Journal of Industrial Microbiology & Biotechnology

, Volume 40, Issue 10, pp 1083–1093 | Cite as

A novel neutral xylanase with high SDS resistance from Volvariella volvacea: characterization and its synergistic hydrolysis of wheat bran with acetyl xylan esterase

  • Fei Zheng
  • Jingxuan Huang
  • Yuhao Yin
  • Shaojun Ding


A neutral xylanase (XynII) from Volvariella volvacea was identified and characterized. Unlike other modular xylanases, it consists of only a single GH10 catalytic domain with a unique C-terminal sequence (W-R-W-F) and a phenylalanine and proline-rich motif (T-P-F-P-P-F) at N-terminus, indicating that it is a novel GH10 xylanase. XynII exhibited optimal activity at pH 7 and 60 °C and stability over a broad range of pH 4.0–10.0. XynII displayed extreme highly SDS resistance retaining 101.98, 92.99, and 69.84 % activity at the presence of 300 mM SDS on birchwood, soluble oat spelt, and beechwood xylan, respectively. It remained largely intact after 24 h of incubation with proteinase K at a protease to protein ratio of 1:50 at 37 °C. The kinetic constants Km value towards beechwood xylan was 0.548 mg ml−1, and the kcat/Km ratio, reflecting the catalytic efficiency of the enzyme, was 126.42 ml mg−1 s−1 at 60 °C. XynII was a true endo-acting xylanase lacking cellulase activity. It has weak activity towards xylotriose but efficiently hydrolyzed xylans and xylooligosaccharides larger than xylotriose mainly to xylobiose. Synergistic action with acetyl xylan esterase (AXEI) from V. volvacea was observed for de-starched wheat bran. The highest degree of synergy (DS 1.42) was obtained in sequential reactions with AXEI digestion preceding XynII. The high SDS resistance and intrinsic stability suggested XynII may have potential applications in various industrial processes especially for the detergent and textile industries and animal feed industries.


Volvariella volvacea Xylanase SDS resistance Synergistic action Acetyl xylan esterase 



Glycoside hydrolase family


























Expressed sequence tag


Sodium dodecyl sulfate


Sodium dodecyl sulfate-polyacrylamide gel electrophoresis


High-performance anion exchange chromatography with pulsed amperometric detection



This work was supported by a research grant (no. 31270628) from the National Natural Science Foundation of China, a Project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, and the Doctorate Fellowship Foundation of Nanjing Forestry University.


