Skip to main content
SpringerLink
Log in

Characterization of a recombinant thermostable xylanase from deep-sea thermophilic Geobacillus sp. MT-1 in East Pacific

  • Biotechnologically Relevant Enzymes and Proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A novel xylanase-producing thermophilic strain MT-1 was isolated from a deep-sea hydrothermal field in east Pacific. A xylanase gene encoding 331 amino-acid peptide from this isolate was cloned and expressed in Escherichiacoli. The recombinant xylanase exhibited maximum activity at 70°C and had an optimum pH of 7.0. It was active up to 90°C and showed activity over a wide pH ranging from 5.5 to 10.0. The crude xylanase presented similar properties in temperature and pH to those of the recombinant xylanase. The recombinant xylanase was stable in 1 mM of enzyme inhibitors (PMSF, EDTA, 2-ME or DTT) and in 0.1% detergents (Tween 20, Chaps or Triton X-100), whereas, it was strongly inhibited by sodium dodecyl sulfate (SDS) (1 mM). In addition, its catalytic function was stable in the presence of Li+, Na+ or K+. However, it was strongly inhibited by Ni2+, Mn2+, Co2+, Cu2+, Zn2+, Cd2+, Hg2+ and Al3+ (1 or 0.1 mM). The K m and V max of the recombinant xylanase for oat spelt xylan were calculated to be 1.579 mg/ml and 289 μmol/(min • mg), respectively. Our study, therefore, presented a rapid overexpression and purification of xylanase from deep-sea thermophile aimed at improving the enzyme yield for industrial applications and scientific research.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Bailey MJ, Biely P, Poutanen K (1992) Interlaboratory testing of methods for assay of xylanase activity. J Biotechnol 23:257–270

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Bergquist PL, Gibbs MD, Morris DD, TeÓ VSJ, Saul DJ, Morgan HW (1999) Molecular diversity of thermophilic cellulolytic and hemicellulolytic bacteria. FEMS Microbiol Ecol 28:99–110

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Blanco A, Díaz P, Zueco J, Parascandola P, Pastor FIJ (1999) A multidomain xylanase from a Bacillus sp. with a region homologous to thermostabilizing domains of thermophilic enzymes. Microbiology 145:2163–2170

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Christakopoulos P, Katapodis P, Kalogeris E, Kekos D, Macris BJ, Stamatis H, Skaltsa H (2003) Antimicrobial activity of acidic xylooligosaccharides produced by family 10 and 11 endoxylanases. Int J Biol Macromol 31:171–175

    Article  CAS  Google Scholar 

  • Clarke JH, Davidson K, Gilbert HJ, Fontes C, Hazlewood GP (1996) A modular xylanase from mesophilic Cellulomonasfimi contains the same cellulose-binding and thermostabilizing domains as xylanases from thermophilic bacteria. FEMS Microbiol Lett 139:27–35

    Article  CAS  Google Scholar 

  • Connerton I, Cumming N, Harris GW, Debeire P, Breton C (1999) A single domain thermophilic xylanase can bind insoluble xylan: evidence for surface aromatic clusters. Biochim Biophys Acta 1433:110–121

    Article  CAS  Google Scholar 

  • Fontes CM, Hazlewood GP, Morag E, Hall J, Hirst BH, Gilbert HJ (1995). Evidence for a general role for non-catalytic thermostabilizing domains in xylanases from thermophilic bacteria. Biochem J 307:151–158

    Article  CAS  Google Scholar 

  • Gibbs MD, Reeves RA, Bergquist PL (1995) Cloning sequencing and expression of a xylanase Gene from the extreme thermophile Dictyoglomusthermophilum Rt46B.1 and activity of the enzyme on fiber-bound substrate. Appl Environ Microbiol 61:4403–4408

    Article  CAS  Google Scholar 

  • Gilkes NR, Henrissat B, Kilburn DG, Miller RC, Warren RAJ (1991) Domains in microbial β-1,4-glycanases; sequence conservation, function, and enzyme families. Microbiol Rev 55:303–315

    CAS  PubMed  PubMed Central  Google Scholar 

  • Khanna S, Gauri P (1993) Regulation, purification and properties of xylanase from Cellulomonasfimi. Enzyme Microb Technol 15:990–995

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 57:685

    Google Scholar 

  • Martínez MA, Delgado OD, Baigorí MD, Siòeriz F (2005) Sequence analysis, cloning and over-expression of an endoxylanase from the alkaliphilic Bacillushalodurans. Biotechnol Lett 27:545–550

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Oppenheimer CH, ZoBell CE (1952) The growth and viability of sixty-three species of marine bacteria as influenced by hydrostatic pressure. J Mar Res 11:10–18

    Google Scholar 

  • Shah AR, Madamwar D (2005) Xylanase production by a newly isolated Aspergillusfoetidus strain and its characterization. Process Biochem 40:1763–1771

    Article  CAS  Google Scholar 

  • Shibuya H, Kaneko S, Hayashi K (2000) Enhancement of the thermostability and hydrolytic activity of xylanase by random gene shuffling. Biochem J 349:651–656

    Article  CAS  Google Scholar 

  • Subramaniyan S, Prema P (2002) Biotechnology of microbial xylanases: enzymology, molecular biology, and application. Crit Rev Biotechnol 22:33–64

    Article  CAS  Google Scholar 

  • Sunna A, Antranikian G (1997) Xylanolytic enzymes from fungi and bacteria. Crit Rev Biotechnol 17:39–67

    Article  CAS  Google Scholar 

  • Sunna A, Gibbs MD, Bergquist PL (2000) A novel thermostable multidomain 1,4-β-xylanase from ‘Caldibacilluscellulovorans’ and effect of its xylan-binding domain on enzyme activity. Microbiology 146:2947–2955

    Article  CAS  Google Scholar 

  • Viikari L, Kantelineo A, Bundquist J, Linko M (1994) Xylanase in bleaching: from an idea to the industry. FEMS Microbiol Rev 13:335–350

    Article  CAS  Google Scholar 

  • Viikari L, Sundquist J, Kettunen J (1991) Xylanase enzymes promote pulp bleaching. Pap Puu (Pap Tim) 73:384–389

    CAS  Google Scholar 

  • Wong KY, Tan LUL, Saddler JN (1988) Multiplicity of β-1,4-xylanase in microorganisms: functions and applications. Microbiol Rev 52:305–317

    CAS  PubMed  PubMed Central  Google Scholar 

  • Xiaoyan W, Jianyi S (2005) Construction, expression and characterization of a thermostable xylanase. Curr Microbiol 51:188–192

    Article  Google Scholar 

Download references

Acknowledgements

This work was financially supported by the China Ocean Mineral Resources R & D Association (DY105-02-04-05) and the National Natural Science Foundation of China (40576076).

Author information

Authors and Affiliations

  1. School of Life Sciences, Xiamen University, Xiamen 361005, People’s Republic of China

    Suijie Wu

  2. Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, 178 Daxue Rd., Xiamen 361005, People’s Republic of China

    Suijie Wu, Bin Liu & Xiaobo Zhang

Authors
  1. Suijie Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bin Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaobo Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiaobo Zhang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, S., Liu, B. & Zhang, X. Characterization of a recombinant thermostable xylanase from deep-sea thermophilic Geobacillus sp. MT-1 in East Pacific. Appl Microbiol Biotechnol 72, 1210–1216 (2006). https://doi.org/10.1007/s00253-006-0416-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-006-0416-4

Keywords

Search

Navigation