Skip to main content

Advertisement

SpringerLink
Log in

Cloning, expression and characterization of xylose isomerase, XylA, from Caldanaerobacter subterraneus subsp. yonseiensis

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

The xylA gene, coding for xylose isomerase, from the extreme thermophile, Caldanaerobacter subterraneus subsp. yonseiensis was cloned, sequenced, and expressed in Escherichia coli. The nucleotide sequence of the xylA gene encoded a polypeptide of 438 residues with a calculated molecular weight of 50,170 Da. The purified XylA showed high sequence homology (92% identity) with that of Thermoanaerobacter thermohydrosulfuricus. The recombinant enzyme expressed in Escherichia coli was purified by heat treatment and gel chromatography. The purified enzyme was thermostable with optimal activity at 95°C. The enzyme required divalent cations including Zn2+ for its maximal activity and thermostability.

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

Similar content being viewed by others

References

  • Antrim RL, Colilla W, Schnyder BJ (1979) Glucose isomerase production of high-fructose syrups. In: Wingard LB (ed) Applied biochemistry and bioengineering. Academic Press, New York, pp 97–155

    Google Scholar 

  • Bhosale SH, Rao MB, Deshpande VV (1996) Molecular and industrial aspects of glucose isomerase. Microbiol Rev 60:280–300

    CAS  PubMed  Google Scholar 

  • Brown SH, SjVholm C, Kelly RM (1993) Purification and characterization of a highly thermostable glucose isomerase produced by the extremely thermophilic eubacterium, Thermotoga maritima. Biotechnol Bioeng 41:878–886

    Article  CAS  PubMed  Google Scholar 

  • Cha J, Cho Y, Whitaker RD, Carrell HL, Glusker JP, Karplus PA, Batt CA (1994) Perturbing the metal site in XylA. Effect of mutations of His-220 on enzyme stability. J Biol Chem 269:2687–2694

    CAS  PubMed  Google Scholar 

  • Chandrakant P, Bisaria VS (1998) Simultaneous bioconversion of cellulose and hemicellulose to ethanol. Crit Rev Biotechnol 18:295–331

    Article  CAS  PubMed  Google Scholar 

  • Dekker K, Yamagata H, Sakaguchi K, Ucaka S (1991) Xylose(Glucose) isomerase gene from the thermophile Clostridium thermohydrosulfuricum; cloning, sequencing, and expression in Escherichia coli. Agric Biol Chem 55:221–227

    CAS  PubMed  Google Scholar 

  • Dische Z, Borefreund E (1951) A new spectrophotometric method for the detection and determination of keto sugars and trioses. J Biol Chem 192:583–587

    CAS  PubMed  Google Scholar 

  • Epting KL, Vieille C, Zeikus JG, Kelly RM (2005) Influence of divalent cations on the structural thermostability and thermal inactivation kinetics of class II XylAs. FEBS J 272:1454–1464

    Article  CAS  PubMed  Google Scholar 

  • Erbeznik M, Dawson KA, Strobel HJ (1998) Cloning and characterization of transcription of xylAB operon in Thermoanacerobacter ethanolicus. J Bacteriol 18:1103–1109

    Google Scholar 

  • Fardeau ML, Salinas MB, L’Haridon S, Jeanthon C, Verhé F, Cayol JL, Patel BKC, Garcia JL, Ollivier B (2004) Isolation from oil reservoirs of novel thermophilic anaerobes phylogenetically related to Thermoanaerobacter subterraneus: reassignment of T. subterraneus, Thermoanaerobacter yonseiensis, Thermoanaerobacter tengcongensis and Carboxydibrachium pacificum to Caldanaerobacter subterraneus gen. nov., sp. nov., comb. nov. as four novel subspecies. Int J Syst Evol Microbiol 54:467–474

    Article  CAS  PubMed  Google Scholar 

  • Goldberg JB, Ohman DE (1984) Cloning and expression in Pseudomonas aeruginosa of a gene involved in the production of alginate. J Bacteriol 158:1115–1121

    CAS  PubMed  Google Scholar 

  • Hall TA (1990) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98

    Google Scholar 

  • Kim BC, Grote R, Lee DW, Antranikian G, Pyun YR (2001) Thermoanaerobacter yonseiensis sp. nov., a novel extremely thermophilic, xylose-utilizing bacterium that grows at up to 85°C. Int J Syst Evol Microbiol 51:1539–1548

    CAS  PubMed  Google Scholar 

  • Kuyper M, Hartog MMP, Toirkens MJ, Almering MJH, Winkler AA, Dijken JPV, Pronk JT (2005) Metabolic engineering of a xylose-isomerase-expressing Saccharomyces cerevisiae strain for rapid anaerobic xylose fermentation. FEMS Yeast Res 5:399–409

    Article  CAS  PubMed  Google Scholar 

  • Liu SY, Wiegel J, Gherardini FC (1996) Purification and cloning of a thermostable xylose (glucose) isomerase with an acidic pH optimum from Thermoanaerobacterium strain JW/SL-YS 489. J Bacteriol 178:5938–5945

    CAS  PubMed  Google Scholar 

  • Vieille C, Hess JM, Kelly RM, Zeikus JG (1995) xylA cloning and sequencing and biochemical characterization of XylA from Thermotoga neapolitana. Appl Environ Microbiol 61:1867–1875

    CAS  PubMed  Google Scholar 

  • Volkin DB, Klibanov AM (1989) Mechanism of thermoinactivation of immobilized glucose isomerase. Biotechnol Bioeng 33:1104–1111

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgement

This work was supported by the 21C Frontier Microbial Genomics and Application Center Program, Ministry of Education, Science & Technology, Republic of Korea.

Author information

Authors and Affiliations

  1. Biological Resource Center (BRC), Korean Collection for Type Cultures (KCTC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yuseong-gu, Daejeon, 305-806, South Korea

    Byoung-Chan Kim & Jung Sook Lee

  2. Department of Microbiology and Immunology, Pusan National University School of Medicine, Yangsan, 626-870, Korea

    Sun Nyoung Yu, Kwang Youn Kim & Soon Cheol Ahn

  3. Medical Research Institute, Pusan National University Hospital, Busan, 602-739, South Korea

    Sun Nyoung Yu, Kwang Youn Kim & Soon Cheol Ahn

  4. Department of Biotechnology, Yonsei University, 134 Sinchon-dong, Seodaemun-gu, Seoul, 120-749, South Korea

    Yu-Ryang Pyun

Authors
  1. Byoung-Chan Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sun Nyoung Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kwang Youn Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jung Sook Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yu-Ryang Pyun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Soon Cheol Ahn
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yu-Ryang Pyun or Soon Cheol Ahn.

Electronic supplementary material

Below is the link to the electronic supplementary material.

(DOC 58.5 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, BC., Yu, S.N., Kim, K.Y. et al. Cloning, expression and characterization of xylose isomerase, XylA, from Caldanaerobacter subterraneus subsp. yonseiensis . Biotechnol Lett 32, 929–933 (2010). https://doi.org/10.1007/s10529-010-0255-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-010-0255-3

Keywords

Search

Navigation