  1. 1.
    Bai Y, Wang J, Zhang Z, Yang P, Shi P, Luo H, Meng K, Huang H, Yao B (2010) A new xylanase from thermoacidophilic Alicyclobacillus sp. A4 with broad-range pH activity and pH stability. J Ind Microbiol Biotechnol 37(2):187–194PubMedCrossRefGoogle Scholar
  2. 2.
    Bhardwaj A, Leelavathi S, Mazumdar-Leighton S, Ghosh A, Ramakumar S, Reddy VS (2010) The critical role of N- and C-terminal contact in protein stability and folding of a family 10 xylanase under extreme conditions. PLoS One 5(6):e11347PubMedCrossRefGoogle Scholar
  3. 3.
    Biely P (1985) Microbial xylanolytic systems. Trends Biotechnol 3(11):286–290CrossRefGoogle Scholar
  4. 4.
    Chao-Hsun Y, Wen-Hsiung L (2008) Purification and properties of an acetylxylan esterase from Thermobifida fusca. Enzyme Microb Technols 42(2):181–186CrossRefGoogle Scholar
  5. 5.
    Chapla D, Pandit P, Shah A (2012) Production of xylooligosaccharides from corncob xylan by fungal xylanase and their utilization by probiotics. Bioresour Technol 115:215–221PubMedCrossRefGoogle Scholar
  6. 6.
    Chi W-J, Park D, Chang Y-K, Hong S-K (2012) A novel alkaliphilic xylanase from the newly isolated mesophilic Bacillus sp. MX47: production, purification, and characterization. Appl Biochem Biotechnol 168(4):899–909PubMedCrossRefGoogle Scholar
  7. 7.
    Collins T, Gerday C, Feller G (2005) Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 29(1):3–23PubMedCrossRefGoogle Scholar
  8. 8.
    Coughlan MP, Hazlewood GP (1993) Beta-1,4-D-xylan-degrading enzyme systems: biochemistry, molecular biology and applications. Biotechnol Appl Biochem 17(3):259–289PubMedGoogle Scholar
  9. 9.
    Cybinski DH, Layton I, Lowry JB, Dalrymple BP (1999) An acetylxylan esterase and a xylanase expressed from genes cloned from the ruminal fungus Neocallimastix patriciarum act synergistically to degrade acetylated xylans. Appl Microbiol Biotechnol 52(2):221–225PubMedCrossRefGoogle Scholar
  10. 10.
    Ding S, Cao J, Zhou R, Zheng F (2007) Molecular cloning, and characterization of a modular acetyl xylan esterase from the edible straw mushroom Volvariella volvacea. FEMS Microbiol Lett 274(2):304–310PubMedCrossRefGoogle Scholar
  11. 11.
    Dupont C, Daigneault N, Shareck F, Morosoli R, Kluepfel D (1996) Purification and characterization of an acetyl xylan esterase produced by Streptomyces lividans. Biochem J 319(3):881–886PubMedGoogle Scholar
  12. 12.
    Feia A, Novroski N (2013) The evaluation of possible false positives with detergents when performing amylase serological testing on clothing. J Forensic Sci 58:S183–S185PubMedCrossRefGoogle Scholar
  13. 13.
    Fujimoto Z, Kuno A, Kaneko S, Yoshida S, Kobayashi H, Kusakabe I, Mizuno H (2000) Crystal structure of Streptomyces olivaceoviridis E-86 β-xylanase containing xylan-binding domain. J Mol Biol 300(3):575–585PubMedCrossRefGoogle Scholar
  14. 14.
    Heck JX, Flôres SH, Hertz PF, Ayub MAZ (2005) Optimization of cellulase-free xylanase activity produced by Bacillus coagulans BL69 in solid-state cultivation. Process Biochem 40(1):107–112CrossRefGoogle Scholar
  15. 15.
    Jennings PA, Wright PE (1993) Formation of a molten globule intermediate early in the kinetic folding pathway of apomyoglobin. Science 262(5135):892–896PubMedCrossRefGoogle Scholar
  16. 16.
    Joseph B, Ramteke PW, Thomas G (2008) Cold active microbial lipases: some hot issues and recent developments. Biotechnol Adv 26(5):457–470PubMedCrossRefGoogle Scholar
  17. 17.
    Juturu V, Wu JC (2012) Microbial xylanases: engineering, production and industrial applications. Biotechnol Adv 30(6):1219–1227PubMedCrossRefGoogle Scholar
  18. 18.
    Kamal Kumar B, Balakrishnan H, Rele MV (2004) Compatibility of alkaline xylanases from an alkaliphilic Bacillus NCL (87-6-10) with commercial detergents and proteases. J Ind Microbiol Biotechnol 31(2):83–87PubMedCrossRefGoogle Scholar
  19. 19.
    Kaneko S, Ichinose H, Fujimoto Z, Kuno A, Yura K, Go M, Mizuno H, Kusakabe I, Kobayashi H (2004) Structure and function of a family 10 β-xylanase chimera of Streptomyces olivaceoviridis E-86 FXYN and Cellulomonas fimi Cex. J Biol Chem 279(25):26619–26626PubMedCrossRefGoogle Scholar
  20. 20.
    Kasana RC (2010) Proteases from psychrotrophs: an overview. Crit Rev Microbiol 36(2):134–145PubMedCrossRefGoogle Scholar
  21. 21.
    Krishna MMG, Englander SW (2005) The N-terminal to C-terminal motif in protein folding and function. Proc Natl Acad Sci USA 102(4):1053–1058PubMedCrossRefGoogle Scholar
  22. 22.
