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A novel thermostable xylanase of Paenibacillus macerans IIPSP3 isolated from the termite gut

  • Pratibha Dheeran
  • N. Nandhagopal
  • Sachin Kumar
  • Yogesh K. Jaiswal
  • Dilip K. Adhikari
Bioenergy/Biofuels/Biochemicals

Abstract

Xylanase is an enzyme in high demand for various industrial applications, such as those in the biofuel and pulp and paper fields. In this study, xylanase-producing microbes were isolated from the gut of the wood-feeding termite at 50°C. The isolated microbe produced thermostable xylanase that was active over a broad range of temperatures (40–90°C) and pH (3.5–9.5), with optimum activity (4,170 ± 23.5 U mg−1) at 60°C and pH 4.5. The enzyme was purified using a strong cation exchanger and gel filtration chromatography, revealing that the protein has a molecular mass of 205 kDa and calculated pI of 5.38. The half-life of xylanase was 6 h at 60°C and 2 h at 90°C. The isolated thermostable xylanase differed from other xylanases reported to date in terms of size, structure, and mode of action. The novelty of this enzyme lies in its high specific activity and stability at broad ranges of temperature and pH. These properties suggest that this enzyme could be utilized in bioethanol production as well as in the paper and pulp industry.

Keywords

Thermostable xylanase Thermophiles Termite gut Paenibacillus macerans IIPSP3 Enzyme purification 

Notes

Acknowledgments

All of the authors thank Dr M.O. Garg, Director IIP, Dehradun for his valuable suggestions and encouragement to carry out this research and also acknowledge their thanks to TCGA, New Delhi for the internal amino acid sequencing using LC–MS/MS. One of the authors gratefully acknowledges a Senior Research Fellowship awarded by the Council of Scientific and Industrial Research (CSIR), India.

