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Kinetic Study of Immobilized Cellobiase Produced from Immobilized Wild-Type Trichoderma longibrachiatum

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Abstract

In recent years, there has been an increasing trend toward more efficient utilization of agro-industrial residues, such as sugarcane bagasse, as raw materials for industrial applications. The present study attempts to find out the effect of immobilization of Trichoderma longibrachiatum on enzyme production and immobilization of cellobiase on its kinetic activity. T. longibrachiatum was immobilized on sodium alginate (1, 1.5, and 2 % w/v) beads. The sugarcane bagasse, after autoclaving, was inoculated with 10 ml of immobilized biocatalyst and free cell culture. Maximum cellulase production was observed in immobilized fermentation system after 120 h of incubation. Cellobiase was purified up to 4.8-folds using DEAE-Sepharose column pre-equilibrated in 50 mM Tris–HCl buffer (pH 7.0) and eluted in same buffer by varying the degree of NaCl concentration. The specific activity before and after purification was 0.038 and 0.181 U/mg, respectively. The percent-entrapped activity of calcium alginate immobilized cellobiase was found maximum at 6.5 % (w/v) sodium alginate. The pH optimum of the native free enzyme was 5.5 and entrapped enzyme was 5.0. Immobilized cellobiase exhibited maximum activity at 40 °C at pH 5.5 (42.143 U). The purified enzyme was found to be highly thermostable and was found active over range of 20–80 °C and pH 4.0–8.0. The K m and V max for immobilized cellobiase from T. longibrachiatum were found 44.74 mg/ml and 79.49 mM/ml/min, respectively.

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Authors and Affiliations

  1. Department of Biotechnology, Integral University, Dasauli, Kursi Road, Lucknow, Uttar Pradesh, 226026, India

    Mohammad Ashfaque & Neelam Pathak

  2. Indian Institute of Sugarcane Research, Raibareli Road, P.O. Dilkusha, Lucknow, 226 002, India

    Mohammad Ashfaque & Sushil Solomon

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  1. Mohammad Ashfaque
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  2. Sushil Solomon
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  3. Neelam Pathak
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Correspondence to Mohammad Ashfaque.

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It is to disclose that for any aspect of the submitted work (including but not limited to grants, data monitoring board, study design, manuscript preparation, statistical analysis, etc.) has been financed from a third party (government, commercial, private foundation, etc.). There are no conflicts of interest among the authors.

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Ashfaque, M., Solomon, S. & Pathak, N. Kinetic Study of Immobilized Cellobiase Produced from Immobilized Wild-Type Trichoderma longibrachiatum . Sugar Tech 18, 340–346 (2016). https://doi.org/10.1007/s12355-015-0410-1

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  • DOI: https://doi.org/10.1007/s12355-015-0410-1

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