    Lamsal B, Madl R, Tsakpunidis K (2011) Comparison of feedstock pretreatment performance and its effect on soluble sugar availability. Bioenerg Res 4(3):193–200CrossRefGoogle Scholar
  23. 23.
    Liu C-L, Shen C-R, Hsu F-F, Chen J-K, Wu P-T, Guo S-H, Lee W-C, Yu F-W, Mackey ZB, Turk J, Gross ML (2009) Isolation and identification of two novel SDS-resistant secreted chitinases from Aeromonas schubertii. Biotechnol Prog 25(1):124–131PubMedCrossRefGoogle Scholar
  24. 24.
    Manning M, Colón 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(35):11248–11254PubMedCrossRefGoogle Scholar
  25. 25.
    Moraïs S, Barak Y, Caspi J, Hadar Y, Lamed R, Shoham Y, Wilson DB, Bayer EA (2010) Contribution of a xylan-binding module to the degradation of a complex cellulosic substrate by designer cellulosomes. Appl Environ Microbiol 76(12):3787–3796PubMedCrossRefGoogle Scholar
  26. 26.
    Rättö M, Poutanen K (1988) Production of mannan-degrading enzymes. Biotechnol Lett 10(9):661–664CrossRefGoogle Scholar
  27. 27.
    Ratanakhanokchai K, Kyu KL, Tanticharoen M (1999) Purification and properties of a xylan-binding endoxylanase from alkaliphilic Bacillus sp. Strain K-1. Appl Environ Microbiol 65(2):694–697PubMedGoogle Scholar
  28. 28.
    Raweesri P, Riangrungrojana P, Pinphanichakarn P (2008) Alpha-L-Arabinofuranosidase from Streptomyces sp. PC22: Purification, characterization and its synergistic action with xylanolytic enzymes in the degradation of xylan and agricultural residues. Bioresour Technol 99(18):8981–8986PubMedCrossRefGoogle Scholar
  29. 29.
    Sørensen HR, Pedersen S, Meyer AS (2007) Synergistic enzyme mechanisms and effects of sequential enzyme additions on degradation of water insoluble wheat arabinoxylan. Enzyme Microb Technol 40(4):908–918CrossRefGoogle Scholar
  30. 30.
    Sandrim VC, Rizzatti ACS, Terenzi HF, Jorge JA, Milagres AMF, Polizeli MLTM (2005) Purification and biochemical characterization of two xylanases produced by Aspergillus caespitosus and their potential for kraft pulp bleaching. Process Biochem 40(5):1823–1828CrossRefGoogle Scholar
  31. 31.
    Shallom D, Shoham Y (2003) Microbial hemicellulases. Curr Opin Microbiol 6(3):219–228PubMedCrossRefGoogle Scholar
  32. 32.
    Song Y, Lee YG, Choi IS, Lee KH, Cho EJ, Bae H-J (2013) Heterologous expression of endo-1,4-β-xylanase A from Schizophyllum commune in Pichia pastoris and functional characterization of the recombinant enzyme. Enzyme Microb Technol 52(3):170–176PubMedCrossRefGoogle Scholar
  33. 33.
    St John FJ, González JM, Pozharski E (2010) Consolidation of glycosyl hydrolase family 30: a dual domain 4/7 hydrolase family consisting of two structurally distinct groups. FEBS Lett 584(21):4435–4441PubMedCrossRefGoogle Scholar
  34. 34.
    Viikari L, Ranua M, Kantelinen A, Sundquist J, Linko M (1986) Bleaching with enzymes. In: Paper presented at the Proceedings of the 3rd international conference biotechnology in the pulp and paper industry, STFI, Stockholm, 1986Google Scholar
  35. 35.
    Wang S-Y, Hu W, Lin X-Y, Wu Z-H, Li Y-Z (2012) A novel cold-active xylanase from the cellulolytic myxobacterium Sorangium cellulosum So9733-1: gene cloning, expression, and enzymatic characterization. Appl Microbiol Biotechnol 93(4):1503–1512PubMedCrossRefGoogle Scholar
  36. 36.
    Zhang J, Siika-aho M, Tenkanen M, Viikari L (2011) The role of acetyl xylan esterase in the solubilization of xylan and enzymatic hydrolysis of wheat straw and giant reed. Biotechnol Biofuels 4(1):60PubMedCrossRefGoogle Scholar
  37. 37.
    Zheng F, Ding S (2013) Processivity and enzymatic mode of a glycoside hydrolase family 5 endoglucanase from Volvariella volvacea. Appl Environ Microbiol 79(3):989–996PubMedCrossRefGoogle Scholar
  38. 38.
    Zhou J, Gao Y, Dong Y, Tang X, Li J, Xu B, Mu Y, Wu Q, Huang Z (2012) A novel xylanase with tolerance to ethanol, salt, protease, SDS, heat, and alkali from actinomycete Lechevalieria sp. HJ3. J Ind Microbiol Biotechnol 39(7):965–975PubMedCrossRefGoogle Scholar
  39. 39.
    Zhou J, Shi P, Zhang R, Huang H, Meng K, Yang P, Yao B (2011) Symbiotic Streptomyces sp. TN119 GH 11 xylanase: a new pH-stable, protease- and SDS-resistant xylanase. J Ind Microbiol Biotechnol 38(4):523–530PubMedCrossRefGoogle Scholar

Copyright information

© Society for Industrial Microbiology and Biotechnology 2013

Authors and Affiliations

  • Fei Zheng
    • 1
  • Jingxuan Huang
    • 1
  • Yuhao Yin
    • 1
  • Shaojun Ding
    • 1
  1. 1.Department of Biological EngineeringNanjing Forestry UniversityNanjingChina

Personalised recommendations