Skip to main content
SpringerLink
Log in

Symbiotic Streptomyces sp. TN119 GH 11 xylanase: a new pH-stable, protease- and SDS-resistant xylanase

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A pH-stable and protease-resistant xylanase (XynB119) was identified from Streptomyces sp. TN119, a strain isolated from the gut luminal contents of longhorned beetle (Batocera horsfieldi) larvae. Using the GC TAIL-PCR method, the 1,026-bp coding gene (xynB119) with 67.3% GC content was successfully cloned and expressed in Escherichia coli. It encodes a 341-residue polypeptide with a calculated molecular mass of 35.9 kDa, including a putative 41-residue signal peptide, a catalytic domain of glycosyl hydrolase (GH) family 11, a short Gly/Pro-rich linker, and a family 2 cellulose-binding domain (CBM 2). The deduced amino acid sequence is most similar to (61.9% identity) an endo-1,4-β-xylanase from Streptomyces thermoviolaceus OPC-520. Purified recombinant XynB119 exhibited peak activity at 50°C and pH 7.0, remained stable over a broad pH range (retaining >70% activity after incubation at pH 1.0–11.0 for 1 h at 37°C without substrate), had strong protease resistance (retaining >90% activity after proteolytic treatment at 37°C for 1 h) and SDS resistance (at 100 mM). These properties make XynB119 promising for application in the feed industry and valuable for basic research. Compared to r-XynB119, the r-XynB119 derivative without CBM 2 and linker region (r-XynB119d) exhibited a decreased pH stability of >25% at extreme pHs (pH 1.0–3.0 and pH 11.0–12.0).

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

  1. Thomson JA (1993) Molecular biology of xylan degradation. FEMS Microbiol Rev 104:65–82

    Article  CAS  Google Scholar 

  2. Finn RD, Tate J, Mistry J, Coggill PC, Sammut SJ, Hotz HR, Ceric G, Forslund K, Eddy SR, Sonnhammer ELL, Bateman A (2008) The Pfam protein families database. Nucleic Acids Res 36:D281–D288

    Article  PubMed  CAS  Google Scholar 

  3. Collins T, Gerday C, Feller G (2005) Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 29:3–23

    Article  PubMed  CAS  Google Scholar 

  4. Scriber JM, Slansky F (1981) The nutritional ecology of immature insects. Annu Rev Entomol 26:183–211

    Article  Google Scholar 

  5. Warnecke F, Luginbuhl P, Ivanova N, Ghassemian M, Richardson TH, Stege JT, Cayouette M, McHardy AC, Djordjevic G, Aboushadi N, Sorek R, Tringe SG, Podar M, Martin HG, Kunin V, Dalevi D, Madejska J, Kirton E, Platt D, Szeto E, Salamov A, Barry K, Mikhailova N, Kyrpides NC, Matson EG, Ottesen EA, Zhang XN, Hernandez M, Murillo C, Acosta LG, Rigoutsos I, Tamayo G, Green BD, Chang C, Rubin EM, Mathur EJ, Robertson DE, Hugenholtz P, Leadbetter JR (2007) Metagenomic and functional analysis of hindgut microbiota of a wood-feeding higher termite. Nature 450:560–565

    Article  PubMed  CAS  Google Scholar 

  6. Xu J, Mahowald MA, Ley RE, Lozupone CA, Hamady M, Martens EC, Henrissat B, Coutinho PM, Minx P, Latreille P, Cordum H, Van Brunt A, Kim K, Fulton RS, Fulton LA, Clifton SW, Wilson RK, Knight RD, Gordon JI (2007) Evolution of symbiotic bacteria in the distal human intestine. PLoS Biol 5:1574–1586

    CAS  Google Scholar 

  7. Ferrer M, Golyshina OV, Chernikova TN, Khachane AN, Reyes-Duarte D, Dos Santos V, Strompl C, Elborough K, Jarvis G, Neef A, Yakimov MM, Timmis KN, Golyshin PN (2005) Novel hydrolase diversity retrieved from a metagenome library of bovine rumen microflora. Environ Microbiol 7:1996–2010

