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Characterization, cloning and functional expression of novel xylanase from Thermomyces lanuginosus SS-8 isolated from self-heating plant wreckage material

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Abstract

Extracellular cellulase free xylanase from Thermomyces lanuginosus sp. SS-8, isolated from self heating plant wreckage material was identified as β-1,4-endo-xylanase precursor, a monomer of 21.3 kDa with no carbohydrate residue. This xylanase retained 80 % activity at 60 °C for 96 h, was active at a wide pH range of 3–11 and uniquely hydrolyzed xylan to xylose without production of xylo-oligosaccharides. Gene xynSS8 encoding xylanase from T. lanuginosus SS-8 was cloned and functionally expressed in Escherichia coli XL1 Blue using pTZ57R/T plasmid and xynSS8/pQE-9 expression vector construct respectively. Gene xynSS8 was of 777 bp and deduced amino acid sequence was a mature xylanase of 258 amino acids. XynSS8 has extra 33 amino acids compared to its nearest homolog and was thermo-alkali tolerant as that of native protein. The xylanase could degrade pulp and release substantial chromophoric materials and lignin derived compounds indicating its effective utility in pulp bleaching. Novel characteristics of the enzyme may contribute to its wide industrial usage. This is first report of cloning and functional expression of the novel xylanase from T. lanuginosus SS-8.

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References

  • Beg QK, Kapoor M, Mahajan L, Hoondal GS (2001) Microbial xylanases and their industrial applications: a review. Appl Microbiol Biotechnol 56:326–338

    Article  CAS  Google Scholar 

  • Biely P (1985) Microbial xylanolytic systems. Trends Biotechnol 3:286–290

    Article  CAS  Google Scholar 

  • Bissoon S, Christov L, Singh S (2002) Bleach boosting effects of purified xylanase from Thermomyces lanuginosus SSBP on bagasse pulp. Process Biochem 37:567–572

    Article  CAS  Google Scholar 

  • Blum H, Beier H, Gross B (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Deng P, Li D, Cao Y, Lu W, Wang C (2006) Cloning of a gene encoding an acidophilic endo-β-1,4-xylanase obtained from Aspergillus niger CGMCC1067 and constitutive expression in Pichia pastoris. Enzyme Microb Technol 39:1096–1102

    Article  CAS  Google Scholar 

  • Do TT, Quyen DT, Dam TH (2012) Purification and characterization of an acid-stable and organic solvent-tolerant xylanase from Aspergillus awamori VTCC-F312. ScienceAsia 38:157–165

    Article  CAS  Google Scholar 

  • Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 58:350–356

    Article  Google Scholar 

  • Gaffney M, Carberry S, Doyle S, Murphy R (2009) Purification and characterisation of a xylanase from Thermomyces lanuginosus and its functional expression by Pichia pastoris. Enzyme Microb Technol 45:348–354

    Article  CAS  Google Scholar 

  • Hudson HJ (1992) Fungal biology. Cambridge University Press, Cambridge, pp 106–170

    Google Scholar 

  • Hung KS, Liu SM, Tzou WS, Lin FP, Pan CL, Fang TY, Sun KH, Tang SJ (2011) Characterization of a novel GH10 thermostable, halophilic xylanase from the marine bacterium Thermoanaerobacterium saccharolyticum NTOU1. Process Biochem 46:1257–1263

    Article  CAS  Google Scholar 

  • Ichinose H, Diertavitian S, Fujimoto Z, Kuno A, Leggio LL, Kaneko S (2011) Structure-based engineering of glucose specificity in a family 10 xylanase from Streptomyces olivaceoviridis E-86. Process Biochem 47:358–365

    Article  Google Scholar 

  • Laemlli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  Google Scholar 

  • Li L, Tian H, Cheng Y, Jiang Z, Yang S (2006) Purification and characterization of a thermostable cellulase-free xylanase from the newly isolated Paecilomyces themophila. Enzyme Microb Technol 38:780–787

