Microbial sources are regularly used as reliable biocatalysts sources which are often used in the process and production industry. Demands for such organisms with greater capacity of intended enzyme production are on the rise. Lipase is important enzyme used in the biotechnological process of hydrolysis of fats in almost all the relevant industries We have utilized the local oil-contaminated soil resources to search for efficacious bacterial strains that have excellent lipase activity. We were successful in identifying two such bacterial sources, namely, Bacillus subtilis strain RCPS3 and Bacillus fumarioli strain RCPS4, responsible for lipase production from oil effluent contaminated soil of Telangana. This is the first report of these two strains from this part of India that are involved in lipase production. The strains were isolated, optimized, and purified using standard microbiology protocols and were characterized at the molecular level using the biomarker 16s ribosomal RNA genes of the strains. The identified and isolated bacterial strains were confirmed as Bacillus subtilis strain RCPS3, and Bacillus fumarioli strain RCPS4 through molecular and computational characterization.

Identification of proper microbial sources and optimizing the enzyme production conditions are essential for industrial-scale enzyme production. The present study was done to identify and enhance the production of protease enzyme from an important microbial source Rummeliibacillus stabekisii (TWSS-P-2). Ultra-violet radiation physical method and ethyl methanesulfonate and ethidium bromide dependent chemical methods were considered for mutagenesis. Enzyme assay-dependent screening resulted in identifying Rummeliibacillus stabekisii (TWSS-P-2) as the best strain with optimum protease production that was improved through the chemical treatment mentioned. The strains were tested under various physical and chemical factors including carbon source, nitrogen source, inoculum sizes, pH, temperature to optimize the production of the protein. Submerged fermentation (SmF) was used to assess enzyme production. We were successful in deriving the optimum condition for the protease enzyme production for Rummeliibacillus stabekisii (TWSS-P-2) and the mutagenic effect yielded 2-4 fold better enzyme production.

The exploration of novel sources of commercially important enzymes is important due to the enormous industrial requirement. Modern isolation, screening and characterization techniques expedited such efforts. Protease is a highly specific and one of the most important primary enzymes having extensive industrial applications. It is being used in almost all the industrial sectors for one or the other hydrolyzing requirement.The aim of the present study was to isolate, screen, and performing molecular characterization of protease producing novel bacterial strain, sampled from the environmental waste soil around the Warangal region. A total of 25 samples were collected and serial dilution was done with 0.1ml of the sample, further,spreading was done on nutrient agar medium and the sample was incubated at 37⁰C for 24 to 48 hrs.Proteolytic activity in the colony-forming microorganisms was detected through skimmed milk agar medium. The results suggest that samples collected from the tannery waste soil showed high protease activity. A total of 10 colonies of TWSS-P-2 showed high protease activity through larger hydrolysis zone creation (33.5mm).Morphological and biochemical characters were studied based on their isolate and molecular characterization confirmed the strains as Rummelli Bacillus Stabekisii (TWSS-P-2).The maximum activityof the selected strains was observed at pH 7.2 and at temperature 37⁰C.We have conducted several production enhancement and optimization experiments using different carbon and nitrogen sources. Among different carbon and nitrogen sources studied, the highest protease activity of 596(U/ml) was found optimal. Our analysis confirmed that the most effective bacterial strain for the sampled soil was Rummelli Bacillus Stabekisii (WSS-P-2).