References

  1. 1.
    Anonymous (2007) Termite guts may yield novel enzymes for better biofuel production. In: ScienceDaily 25 Nov. Available at: http://www.sciencedaily.com-/releases/2007/11/071121145002.htm. Accessed: 21 Apr 2010
  2. 2.
    Anonymous (2007) Biofuels: bringing biological solutions to energy challenges. US Department of Energy Office of Science, Washington D.C. Google Scholar
  3. 3.
    Bataillon M, Cardinali APN, Castillon N, Duchiron F (2000) Purification and characterization of a moderately thermostable xylanase from Bacillus sp. strain SPS-0. Enzyme Microb Technol 26:187–192PubMedCrossRefGoogle Scholar
  4. 4.
    Beg QK, Kapoor M, Mahajan L, Hoondal GS (2001) Microbial xylanases and their industrial applications: a review. Appl Microbiol Biotechnol 56:326–338PubMedCrossRefGoogle Scholar
  5. 5.
    Biely P (1985) Microbial xylanolytic systems. Trends Biotechnol 3:286–290CrossRefGoogle Scholar
  6. 6.
    Breccia JD, Sineriz F, Baigori MD, Castro GR, Hatti-Kaul R (1998) Purification and characterization of a thermostable xylanase from Bacillus amyloliquefaciens. Enzyme Microb Technol 22:42–49CrossRefGoogle Scholar
  7. 7.
    Brennan YL, Callen WN, Christoffersen L, Dupree P, Goubet F, Healey S et al (2004) Unusual microbial xylanases from insect guts. App Environ Microbiol 70:3609–3617CrossRefGoogle Scholar
  8. 8.
    Brune A (2007) Woodworker’s digest. Nature 450:487–488PubMedCrossRefGoogle Scholar
  9. 9.
    Cho MC, Bai S (1997) Purification and characterization of xylanase from Bacillus sp. strain DSNC 101. J Microbiol Biotechnol 7:386–390Google Scholar
  10. 10.
    Collins T, Gerday C, Feller G (2005) Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 29:3–23PubMedCrossRefGoogle Scholar
  11. 11.
    Cordeiro CAM, Martins MLL, Luciano AB, da Silva RF (2002) Production and properties of xylanase from thermophilic Bacillus sp. Braz Arch Biol Technol 45:413–418CrossRefGoogle Scholar
  12. 12.
    Dheeran P, Kumar S, Jaiswal YK, Adhikari DK (2010) Characterization of hyperthermostable α-amylase from Geobacillus sp. IIPTN. Appl Microbiol Biotechnol 86:1857–1866PubMedCrossRefGoogle Scholar
  13. 13.
    Ikai AJ (1980) Thermostability and aliphatic index of globular proteins. J Biochem 88:1895–1898PubMedGoogle Scholar
  14. 14.
    Johnvesly B, Virupakshi S, Patil GN, Ramalingam A, Naik GR (2002) Cellulase-free thermostable alkaline xylanase from thermophilic and alkalophilic Bacillus sp. JB-99. J Microbiol Biotechnol 12:153–156Google Scholar
  15. 15.
    Ko CH, Lin ZP, Tu J, Tsai CH, Liu CC, Chen HT, Wang TP (2010) Xylanase production by Paenibacillus campinasensis BL11 and its pretreatment of hardwood kraft pulp bleaching. Int Biodet Biodeg 64:13–19CrossRefGoogle Scholar
  16. 16.
    Ko CH, Tsai CH, Tu J, Yang BY, Hsieh DL, Jane WN, Shih TL (2011) Identification of Paenibacillus sp. 2S–6 and application of its xylanase on biobleaching. Int Biodet Biodeg 65:334–339CrossRefGoogle Scholar
  17. 17.
    Kulkarni N, Shendye A, Rao M (1999) Molecular and biotechnological aspects of xylanases. FEMS Microbiol Rev 23:411–456PubMedCrossRefGoogle Scholar
  18. 18.
    Kumar S, Singh SP, Mishra IM, Adhikari DK (2009a) Ethanol and xylitol production from glucose and xylose at high temperature by Kluyveromyces sp. IIPE453. J Ind Microbiol Biotechnol 36:1483–1489PubMedCrossRefGoogle Scholar
  19. 19.
    Kumar S, Singh SP, Mishra IM, Adhikari DK (2009b) Recent advances in production of bioethanol from lignocellulosic biomass. Chem Eng Technol 32:517–526CrossRefGoogle Scholar
  20. 20.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  21. 21.
    Lee TH, Lim PO, Lee YE (2007) Cloning characterization and expression of Xylanase A gene from Paenibacillus sp. DG-22 in Escherichia coli. J Microbiol Biotechnol 17:29–36PubMedGoogle Scholar
  22. 22.
    Lin LL, Thomson JA (1991) An analysis of the extracellular xylanases and cellulases of Butyrivibrio fibrisolvens H17c. FEMS Microbiol Lett 84:197–204CrossRefGoogle Scholar
  23. 23.
    Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666CrossRefGoogle Scholar
  24. 24.
    Liu CJ, Suzuki T, Hirata S, Kawai K (2003) The processing of high-molecular-weight Xylanase (XynE, 110 kDa) from Aeromonas caviae ME-l to 60-kDa xylanase (XynE60) in Escherichia coli and purification and characterization of XynE60. J Biosci Bioeng 95:95–101PubMedGoogle Scholar
  25. 25.
    Lopez-Fernandez CL, Rodriguez J, Ball AS, Copa-Patino JL, Perez-Leblic MI, Arias ME (1998) Application of the affinity binding of xylanases to oat-spelt xylan in the purification of endoxylanase CM-2 from Streptomyces chattanoogensis CECT 3336. Appl Microbiol Biotechnol 50:284–287CrossRefGoogle Scholar
  26. 26.
    Lowry OH, Rosebrough AJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–273PubMedGoogle Scholar
  27. 27.
    Matson E, Ottesen E, Leadbetter J (2007) Extracting DNA from the gut microbes of the termite (Zootermopsis nevadensis). J Vis Exp 4:195PubMedGoogle Scholar
  28. 28.
    Matsui T, Tokuda G, Shinzato N (2009) Termites as functional gene resources. Recent Patents Biotechnol 3:10–18CrossRefGoogle Scholar
  29. 29.
    Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugars. Anal Chem 31:426–428CrossRefGoogle Scholar
  30. 30.
    Ninawe S, Kuhad RC (2005) Use of xylan-rich cost effective agro-residues in the production of xylanase by Streptomyces cyaneus SN32. J Appl Microbiol 99:1141–1148PubMedCrossRefGoogle Scholar
  31. 31.
    Pason P, Kyu KL, Ratanakhanokchai K (2006) Paenibacillus curdlanolyticus strain B-6 xylanolytic—ellulolytic enzyme system that degrades insoluble polysaccharides. Appl Environ Microbiol 72:2483–2490PubMedCrossRefGoogle Scholar
  32. 32.
    Schäfer A, Konrad R, Kuhnigk T, Kämpfer P, Herte H, König H (1996) Hemicellulose-degrading bacteria and yeasts from the termite gut. J Appl Microbiol 80:471–478CrossRefGoogle Scholar
  33. 33.
    Shi P, Tian J, Yuan T, Liu X, Huang H, Bai Y, Yang P, Chen X, Wu N, Yao B (2010) Paenibacillus sp. strain E18 bifunctional xylanase-glucanase with a single catalytic domain. Appl Environ Microbiol 76:3620–3624PubMedCrossRefGoogle Scholar
  34. 34.
    Varma A, Kollia BK, Paula J, Saxenaa S, König H (1994) Lignocellulose degradation by microorganisms from termite hills and termite guts: a survey on the present state of art. FEMS Microbiol Rev 15:9–28CrossRefGoogle Scholar
  35. 35.
    Wang CY, Chan H, Lin HT, Shyu YT (2010) Production, purification and characterization of a novel halostable xylanase from Bacillus sp. NTU-06. Ann Appl Biol 156:187–197CrossRefGoogle Scholar
  36. 36.
    Warnecke F, Luginbühl P, Ivanova N, Ghassemian M, Richardson TH, Stege JT et al (2007) Metagenomic and functional analysis of hindgut microbiota of a wood-feeding higher termite. Nature 450:560–565PubMedCrossRefGoogle Scholar
  37. 37.
    Yin LJ, Lin HH, Chiang YI, Jiang ST (2010) Bioproperties and purification of xylanase from Bacillus sp. YJ6. J Agric Food Chem 58:557–562PubMedCrossRefGoogle Scholar

Copyright information

© Society for Industrial Microbiology and Biotechnology 2012

Authors and Affiliations

  • Pratibha Dheeran
    • 1
  • N. Nandhagopal
    • 1
  • Sachin Kumar
    • 1
    • 2
  • Yogesh K. Jaiswal
    • 3
  • Dilip K. Adhikari
    • 1
  1. 1.Biotechnology AreaIndian Institute of PetroleumDehradunIndia
  2. 2.Sardar Swaran Singh National Institute of Renewable EnergyKapurthalaIndia
  3. 3.School of Studies in BiochemistryJiwaji UniversityGwaliorIndia

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