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

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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.

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Funding

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

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Author notes

  1. Amna Bano and Xiaoming Chen authors are considered as first author, having equal contribution as principal authors.

Authors and Affiliations

  1. School of Life Sciences and Engineering, Southwest University of Science and Technology, Mianyang, 621010, Sichuan, China

    Amna Bano, Xiaoming Chen & Imran Ali

  2. Institute of Biochemistry, University of Balochistan, Quetta, 87300, Pakistan

    Amna Bano, Muhammad Anwar, Sumbal Sajid & Imran Ali

  3. Plant Biomass Utilization Research Unit, Botany Department, Chulalongkorn University, Bangkok, 10330, Thailand

    Sehanat Prasongsuk, Pongtharin Lotrakul, Hunsa Punnapayak & Imran Ali

  4. Department of Microbiology, University of Balochistan, Quetta, 87300, Pakistan

    Ali Akbar

  5. Department of Biology, Faculty of Science and Technology, Universitas Airlangga, Surabaya, 60115, Indonesia

    Hunsa Punnapayak

Authors
  1. Amna Bano
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  2. Xiaoming Chen
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  3. Sehanat Prasongsuk
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  4. Ali Akbar
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  5. Pongtharin Lotrakul
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  6. Hunsa Punnapayak
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  7. Muhammad Anwar
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  8. Sumbal Sajid
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Correspondence to Sehanat Prasongsuk or Imran Ali.

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Bano, A., Chen, X., Prasongsuk, S. et al. Purification and Characterization of Cellulase from Obligate Halophilic Aspergillus flavus (TISTR 3637) and Its Prospects for Bioethanol Production. Appl Biochem Biotechnol 189, 1327–1337 (2019). https://doi.org/10.1007/s12010-019-03086-y

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  • DOI: https://doi.org/10.1007/s12010-019-03086-y

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