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Alkali-thermostable and cellulase-free xylanase production by an extreme thermophile Geobacillus thermoleovorans

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The optimization of cultural variables resulted in a marked enhancement in the secretion of cellulase-free and alkali-thermostable xylanase (EC by an extreme thermophile Geobacillus thermoleovorans. The enzyme secretion was enhanced when the medium was supplemented with xylan (0.15%) and Tween-80 (0.1% v/v). In wheat bran-tryptone medium, the peak in enzyme production was attained within 42 h in a fermenter as compared to 72 h in shake flasks. Optimization of the culture conditions resulted in a 7.72-fold enhancement in enzyme production. The cellulase-free xylanase was optimally active at pH 8.5 and 80°C, and it was found to be useful in the pre-bleaching process of paper pulps.

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  1. Adamsen AK, Lindhagen J, Ahring BK (1995) Optimization of extracellular xylanase production by Dictyoglomus sp. B1 in continuous culture. Appl Microbiol Biotechnol 44:327–332

  2. Archana A, Satyanarayana T (1997) Xylanase production by thermophilic Bacillus licheniformis A99 in solid-state fermentation. Enzyme Microb Technol 21:12–17

  3. Archana A, Satyanarayana T (1998) Cellulase-free xylanase production by thermophilic Bacillus licheniformis A99. Indian J Microbiol 38:135–139

  4. Babu KR, Satyanarayana T (1993) Extracellular calcium-inhibited alpha-amylase of Bacillus coagulans B49. Enzyme Microb Technol 15:1066–1069

  5. Bailey MJ, Buchert J, Viikari L (1993) Effect of pH on production of xylanase by Trichoderma reesei on xylan and cellulose-based media. Appl Microbiol Biotechnol 40:224–229

  6. Bartholomew WH, Karrow EO, Sfat MR, Wilhelm RH (1950) Oxygen transfer in submerged fermentations of Streptomyces griseus. Indust Eng Chem 42:1801–1809

  7. Bastawde KB (1992) Xylan structure, microbial xylanases, and their mode of action. World J Microbiol Biotechnol 8:353–368

  8. Beg QK, Bhushan B, Kapoor M, Hoondal G (2000) Production and characterization of thermostable xylanase and pectinase from Streptomyces sp. QG-11-3. J Indust Microbiol Biotechnol 24:396–402

  9. Biely P, Hirch J, la Grange DC, Van Zyl WH, Prior BA (2000) A chromogenic substrate for β-xylosidase-coupled assay of α-glucuronidase. Ann Biochem 286:289–294

  10. Breccia JD, Sifieriz F, Baigori MD, Castro GR, Hatti-Kaul R (1998) Purification and characterization of a thermostable xylanase from Bacillus amyloliquefaciens. Enzyme Microb Technol 22:42–49

  11. Castenholz RW (1979) In: Shilo M (ed) Strategies of microbial life in extreme environments. Science Research Report 13. Dahlem Konferenzen, Berlin, pp 373–392

  12. Damiano VB, Bocchini DA, Gomes E, Da Silva R (2003) Application of crude xylanase from Bacillus licheniformis 77-2 to the bleaching of eucalyptus Kraft pulp. World J Microbiol Biotechnol 19:139–144

  13. Demain ACR (1972) Influence of environment on the control of enzyme synthesis. J Appl Chem Biotechnol 22:245–259

  14. Dhillon A, Gupta JK, Khanna S (2000) Enhanced production, purification and characterization of a novel cellulase-poor, thermostable alkalitolerant xylanase from Bacillus circulans AB16. Process Biochem 35:210–215

  15. Esteban R, Villanueva JR, Villa TG (1982) β-D-xylanases of Bacillus circulans WL-12. Can J Microbiol 28:733–739

  16. Gomes DJ, Gomes J, Steiner W (1994) Factors influencing the induction of endoxylanases by Thermoascus aurantiacus. J Biotechnol 33:87–94

