16–25, 2016. Disponível em: http://dx.doi.org/10.1016/j.ejbt.2016.04.002

KULKARNI, Neeta; SHENDYE, Abhay; RAO, Mala. Molecular and biotechnological aspects of xylanases. FEMS Microbiology Reviews, [S. l.], v. 23, n. 4, p. 411–456, 1999.

KUMAR, Sharad et al. Purification, characterization and thermostability improvement of xylanase from Bacillus amyloliquefaciens and its application in pre-bleaching of kraft pulp. 3 Biotech, [S. l.], v. 7, n. 1, 2017.

LAMA, Licia et al. Purification and characterization of thermostable xylanase and β-xylosidase by the thermophilic bacterium Bacillus thermantarcticus. Research in Microbiology, [S. l.], v. 155, n. 4, p. 283–289, 2004.

LENGELER, J. W. Catabolite Repression. Brenner’s Encyclopedia of Genetics: Second Edition, [S. l.], p. 447–452, 2013.

LINCOLN, Ralph E. Control of stock culture preservation and inoculum build-up in bacterial fermentation. Journal of Biochemical and Microbiological Technology and Engineering, [S. l.], v. 2, n. 4, p. 481–500, 1960.

MILLER, GL. Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Analytical Chemistry, [S. l.], v. 31, n. 3, p. 426–428, 1959.

MOON, Seung‐Hyeon ‐H; PARULEKAR, Satish J. A parametric study ot protease production in batch and fed‐batch cultures of Bacillus firmus. Biotechnology and Bioengineering, [S. l.], v. 37, n. 5, p. 467–483, 1991.

MURUGAN, Sevanan et al. Production of Xylanase from Arthrobacter sp. MTCC 6915 Using Saw Dust As Substrate under Solid State Fermentation . Enzyme Research, [S. l.], v. 2011, p. 1–7, 2011.

NAGAR, Sushil et al. Production and optimization of cellulase-free, alkali-stable xylanase by Bacillus pumilus SV-85S in submerged fermentation. Journal of Industrial Microbiology and Biotechnology, [S. l.], v. 37, n. 1, p. 71–83, 2010.

NAGAR, Sushil et al. Production of alkali tolerant cellulase free xylanase in high levels by Bacillus pumilus SV-205. International Journal of Biological Macromolecules, [S. l.], v. 50, n. 2, p. 414–420, 2012. Disponível em: http://dx.doi.org/10.1016/j.ijbiomac.2011.12.026

NASCIMENTO, R. P. et al. Production and partial characterisation of xylanase from Streptomyces sp. strain AMT-3 isolated from Brazilian cerrado soil. Enzyme and Microbial Technology, [S. l.], v. 31, n. 4, p. 549–555, 2002.

POLIZELI, M. L. T. M. et al. Xylanases from fungi: Properties and industrial applications. Applied Microbiology and Biotechnology, [S. l.], v. 67, n. 5, p. 577–591, 2005.

PRAKASH, P. et al. Production of alkaliphilic, halotolerent, thermostable cellulase free xylanase by Bacillus halodurans PPKS-2 using agro waste: Single step purification and characterization. World Journal of Microbiology and Biotechnology, [S. l.], v. 28, n. 1, p. 183–192, 2012.

RANI, D. Swaroopa; NAND, Krishna. Production of thermostable cellulase-free xylanase by Clostridium absonum CFR-702. Process Biochemistry, [S. l.], v. 36, n. 4, p. 355–362, 2000.

RATANAKHANOKCHAI, Khanok; KYU, Khin Lay; TANTICHAROEN, Morakot. Purification and Properties of a Xylan-Binding Endoxylanase from Alkaliphilic Bacillus sp . Strain K-1. Applied and environmental microbiology, [S. l.], v. 65, n. 2, p. 694–697, 1999.

REYMOND, Jean Louis; FLUXÀ, Viviana S.; MAILLARD, Noélie. Enzyme assays. Chemical Communications, [S. l.], v. 1, n. 1, p. 34–46, 2009. Disponível em: http://dx.doi.org/10.1016/j.pisc.2014.02.005

SANGHI, Ashwani et al. One-step purification and characterization of cellulase-free xylanase produced by alkalophilic bacillus subtilis ASH. Brazilian Journal of Microbiology, [S. l.], v. 41, n. 2, p. 467–476, 2010.

SCANDURRA, Roberto et al. Protein thernmstability in extrernophiles. Biochimie, [S. l.], v. 80, n. D, p. 933–941, 1998.

SHAH, Amita R.; MADAMWAR, Datta. Xylanase production by a newly isolated Aspergillus foetidus strain and its characterization. Process Biochemistry, [S. l.], v. 40, n. 5, p. 1763–1771, 2005.

SRINIVASAN, M. C.; RELE, Meenakshi V. Microbial xylanase for paper industry. Current Science, [S. l.], v. 77, n. 1, p. 137–142, 1999.

SUNNA, A.; ANTRANIKIAN, G. Xylanolytic Enzymes from Fungi and Bacteria. Critical Reviews in Biotechnology, [S. l.], v. 17, n. 1, p. 39–67, 2008.




DOI: http://dx.doi.org/10.5902/2179460X42741

Refbacks

  • There are currently no refbacks.


Copyright (c) 2020 Ciência e Natura

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.