    Article  PubMed  CAS  Google Scholar 

  8. Breznak JA, Brune A (1994) Role of microorganisms in the digestion of lignocellulose by termites. Annu Rev Entomol 39:453–487

    Article  CAS  Google Scholar 

  9. Zhou J, Huang H, Meng K, Shi P, Wang Y, Luo H, Yang P, Bai Y, Zhou Z, Yao B (2009) Molecular and biochemical characterization of a novel xylanase from the symbiotic Sphingobacterium sp. TN19. Appl Microbiol Biotechnol 85:323–333

    Article  PubMed  CAS  Google Scholar 

  10. Zhou J, Huang H, Meng K, Shi P, Wang Y, Luo H, Yang P, Bai Y, Yao B (2010) Cloning of a new xylanase gene from Streptomyces sp. TN119 using a modified thermal asymmetric interlaced-PCR specific for GC-rich genes and biochemical characterization. Appl Biochem Biotechnol 160:1277–1292

    Article  PubMed  CAS  Google Scholar 

  11. Heo S, Kwak J, Oh HW, Park DS, Bae KS, Shin DH, Park HY (2006) Characterization of an extracellular xylanase in Paenibacillus sp. HY-8 isolated from an herbivorous longicorn beetle. J Microbiol Biotechnol 16:1753–1759

    CAS  Google Scholar 

  12. Park DS, Oh HW, Heo SY, Jeong WJ, Shin DH, Bae KS, Park HY (2007) Characterization of an extracellular lipase in Burkholderia sp. HY-10 isolated from a longicorn beetle. J Microbiol 45:409–417

    PubMed  CAS  Google Scholar 

  13. Zhou J, Meng K, Yang P, Shi P, Wang Y, Luo H, Yao B (2010) Characterization of a chromosomal segment showing xylanolytic activity from the symbiotic Sphingobacterium sp. TN19. World J Microbiol Biotechnol 26:761–765

    Article  CAS  Google Scholar 

  14. Bendtsen JD, Nielsen H, von Heijne G, Brunak S (2004) Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340:783–795

    Article  PubMed  Google Scholar 

  15. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    PubMed  CAS  Google Scholar 

  16. Finn RD, Mistry J, Tate J, Coggill P, Heger A, Pollington JE, Gavin OL, Gunasekaran P, Ceric G, Forslund K, Holm L, Sonnhammer EL, Eddy SR, Bateman A (2010) The Pfam protein families database. Nucleic Acids Res 38:D211–D222

    Article  PubMed  CAS  Google Scholar 

  17. Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948

    Article  PubMed  CAS  Google Scholar 

  18. Guex N, Peitsch MC (1997) SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 18:2714–2723

    Google Scholar 

  19. Eisenberg D, Luthy R, Bowie JU (1997) VERIFY3D: assessment of protein models with three-dimensional profiles. Methods Enzymol 277:396–404

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  23. McCarthy AA, Morris DD, Bergquist PL, Baker EN (2000) Structure of XynB, a highly thermostable β-1, 4-xylanase from Dictyoglomus thermophilum Rt46B.1, at 1.8 angstrom resolution. Acta Crystallogr Sect D Biol Crystallogr 56:1367–1375

    Article  CAS  Google Scholar 

  24. Bai YG, Wang JS, Zhang ZF, Yang PL, Shi PJ, Luo HY, Meng K, Huang HQ, Yao B (2010) A new xylanase from thermoacidophilic Alicyclobacillus sp. A4 with broad-range pH activity and pH stability. J Ind Microbiol Biotechnol 37:187–194

    Article  PubMed  CAS  Google Scholar 

  25. Blanco J, Coque JJR, Velasco J, Martin JF (1997) Cloning, expression in Streptomyces lividans and biochemical characterization of a thermostable endo-β-1, 4-xylanase of Thermomonospora alba ULJB1 with cellulose-binding ability. Appl Microbiol Biotechnol 48:208–217

    Article  PubMed  CAS  Google Scholar 

  26. Ito K, Ogasawara H, Sugimoto T, Ishikawa T (1992) Purification and properties of acid stable xylanases from Aspergillus Kawachii. Biosci Biotechnol Biochem 56:547–550

