A review on Enzyme Production, Optimization and Assay methods from Microorganisms- with a focus on peptidases

Abstract

Peptidases, a subclass of enzymes, play an essential role in protein degradation by cleaving peptide bonds based on this they are into six types based on their catalytic mechanisms. They are crucial for various cellular processes, including protein turnover, cell signalling, DNA repair, immune defence, and apoptosis, and are widely studied for their applications in biopolymer degradation, medical treatments, and digestive processes. The Plackett-Burman Design (PBD) is an experimental approach used to efficiently screen multiple variables with minimal experimental runs. PBD has been applied in numerous enzyme production studies, including the optimization of culture media and physical parameters, such as pH, temperature, and inoculum concentration, to improve enzyme yield. Microorganisms such as Aspergillus niger, Lactobacillus amylophilus, and Bacillus species are commonly employed for the production of peptidases. These organisms can produce both intracellular and extracellular enzymes, with the extraction method varying accordingly. Each enzyme type is extracted and quantified using various assays to determine enzyme activity. The assay methods for determining the activity of proteolytic enzymes, collagenase, and glutamyl endopeptidase includes spectrophotometry, colorimetric assays, AMC fluorometry, and thin-layer chromatography were employed. Additionally, enzyme localization was studied by analyzing cell fractions after lysing cells.