    Article  CAS  Google Scholar 

  • Lineweaver H, Burk D (1934) The determination of the enzyme dissociation. J Am Chem Soc 56:658–666

    Article  CAS  Google Scholar 

  • Miller GH (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–429

    Article  CAS  Google Scholar 

  • Ornstein L, Davis BJ (1964) Disc electrophoresis. I. Background and theory. Ann NY Acad Sci 121:321–349

    Article  CAS  Google Scholar 

  • Prade RA (1995) Xylanases: from biology to biotechnology. Biotechnol Genet Eng Rev 13:101–131

    Article  Google Scholar 

  • Royer JC, Nakas JP (1990) Simple, sensitive zymogram technique for detection of xylanase activity in polyacrylamide gels. Appl Environ Microbiol 56:1516–1517

    CAS  Google Scholar 

  • Ruanglek V, Sriprang R, Ratanaphan N, Tirawongsaroj P, Chantasigh D, Tanapongpipat S, Pootanakit K, Eurwilaichitr L (2007) Cloning, expression, characterization, and high cell-density production of recombinant endo-1,4-β-xylanase from Aspergillus niger in Pichia pastoris. Enzyme Microb Technol 41:19–25

    Article  CAS  Google Scholar 

  • Ryan SE, Nolan K, Thompson R, Gubitz GM, Savage AV, Tuohy MG (2003) Purification and characterization of a new low molecular weight endoxylanase from Penicillium capsulatum. Enzyme Microb Technol 33:775–785

    Article  CAS  Google Scholar 

  • Schlacher A, Holzmann K, Hayn M, Steiner W, Schwab H (1996) Cloning and characterization of the gene for the thermostable xylanase XynA from Thermomyces lanuginosus. J Biotechnol 49:211–218

    Article  CAS  Google Scholar 

  • Sharma M, Kumar A (2013) Xylanases: an overview. Br Biotechnol J 3:1–28

    CAS  Google Scholar 

  • Shrivastava S, Shukla P, Mukhopadhayay K (2008) Correlative characterization of changes in hyphal morphology during xylanase production in submerged culture by Thermomyces lanuginosus SS-8. Internet J Microbiol 4:2

    Google Scholar 

  • Shrivastava S, Poddar R, Shukla P, Mukhopadhyay K (2009) Study of codon bias perspective of fungal xylanase gene by multivariate analysis. Bioinformation 3:425–429

    Article  Google Scholar 

  • Shrivastava S, Shukla P, Mukhopadhyay K (2011) Purification and preliminary characterization of xylanase from Thermomyces lanuginosus strain SS-8. 3 Biotech 1:255–259

    Article  Google Scholar 

  • Silva JC, Denny R, Dorschel CA, Gorenstein M, Kass IJ, Li GZ et al (2005) Quantitative proteomic analysis by accurate mass retention time pairs. Anal Chem 77:2187–2200

    Article  CAS  Google Scholar 

  • Singh S, Pillay B, Prior BA (2000) Thermal stability of β-xylanases produced by different Thermomyces lanuginosus strains. Enzyme Microb Technol 26:502–508

    Article  CAS  Google Scholar 

  • Singh S, Mandala AM, Prior BA (2003) Thermomyces lanuginosus: properties of strains and their hemicellulases. FEMS Microbiol Rev 27:3–16

    Article  CAS  Google Scholar 

  • Sriyapai T, Somyoonsap P, Matsui K, Kawai F, Chansiri K (2011) Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris. J Biosci Bioeng 111:528–536

    Article  CAS  Google Scholar 

  • The Quiaexpressionist (2003) A handbook of high-level expression and purification of 6× His-tagged proteins, 5th edn. Quiagen, Valencia, pp 39–40

    Google Scholar 

  • Torronen A, Rouvinen J (1997) Structural and functional properties of low molecular weight endo-1,4-β-xylanases. J Biotechnol 57:137–149

