Advertisement

Characterization of a novel multi-domain xylanase from Clostridium clariflavum with application in hydrolysis of corn cobs

  • Yulu Liu
  • Yawu Sun
  • Huaguang Wang
  • Lei TangEmail author
Original Research Paper
  • 9 Downloads

Abstract

Objectives

To develop a novel multi-catalytic domain (CD) xylanase Xyn2083 from Clostridium clariflavum by expression of its truncated forms in Escherichia coli and cooperation of xylanase with cellulase in the hydrolysis of waste lignocellulosic resources.

Results

Xyn2083 has two glycoside hydrolase family (GH) domains GH11 and GH10. These two catalytic domains functioned synergistically in xylan hydrolysis. The recombinant protein with GH11 domain, Xyn2083GH11, had the highest xylanase activity among three constructed truncated forms. The deletion of N-terminal extra amino acid residues of Xyn2083GH11 decreased catalytic activity as well as the stability of the enzyme. The hydrolysis rates of cellulose and xylan in the pretreated corn cobs were 90.56% and 72.80% with the addition of Xyn2083GH11 and cellulase, whereas those were 67.95% and 34.45% using sole cellulase respectively. The structural analysis of substrates indicated that the addition of Xyn2083GH11 led to a looser structure and more exposure of crystal cellulose for cellulase to approach.

Conclusions

Since the native multi-CDs’ xylanases are rare, the thermostable Xyn2083 provides a good source for functional studies of two CDs coexisted in one xylanase and for potential applications after modification.

Keywords

Clostridium clariflavum Corn cobs Xylan Xylanase 

Notes

Acknowledgements

This work was supported by the Program of Introducing Talents of Discipline to Universities (111-2-06), the National First-class Discipline Program of Light Industry Technology and Engineering (LITE2018-27), and Top-notch Academic Programs Project of Jiangsu Higher Education Institutions (PPZY2015A056).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10529_2019_2721_MOESM1_ESM.docx (40 kb)
Supplementary Table 1—Primers used for amplification of the genes of Xyn2083 derivatives. Supplementary Fig. 1—Xyn2083GH11and its truncated forms. Supplementary material 1 (DOCX 40 kb)

References

  1. Biely P, Vršanská M, Tenkanen M, Kluepfel D (1997) Endo-β-1, 4-xylanase families: differences in catalytic properties. J Biotechnol 57:151–166CrossRefGoogle Scholar
  2. Chang S, Guo Y, Wu B, He B (2017) Extracellular expression of alkali tolerant xylanase from Bacillus subtilis Lucky9 in E. coli and application for xylo oligosaccharides production from agro-industrial waste. Int J Biol Macromol 96:249–256CrossRefGoogle Scholar
  3. Feng H, Sun Y, Zhi Y, Mao L, Luo Y, Xu L, Wang L, Zhou P (2015) Expression and characterization of a novel endo-1,4-β-xylanase produced by Streptomyces griseorubens JSD-1 isolated from compost-treated soil. Ann Microbiol 65:1771–1779CrossRefGoogle Scholar
  4. Geng A, Wang H, Wu J, Xie R, Sun J (2017) Characterization of a β-xylosidase from Clostridium clariflavum and its application in xylan hydrolysis. BioResources 12:9253–9262Google Scholar
  5. Goncalves GA, Takasugi Y, Jia L, Noda S, Tanaka T, Ichinose H, Kamiya N (2015) Synergistic effect and application of xylanases as accessory enzymes to enhance the hydrolysis of pretreated bagasse. Enzyme Microb Technol 72:16–24CrossRefGoogle Scholar
  6. Hu J, Arantes V, Pribowo A, Saddler JN (2013) The synergistic action of accessory enzymes enhances the hydrolytic potential of a “cellulase mixture” but is highly substrate specific. Biotechnol Biofuels 6:112CrossRefGoogle Scholar
  7. Izquierdo JA, Goodwin L, Davenport KW et al (2012) Complete genome sequence of Clostridium clariflavum DSM 19732. Stand Genomic Sci 6:104–115CrossRefGoogle Scholar
  8. Juturu V, Wu JC (2012) Microbial xylanases: engineering, production and industrial applications. Biotechnol Adv 30:1219–1227CrossRefGoogle Scholar
  9. Maitan-Alfenas GP, Visser EM, Guimarães VM (2015) Enzymatic hydrolysis of lignocellulosic biomass: converting food waste in valuable products. Curr Opin Food Sci 1:44–49CrossRefGoogle Scholar
  10. Melo-Silveira RF, Fidelis GP, Viana RLS, Soeiro VC, da Silva RG, Machado D, Costa LS, Ferreira CV, Rocha HAO (2014) Antioxidant and antiproliferative activities of methanolic extract from a neglected agricultural products: corn cobs. Molecules 19:5360–5378CrossRefGoogle Scholar
  11. Shiratori H, Sasaya K, Ohiwa H, Ayame S, Miya A, Beppu T, Ueda K (2009) Clostridium clariflavum sp. nov. and Clostridium caenicola sp. nov., moderately thermophilic, cellulose-/cellobiose-digesting bacteria isolated from methanogenic sludge. Int J Syst Evol Microbiol 59:1764–1770CrossRefGoogle Scholar
  12. Wang G, Ren Y, Ng TB, Streit WR, Ye X (2019) High-throughput amplicon sequencing demonstrates extensive diversity of xylanase genes in the sediment of soda lake Dabusu. Biotechnol Lett 41:409–418CrossRefGoogle Scholar
  13. Zhang Q, He J, Tian M, Mao Z, Tang L, Zhang J, Zhang H (2011) Enhancement of methane production from cassava residues by biological pretreatment using a constructed microbial consortium. Bioresour Technol 102:8899–8906CrossRefGoogle Scholar
  14. Zhang Q, Li H, Zhu X, Lai F, Zhai Z, Wang Y (2016) Exploration of the key functional proteins from an efficient cellulolytic microbial consortium using dilution-to-extinction approach. J Environ Sci (China) 43:199–207CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Key Laboratory of Industrial Biotechnology, Ministry of EducationJiangnan UniversityWuxiChina
  2. 2.School of BiotechnologyJiangnan UniversityWuxiChina

Personalised recommendations