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An acidothermophilic functionally active novel GH12 family endoglucanase from Aspergillus niger HO: purification, characterization and molecular interaction studies

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Abstract

Endoglucanase (EG) from Aspergillus niger HO was sequentially purified through ultrafiltration, ion exchange and size exclusion chromatography to homogeneity, with an overall recovery of 18 %. The purified EG was a monomeric protein with a molecular weight of about 55 kDa. The enzyme was optimally active at pH 3.5 and 70 °C with a half life (t1/2) of 3 h and Km value of 2.5 mg/ml. Metal ions, such as Ca2+ and Co2+ helped in enzyme induction, while Hg2+ and Cu2+ strongly inhibited the enzyme activity. Peptide mass fingerprinting results revealed that the purified EG is a novel enzyme that belongs to family 12 of glycoside hydrolase (GH12). Molecular docking studies indicated the presence of Glu116 and Glu204 as important determinant residues for the functional interaction with carboxymethylcellulose and showed hydrogen bonding with Asp99, Glu116, Glu204 and hydrophobic interactions with Trp22, Val58, Tyr61, Phe101, Met118, Trp120, Pro129, Ile130, Thr160 and Phe206. Hydrolysis of 2 % CMC with purified acidothermophilic EG at its optimum temperature and pH resulted in complete hydrolysis within 2 h yielding 18 % cellotriose, 72 % cellobiose and 10 % glucose as evident from HPLC analysis. In comparison to most of the EGs reported in literature, EG from A. niger HO exhibited higher thermostability. The acidothermophilic nature of this enzyme makes it potentially useful for industrial applications.

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Acknowledgments

Authors thankfully acknowledge the financial assistance received from the NAIP sub-project (4183) funded by World Bank through Indian Council of Agricultural Research, Government of India for conducting this study.

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  1. Sunil Kumar

    Present address: National Bureau of Agriculturally Important Microorganisms, Mau Nath Bhanjan, 275101, Uttar Pradesh, India

Authors and Affiliations

  1. Central Institute of Post Harvest Engineering and Technology, P.O. PAU, Ludhiana, 141 004, Punjab, India

    Rekha Rawat & Harinder Singh Oberoi

  2. Institute of Life Sciences, Bhubaneswar, India

    Sunil Kumar

  3. Department of Microbiology, Guru Nanak Dev University (GNDU), Amritsar, 143 005, Punjab, India

    Bhupinder Singh Chadha

  4. Centre for Agricultural Bioinformatics, Indian Agricultural Statistical Research Institute, New Delhi, 110 012, India

    Dinesh Kumar

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  1. Rekha Rawat
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Correspondence to Harinder Singh Oberoi.

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Rawat, R., Kumar, S., Chadha, B.S. et al. An acidothermophilic functionally active novel GH12 family endoglucanase from Aspergillus niger HO: purification, characterization and molecular interaction studies. Antonie van Leeuwenhoek 107, 103–117 (2015). https://doi.org/10.1007/s10482-014-0308-z

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