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Response surface optimization for enhanced production of cellulases with improved functional characteristics by newly isolated Aspergillus niger HN-2

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Abstract

Fungi isolated from partially decayed wood log samples showing characteristic diversity for spore colour, colony morphology and arrangement of spores were assessed for cellulolytic enzyme production. Isolates showing a cellulolytic index of ≥2.0 were assayed for filter paper (FP) cellulase and β-glucosidase (BGL) production. Molecular characterization confirmed the identity of the selected cellulolytic isolate as a strain of Aspergillus niger (A. niger HN-2). Addition of 2 % (w/v) urea enhanced FP and BGL activity by about 20 and 60 %, respectively. Validation studies conducted at parameters (29 °C, pH 5.4, moisture content 72 % and 66 h) optimized through response surface methodology in a solid-state static tray fermentation resulted in FP, BGL, cellobiohydrolase I (CBHI), endoglucanase (EG), xylanase activity and protein content of 25.3 FPU/g ds, 750 IU/g ds, 13.2 IU/g ds, 190 IU/g ds, 2890 IU/g ds and 0.9 mg/ml, respectively. In comparison, A. niger N402 which is a model organism for growth and development studies, produced significantly lower FP, BGL, CBHI, EG, xylanase activity and protein content of 10.0 FPU/g ds, 100 IU/g ds, 2.3 IU/g ds, 50 IU/g ds, 500 IU/g ds and 0.75 mg/ml, respectively under the same process conditions as were used for A. niger HN-2. Process optimization led to nearly 1.8- and 2.2-fold increase in FP and BGL activity, respectively showing promise for cellulase production by A. niger HN-2 at a higher scale of operation. Zymogram analysis revealed two isoforms each for EG and cellobiohydrolase and three isoforms for BGL. Crude cellulase complex produced by A. niger HN-2 exhibited thermostability under acidic conditions showing potential for use in biofuel industry.

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Acknowledgments

Authors thankfully acknowledge the financial assistance received under the NAIP Project (418301-3) funded by World Bank from the Indian Council of Agricultural Research, New Delhi, India for conducting this study.

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Authors and Affiliations

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

    Harinder Singh Oberoi & Rekha Rawat

  2. Department of Microbiology, Guru Nanak Dev University, Amritsar, India

    Bhupinder Singh Chadha

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  1. Harinder Singh Oberoi
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  2. Rekha Rawat
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  3. Bhupinder Singh Chadha
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Correspondence to Harinder Singh Oberoi.

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Oberoi, H.S., Rawat, R. & Chadha, B.S. Response surface optimization for enhanced production of cellulases with improved functional characteristics by newly isolated Aspergillus niger HN-2. Antonie van Leeuwenhoek 105, 119–134 (2014). https://doi.org/10.1007/s10482-013-0060-9

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