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Endoglucanase Produced by Bacillus subtilis Strain CBS31: Biochemical Characterization, Thermodynamic Study, Enzymatic Hydrolysis, and Bio-industrial Applications

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  • Enzyme Technology and Protein Engineering
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Abstract

Microbial cellulases have become the mainstream biocatalysts due to their complex nature and widespread industrial applications. Here, homogeneous endoglucanase GluCB31 from Bacillus subtilis subsp. inaquosorum CBS31 was studied. GluCB31 was purified to 17.68-fold with an 8.33% yield and a specific activity of 1066.37 U/mg. Biochemical properties of GluCB31 were performed and the results are as follows; molecular mass of 35 kDa with an optimum pH at 7.5 and temperature at 50°C. GluCB31 was immobilized in calcium alginate gel and it exhibited the highest activity at 10°C higher temperature than soluble enzyme, as the entrapment in alginate gel made GluCB31 more stable. Kinetic studies showed the Vmax of 1293.33 ± 2.51 U/mg and Km of 0.0183 mg/mL. Enzymatic activity was activated by Tween-20 (106.7%), Tween-80 (111.6%), Triton X-100 (142.3%), SDS (135.5%), Mg++ (185.7%), Cu++ (167.6%), Zn++ (153.7%), Mn++ (106.3%), Ba++ (181.9%), Ni++ (107.2%) while inhibited by Fe++ (15.8%), β-mercaptoethanol (46.8%), EDTA (54.5%). Enthalpy, free energy, and entropy of activation were calculated to be 38.526 kJmol-1, 44.187 kJmol-1, and -17.518 Jmol-1K-1 respectively. Also, ΔGE-S and ΔGE-T were found to be -10.75 kJmol-1 and -45.92 kJmol-1 respectively. A low ΔS, ΔGE-S, and ΔGE-T values were signified enzyme-catalyzed reaction occurs at a fast rate and the existence of the enzyme in its stable state. Cellobiose was the major end product of hydrolysis. These attributes of GluCB31 demonstrated the diversity of catalytic activities and serve in various biotechnological processes, thus deserve to be developed as a bio-industrial agent.

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Acknowledgements

This work was carried out with the support of “Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ01319101 and PJ01331001)” Rural Development Administration, Republic of Korea and National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (NRF-2018R1D1A1B05050137).

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  1. Authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacy, College of Pharmacy, Chosun University, Gwangju, 61452, Korea

    Sudip Regmi, Yoon Seok Choi, Young Kyun Kim, Md Maruf Khan, Sang Hun Lee & Jin Cheol Yoo

  2. Department of Pharmacy, Mokpo National University, Muan, 58554, Korea

    Seung Sik Cho

  3. Center for Nutraceutical and Pharmaceutical Materials, Myongji University, Yongin, 17058, Korea

    Ying-Yu Jin & Joo-Won Suh

  4. Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong, 27709, Korea

    Dae Young Lee

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Regmi, S., Choi, Y.S., Kim, Y.K. et al. Endoglucanase Produced by Bacillus subtilis Strain CBS31: Biochemical Characterization, Thermodynamic Study, Enzymatic Hydrolysis, and Bio-industrial Applications. Biotechnol Bioproc E 25, 104–116 (2020). https://doi.org/10.1007/s12257-019-0338-5

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