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Purification and Characterization of Cellulase from Obligate Halophilic Aspergillus flavus (TISTR 3637) and Its Prospects for Bioethanol Production

  • Amna Bano
  • Xiaoming Chen
  • Sehanat PrasongsukEmail author
  • Ali Akbar
  • Pongtharin Lotrakul
  • Hunsa Punnapayak
  • Muhammad Anwar
  • Sumbal Sajid
  • Imran AliEmail author
Article

Abstract

A cellulase from the extreme obligate halophilic fungus, Aspergillus flavus, isolated from a man-made solar saltern in Phetchaburi, Thailand, was purified by ammonium sulfate precipitation and using Sephadex G-100 gel filtration column chromatography. The cellulase was found to be approximately 55 kDa by SDS-PAGE. Using CMC as a substrate, the specific activity of the cellulase was 62.9 units (U) mg−1 with Vmax and Km values of 37.87 mol min−1 mg−1 and 3.02 mg mL−1, respectively. Characterization of the enzyme revealed it to be an extremozyme, having an optimum activity at pH 10, 60 °C, and 200 g L−1 of NaCl. The enzyme activity was not significantly altered by the addition of divalent metal cations at 2 mM and neither did ß-mercaptoethanol, while EDTA was found strongly inhibiting the cellulase. Compared with commercial cellulase, the purified cellulase from A. flavus was more active in the extremity of conditions, especially at pH 10, 60 °C, and 150 g L−1 NaCl, whereas the commercial cellulase had a very low activity.

Keywords

Halophilic fungi Aspergillus flavus Cellulose Extremozyme 

Notes

Funding Information

This research was funded by the Ratchadapisek Sompoch Endowment Fund (2016), Chulalongkorn University (CU59-049-EN).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Life Sciences and EngineeringSouthwest University of Science and TechnologyMianyangChina
  2. 2.Institute of BiochemistryUniversity of BalochistanQuettaPakistan
  3. 3.Plant Biomass Utilization Research Unit, Botany DepartmentChulalongkorn UniversityBangkokThailand
  4. 4.Department of MicrobiologyUniversity of BalochistanQuettaPakistan
  5. 5.Department of Biology, Faculty of Science and TechnologyUniversitas AirlanggaSurabayaIndonesia

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