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Characterization of high-yield Bacillus subtilis cysteine protease for diverse industrial applications

  • Biotechnology and Industrial Microbiology - Research Paper
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Abstract 

Bacterial proteases have extensive applications in various fields of industrial microbiology. In this study, protease-producing organisms were screened on skimmed milk agar media using serial dilution. Through microbial biomass production, biochemical tests, protease-specific activity, and 16 s RNA gene sequencing, the isolates were identified as Bacillus subtilis and submitted to NCBI. The strain accession numbers were designated as A1 (MT903972), A2 (MT903996), A4 (MT904091), and A5 (MT904796). The strain A4 Bacillus subtilis showed highest protease-specific activity as 76,153.84 U/mg. A4 Bacillus subtilis was unaffected by Ca2+, Cu2+, Fe2+, Hg2+, Mg2+, Na, Fe2+, and Zn2+ but was inhibited by 80% by Mn2+ (5 mM). The protease activity was inhibited by up to 30% by iodoacetamide (5 mM). These findings confirm the enzyme to be a cysteine protease which was further confirmed by MALDI-TOF. The identified protease showed 71% sequence similarity with Bacillus subtilis cysteine protease. The crude cysteine protease significantly aided in fabric stain removal when added to a generic detergent. It also aided in the recovery of silver from used X-ray films and de-hairing of goat skin hides and showed decent application in meat tenderization. Thus, the isolated cysteine protease has high potential for industrial applications.

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Data availability

The data that support the findings of this study are available on request from the corresponding author, SB.

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Funding

This work has been partially funded by OU-DST-PURSE intramural grant and RUSA 2.0.

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Authors and Affiliations

  1. Department of Microbiology, University College of Science, Osmania University, Tarnaka, Hyderabad, 500 007, Telangana, India

    Usha Rani Keshapaga, Kalyani Jathoth & Sandeepta Burgula

  2. Department of Biochemistry , University College of Science, Osmania University , Tarnaka, Hyderabad, 500 007, Telangana, India

    Surya Satyanarayana Singh & Raghu Gogada

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  1. Usha Rani Keshapaga
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Correspondence to Sandeepta Burgula.

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Keshapaga, U.R., Jathoth, K., Singh, S.S. et al. Characterization of high-yield Bacillus subtilis cysteine protease for diverse industrial applications. Braz J Microbiol 54, 739–752 (2023). https://doi.org/10.1007/s42770-023-00992-6

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