    Article  CAS  Google Scholar 

  27. Jiang ZQ, Kobayashi A, Ahsan MM, Lite L, Kitaoka M, Hayashi K (2001) Characterization of a thermostable family 10 endo-xylanase (XynB) from Thermotoga maritima that cleaves p-nitrophenyl-β-d-xyloside. J Biosci Bioeng 92:423–428

    Article  CAS  Google Scholar 

  28. Li N, Shi PJ, Yang PL, Wang YR, Luo HY, Bai YG, Zhou ZG, Yao B (2009) A xylanase with high pH stability from Streptomyces sp. S27 and its carbohydrate-binding module with/without linker-region-truncated versions. Appl Microbiol Biotechnol 83:99–107

    Article  PubMed  CAS  Google Scholar 

  29. Li N, Yang PL, Wang Y, Luo HY, Meng K, Wu NF, Fan YL, Yao B (2008) Cloning, expression, and characterization of protease-resistant xylanase from Streptomyces fradiae var. k11. J Microbiol Biotechnol 18:410–416

    PubMed  CAS  Google Scholar 

  30. Meng X, Shao ZZ, Hong YZ, Lin L, Li CJ, Liu ZD (2009) A novel pH-stable, bifunctional xylanase isolated from a deep-sea microorganism, Demequina sp. JK4. J Microbiol Biotechnol 19:1077–1084

    PubMed  CAS  Google Scholar 

  31. Kittur FS, Mangala SL, Abu Rus’d A, Kitaoka M, Tsujibo H, Hayashi K (2003) Fusion of family 2b carbohydrate-binding module increases the catalytic activity of a xylanase from Thermotoga maritima to soluble xylan. FEBS Lett 549:147–151

    Article  PubMed  CAS  Google Scholar 

  32. Morgavi DP, Beauchemin KA, Nsereko VL, Rode LM, McAllister TA, Iwaasa AD, Wang Y, Yang WZ (2001) Resistance of feed enzymes to proteolytic inactivation by rumen microorganisms and gastrointestinal proteases. J Anim Sci 79:1621–1630

    PubMed  CAS  Google Scholar 

  33. Luo HY, Wang Y, Li J, Wang H, Yang J, Yang YH, Huang HQ, Fan YL, Yao B (2009) Cloning, expression and characterization of a novel acidic xylanase, XYL11B, from the acidophilic fungus Bispora sp. MEY-1. Enzyme Microb Technol 45:126–133

    Article  CAS  Google Scholar 

  34. Manning M, Colon W (2004) Structural basis of protein kinetic stability: resistance to sodium dodecyl sulfate suggests a central role for rigidity and a bias toward β-sheet structure. Biochemistry 43:11248–11254

    Article  PubMed  CAS  Google Scholar 

  35. Jaswal SS, Sohl JL, Davis JH, Agard DA (2002) Energetic landscape of α-lytic protease optimizes longevity through kinetic stability. Nature 415:343–346

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the Earmarked Fund for Modern Agro-industry Technology Research System (NYCYTX-42-G2-05), Key Program of Transgenic Plant Breeding (2009ZX08003-020B), and the National High Technology Research and Development Program of China (863 Program; grant 2007AA100601).

Author information

Author notes

  1. Junpei Zhou

    Present address: College of Life Sciences, Yunnan Normal University, No. 298 December 1st Street, Kunming, 650092, People’s Republic of China

Authors and Affiliations

  1. Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081, People’s Republic of China

    Junpei Zhou, Pengjun Shi, Rui Zhang, Huoqing Huang, Kun Meng, Peilong Yang & Bin Yao

Authors
  1. Junpei Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pengjun Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rui Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Huoqing Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kun Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Peilong Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bin Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bin Yao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhou, J., Shi, P., Zhang, R. et al. Symbiotic Streptomyces sp. TN119 GH 11 xylanase: a new pH-stable, protease- and SDS-resistant xylanase. J Ind Microbiol Biotechnol 38, 523–530 (2011). https://doi.org/10.1007/s10295-010-0795-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-010-0795-5

Keywords

Search

Navigation