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Applied Biochemistry and Biotechnology

, Volume 187, Issue 1, pp 239–252 | Cite as

Expression and Extracellular Secretion of Endo-glucanase and Xylanase by Zymomonas mobilis

  • Tatsaporn TodhanakasemEmail author
  • Apinya Sowatad
  • Pattanop Kanokratana
  • Phattara-orn Havanapan
  • Verawat Champreda
Article
  • 198 Downloads

Abstract

Recombinant Zymomonas mobilis (pGEX-4T-3 BI 120-2) was constructed to encode endo-glucanase (CelA) and endo-xylanase (Xyn11) from Z. mobilis ZM4 (ATCC 31821) and an uncultured bacterium. The recombinant was genetically engineered with the N-terminus of a predicted SecB-dependent (type II) secretion signal from phoC of Z. mobilis to translocate the enzymes extracellularly. Both the enzymes were characterized regarding their functional optimum pH and temperature, with the highest multi-enzyme activities at pH 6.0 and a temperature of 30 °C, which approximates the optimum conditions for ethanol production by Z. mobilis. The crude intracellular and extracellular fractions of the recombinant were characterized in terms of substrate specificity using carboxymethyl cellulose (CMC), beechwood xylan, filter paper, Avicel, and pretreated rice straw. The crude extracellular and intracellular enzymes with cellulolytic and xylanolytic activities were more robustly produced and secreted from the recombinant strain compared to the wild-type and ampicillin-sensitive strains, using CMC and beechwood xylan as the substrates. Ethanol production by the recombinant strain was greater than the production by the wild-type strain when pretreated rice straw was used as a sole carbon source.

Keywords

Zymomonas mobilis Cellulase Xylanase Extracellular Ethanol production 

Notes

Acknowledgments

This work was financially supported by an Assumption University Research Grant (RP 59-002), Thailand.

Funding

The Z. mobilis ZM4 (NRRL B-14023) used in this study was obtained from Professor Pornthap Thanonkeo, Khon Kaen University, Thailand.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Agro-Industry, Faculty of BiotechnologyAssumption UniversityBangkokThailand
  2. 2.National Center for Genetic Engineering and Biotechnology (BIOTEC)Klong LuangThailand
  3. 3.Institute of Molecular BiosciencesMahidol UniversitySalayaThailand

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