  17. Gruninger H, Fiechter A (1986) A novel highly thermostable D-xylanase. Enzyme Microb Technol 8:309–314

  18. Gübitz GM, Haltrich D, Latal B, Steiner W (1997) Mode of depolymerization of hemicellulose by various mannanases and xylanases in relation to their ability to bleach softwood pulp. Appl Microbiol Biotechnol 47:658–662

  19. Humphrey A (1998) Shake-flask to fermenter, what have we learnt? Biotechnol Prog 14:3–7

  20. Illias M, Hoq MM (1998) Effect of agitation rate on growth and production of cellulase-free xylanase by Thermomyces lanuginosus MH4 in a bioreactor. World J Microbiol Biotechnol 14:765–767

  21. Kamra P, Satyanarayana T (2004) Xylanase production by the thermophilic mold Humicola lanuginosa in solid-state fermentation. Appl Biochem Biotechnol 119:145–157

  22. Kantelinen A, Rantanen T, Buchert J, Vikari L (1993) Enzymatic solubilization of fiber-bound and isolated birch xylans. J Biotechnol 28:219–228

  23. Karlsson EN, Dahlberg L, Jorto N Gorton L, Holst H (1998) Enzymatic specificity and hydrolysis pattern of the catalytic domain of the xylanase (Xynl) from Rhodothermus marinus. J Biotechnol 60:23–35

  24. Kaupinen T, Teschke K, Savela. A, Kogevinas M, Boffetta P (1997) International data base of exposure measurement in the pulp, paper and paper product industries. Int Arch Occup Environ Health 70:119–127

  25. Kaur P, Satyanarayana T (2004) Production and saccharification by a thermostable and neutral glucoamylase of thermophilic mould Thermomucor indicae-seudaticae. World J Microbiol Biotechnol 20:419–425

  26. Khasin A, Alchanati I, Shoham Y (1993) Purification and characterization of a thermostable xylanase from Bacillus stearothermophilus T-6. Appl Environ Microbiol 59:1725–1730

  27. Kulkarni N, Rao M (1996) Application of xylanases from alkaliphilic thermophilic Bacillus sp. NCIM 59: biobleaching of baggasse pulp. J Biotechnol 13:151–172

  28. Kulkarni N, Shendye A, Rao M (1999) Molecular and Biological aspects of xylanases. FEMS Microbiol Rev 23:411–456

  29. Levin L, Forschiassin F (1998) Influence of growth conditions on the production of xylanolytic enzymes by Trametes trogii. World J Microbiol Biotechnol 14:443–446

  30. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin-phenol reagent. J Biol Chem 193:265–275

  31. Mamo G, Gessesse A (2000) Immobilization and alkaliphilic Bacillus sp. cells for xylanase production using batch and continuous culture. Appl Biochem Biotechnol 87:95–101

  32. Mandels M, Hontz L, Nystrom J (1974) Enzymatic hydrolysis of waste cellulose. Biotechnol Bioeng 16:1471–1493

  33. Marques S, Alves L, Ribeiros GE, Collaco MTA (1998) Characterization of a thermotolerant and alkalitolerant xylanase from a Bacillus sp. Appl Biochem Biotechnol 73:159–171

  34. Mathrani IM, Ahring BK (1992) Thermophilic and alkaliphilic xylanase from several Dictyoglomus isolates. Appl Microbiol Biotechnol 38:23–27

  35. Meyrath J, Suchanek G (1972) Inoculation techniques–effects due to quality and quantity of inoculum. In: Norris Jr, Ribbons DW (eds) Methods in microbiology, vol 7B. Academic Press, London, pp 159–209

  36. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugars. Anal Chem 31:426–428

  37. Moon SH, Parulekar SJ (1991) A parametric study of protease production in batch and fed batch cultures of Bacillus firmus. Biotechnol Bioeng 37:467–483

  38. Narang S, Satyanarayana T (2001) Thermostable α-amylase production by an extreme thermophile Bacillus thermoleovorans. Lett Appl Microbiol 32:31–35

