Purification, characterization and application of collagenolytic protease from Bacillus subtilis strain MPK

J Biosci Bioeng. 2024 Jul;138(1):21-28. doi: 10.1016/j.jbiosc.2024.03.003. Epub 2024 Apr 17.

Abstract

A new extracellular protease from Bacillus subtilis strain MPK with collagenolytic activity was isolated and purified. Fish skin which otherwise would be treated as waste is used as substrate for the production of protease. Using various techniques such as ammonium sulphate precipitation and ion exchange chromatography, protease was purified and characterized subsequently. Protease of approximately 61 kDa molecular weight was purified by 135.7-fold with 18.42% enzyme recovery. The protease showed effective properties like pH and temperature stability over a broad range with optimum pH 7.5 and temperature 60 °C. Km and Vmax were found to be 1.92 mg ml-1 and 1.02 × 10-4 mol L-1 min-1, respectively. The protease exhibited stability in various ions, surfactants, inhibitors and organic solvents. Subsequently, the protease was successfully utilized for collagen hydrolysis to generate collagen peptides; thus, the produced protease would be a potential candidate for multifaceted applications in food and pharmaceutical industries due to its significant characteristics and collagenolytic properties.

Keywords: Bacillus subtilis strain MPK; Collagen peptides; Collagenolytic protease; Prot MPK.

MeSH terms

  • Animals
  • Bacillus subtilis* / enzymology
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism
  • Chromatography, Ion Exchange
  • Collagen* / chemistry
  • Collagen* / metabolism
  • Enzyme Stability
  • Fishes / microbiology
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Molecular Weight*
  • Peptide Hydrolases / chemistry
  • Peptide Hydrolases / isolation & purification
  • Peptide Hydrolases / metabolism
  • Skin
  • Temperature*

Substances

  • Collagen
  • Peptide Hydrolases
  • Bacterial Proteins