    Article  CAS  Google Scholar 

  • Verma D, Satyanarayana T (2012) Cloning, expression and applicability of thermo-alkali-stable xylanase of Geobacillus thermoleovorans in generating xylooligosaccharides from agro-residues. Bioresour Technol 107:333–338

    Article  CAS  Google Scholar 

  • Wakarchuk WW, Campbell RL, Sung WL, Davoodi J, Yaguchi M (1994) Mutational and crystallographic analyses of the active-site residues of Bacillus cirlulans xyalanase. Protein Sci 3:467–475

    Article  CAS  Google Scholar 

  • Wang YR, Zhang HL, He YZ, Luo HY, Yao B (2007) Characterization, gene cloning, and expression of a novel xylanase XYNB from Streptomyces olivaceoviridis A1. Aquaculture 267:328–334

    Article  CAS  Google Scholar 

  • Wang Y, Fu Z, Huang H, Zhang H, Yao B, Xiong H et al (2012) Improved thermal performance of Thermomyces lanuginosus GH11 xylanase by engineering of an Nterminal disulfide bridge. Bioresour Technol 112:275–279

    Article  CAS  Google Scholar 

  • Xue GP, Denman SE, Glassop D, Johnson JS, Dierens LM, Gobius KS, Anyward JH (1995) Modification of a xylanase cDNA isolated from an anaerobic fungus Neocallimastix patriciarum for high-level expression in Escherichia coli. J Biotechnol 38:269–277

    Article  CAS  Google Scholar 

  • Zhang C, Jia L, Wanga S, Qu J, Li K, Xu L, Shi Y, Yan Y (2010) Biodegradation of β-cypermethrin by two Serratia sp. with different cell surface hydrophobicity. Bioresour Technol 101:3423–3429

    Article  CAS  Google Scholar 

  • Zhou C, Li D, Wu M, Wang W (2008) Optimized expression of an acid xylanase from Aspergillus usamii in Pichia pastoris and its biochemical characterization. World J Microbiol Biotechnol 24:1393–1401

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge the Government of Jharkhand, Department of Agriculture for providing infrastructure development fund to Department of Biotechnology, Birla Institute of Technology, Mesra, Ranchi for financial supports. Smriti Shrivastava is thankful to Council of Scientific and Industrial Research for Senior Research Fellowship (Grant No: 09/554(0017)/2008 EMR-I). Authors also acknowledge Central Instrumentation Facility at Birla Institute of Technology, Mesra, India for providing Scanning electron microscopy facility and Shalini Jalan and Aarushi Sharma (B. Tech Biotechnology students at Birla institute of Technology, Mesra, Ranchi, Jharkhand, Batch 2007–2011) for performing SEM analysis.

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  1. Pratyoosh Shukla

    Present address: Department of Microbiology, M.D. University, Rohtak, 124001, India

Authors and Affiliations

  1. Department of Biotechnology, Birla Institute of Technology, Mesra, Ranchi, 835215, India

    Smriti Shrivastava, Pratyoosh Shukla & Kunal Mukhopadhyay

  2. Waters India Pvt Ltd, 36A, II Phase, Peenya Industrial Area, Bangalore, 560058, India

    Putchen Dakshinamoorthy Deepalakshmi

Authors
  1. Smriti Shrivastava
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  2. Pratyoosh Shukla
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  3. Putchen Dakshinamoorthy Deepalakshmi
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  4. Kunal Mukhopadhyay
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Correspondence to Pratyoosh Shukla.

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Shrivastava, S., Shukla, P., Deepalakshmi, P.D. et al. Characterization, cloning and functional expression of novel xylanase from Thermomyces lanuginosus SS-8 isolated from self-heating plant wreckage material. World J Microbiol Biotechnol 29, 2407–2415 (2013). https://doi.org/10.1007/s11274-013-1409-y

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  • DOI: https://doi.org/10.1007/s11274-013-1409-y

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