  39. Okazaki W, Akiba T, Horikoshi K, Akahoshi R (1985) Purification and characterization of xylanases from alkaliphilic thermophilic Bacillus spp. Agric Biol Chem 49:2033–2039

  40. Prema P (2005) Xylanases. In: Pandey A, Webb C, Soccol CR, Larroche C (eds) Enzyme technology. Asiatech Publ, New Delhi, pp 333–346

  41. Priest FG (1977) Extracellular enzyme synthesis in the genus Bacillus. Bacteriol Rev 41:711–753

  42. Saraswat V, Bisaria VS (1997) Biosynthesis of xylanolytic and xylan-debranching enzymes in Melanocarpus albomyces 11S 68. J Ferment Bioeng 83:352–357

  43. Sen S, Satyanarayana T (1993) Optimization of alkaline protease production by thermophilic Bacillus licheniformis S-40. Indian J Microbiol 33:43–47

  44. Shaku M, Koike S, Udaka S (1980) Cultural conditions for protein production by Bacillus brevis No. 47. Agric Biol Chem 44:99–103

  45. Sunna A, Prowe SG, Stoffregen T, Antranikian G (1997) Characterization of the xylanases from the newly isolated thermophilic xylan degrading Bacillus thermoleovorans strain K-3d and Bacillus flavothermus strain LB3A. FEMS Microbiol Lett 148:209–216

  46. TAPPI (1992) Kappa number of pulp. T-236 CM-85. In: TAPPI Test Methods (1992–93). Technical Association of the Pulp and Paper Industry, Atlanta

  47. Techapun C, Poosaran N, Watanabe M, Sasaki K (2003) Thermostable and alkaline-tolerant microbial cellulase-free xylanases produced from agricultural wastes and the properties required for use in pulp bleaching bioprocesses: a review. Process Biochem 38:1327–1340

  48. Torres AL, Roncero MB, Colom JF, Pastor FIJ, Blanco A, Vidal T (2000) Effect of a novel enzyme on fibre morphology during ECF bleaching of oxygen delignified Eucalyptus kraft pulps. Bioresource Technology 74:135–140

  49. Tuohy MG, Buckley RJ, Griffin TO, Connelly IC, Shanley NA, Filho EXF (1989) Enzyme production by solid-state cultures of aerobic fungi on lignocellulosic substrates. In: Coughlan MP (ed) Enzyme systems for lignocellulose degradation. Elsevier, London, pp 293–312

  50. Uma Maheswar Rao JL, Satyanarayana T (2003) Enhanced secretion and low temperature stabilization of a hyperthermostable and Ca2+-independent α-amylase of Geobacillus thermoleovorans by surfactants. Lett Appl Microbiol 36:191–196

  51. Viikari L, Ranua M, Kantelinen A, Sunduist J, Linko M (1986) Bleaching with enzymes. In: Proc 3rd Int Conf Biotechnology in the Pulp and Paper Industry. STFI, Stockholm, Sweden, pp 67–69

  52. Vyas P, Chauthaiwale V, Phadatare S, Deshpande V, Srinivasan MC (1990) Studies on the alkaliphilic Streptomyces with extracellular xylanolytic activity. Biotechnol Lett 12:225–228

  53. Yin YL, Baidoo SK, Jin LZ, Liu YG, Schulze H, Simmins PH (2001) The effect of different carbohydrate and protease supplementation on apparent (ideal and overall) digestibility of nutrients of five hulless barley varieties in young pigs. Livest Prod Sci 71:109–120

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AS and SK are grateful to the University Grants Commission (UGC), Government of India for providing financial assistance during the course of this investigation.

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Correspondence to T. Satyanarayana.

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Sharma, A., Adhikari, S. & Satyanarayana, T. Alkali-thermostable and cellulase-free xylanase production by an extreme thermophile Geobacillus thermoleovorans . World J Microbiol Biotechnol 23, 483–490 (2007).

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  • Cellulase-free xylanase
  • Extreme thermophile
  • Geobacillus thermoleovorans
  • Optimization
  • Paper pulp
  • Pre-bleaching