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Mutagenesis and Immobilization of ChitB-Protease for Induced De-staining and Goat Skin Dehairing Potentialities

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Abstract

Proteases with alkaline nature and requisite catalytic characteristics have been the supreme interest of numerous industrial sectors. The present study was focused on justifying the following aims, i.e., (1) protease producing strain improvement via physical and chemical mutagenesis, (2) protease optimization, purification, and immobilization on chitosan beads, and (3) de-staining and goat skin dehairing potentialities of chitosan beads immobilized protease (ChitB-protease). Briefly, a protease produced by a UV mutant Bacillus sp. MS1 was purified and characterized to present its scope for detergent and leather sectors. Two-step purification resulted up to 4.1-fold purification and 41% yield of Bacillus sp. MS1 protease. The purified fraction was identified as a monomeric with an apparent molecular weight of 28 kDa on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The protease was entrapped in glutaraldehyde-activated chitosan beads to improve its stability and reusability. The ChitB-protease represented the highest activity and immobilization efficiency of 10.31 U/mL and 86.43 ± 1.15%, respectively. ChitB-protease exhibited better catalytic activity in a broader pH range and higher temperature than free enzyme. Moreover, a significant improvement in the thermal resistance was also recorded after immobilization. ChitB-protease retained 70.53% of its initial activity after seven washings. SEM morphological analysis further confirmed the recycling efficiency of ChitB-protease. Furthermore, a complete de-staining and dehairing were observed within the short incubation period. These results suggest that this protease is potentially useful in many industrial applications.

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Acknowledgments

Thanks to GCUF for the GCUF Research grant program, Project No. 15-ACH-15, and HEC for HEC Project NO. 9512/Punjab/NRPU/R&D/HEC/2017. Consejo Nacional de Ciencia y Tecnología (CONACYT) is thankfully acknowledged for partially supporting this work under Sistema Nacional de Investigadores (SNI) program awarded to Hafiz M.N. Iqbal (CVU: 735340).

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Authors and Affiliations

  1. Department of Biochemistry, Government College University, Faisalabad, 38000, Pakistan

    Shagufta Kamal & Fouzia Hussain

  2. Department of Chemistry, Islamia University, Bahawalpur, 63100, Pakistan

    Ismat Bibi

  3. Department of Botany, Government College University, Faisalabad, 38000, Pakistan

    Muhammad Azeem

  4. Department of Statistics, Government College University, Faisalabad, 38000, Pakistan

    Tanvir Ahmad

  5. Tecnologico de Monterrey, School of Engineering and Sciences, 64849, Monterrey, Mexico

    Hafiz M. N. Iqbal

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  1. Shagufta Kamal
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Kamal, S., Hussain, F., Bibi, I. et al. Mutagenesis and Immobilization of ChitB-Protease for Induced De-staining and Goat Skin Dehairing Potentialities. Catal Lett 152, 12–27 (2022). https://doi.org/10.1007/s10562-021-03605-9

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