Isolation, Screening, Characterization, and Identification of Alkaline Protease Producing Bacteria from the Dairy Industrial Soil

Author: Merlin Jeena Kumari M. and Sasi Premila J.M.

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Abstract

The purpose of the present study was to isolate, screen and identify protease producing bacteria from two dairy industry soil located in Sankarankoil, Tenkasi district, Tamilnadu, India-627761. The samples are collected and stored based on the microbial standard protocol. Serial dilution and skim agar media are followed for the primary screening to isolate protease producing bacteria. Among twenty isolated microbial cultures two significant organism JKSP1 and MJSP2 are selected based on the highest protease activity zone. The morphological and biochemical tests reveal the isolated species was found to be Bacillus species. Molecular identification was also carried out for the isolated species using 16s RNA sequencing and data are submitted to the gene bank with the accession number (OK605054.1) and (ON994262.1). The BLAST similarity shows that isolate JKSP1 is 100% homologous to Bacillus sp strain (OQ071600.1) and MJSP2 100% homologous to Bacillus subtilis (ON994262.1). Additionally, optimizing protease production on a large scale remains a difficult task. Hence this study mainly focused on enzyme production optimization and characterization with respect to pH, temperature, incubation time and different substrates like metal iron, carbon, nitrogen sources and agro-bases. The optimization results indicate that; the isolated organism MJSP2 shows promising production of enzymes at minimum resources and physical parameters. The overall outcome concluded MJSP2 strains show efficient proteolysis activity and can be considered an effective protease producer for commercial value.

Keywords

Bacteria, Bacillus sp, Bacillus subtilis, Protease, Soil, Optimization

Conclusion

The present study was focused on isolating, screening, and identification of protease producing bacteria from dairy industry soil samples. Among 20 isolated samples, two bacterial isolates showed potent protease activity on skim milk agar plate assay. The bacterial isolates were then further screened for alkaline protease production using the submerged fermentation method and the isolated MJSP2 was observed to have a maximum protease enzyme. Based on morphological and biochemical characteristics the strain was identified as Bacillus species which was later identified as Bacillus sp strain and Bacillus subtilis species using 16S rRNA sequencing techniques.

References

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How to cite this article

Merlin Jeena Kumari M. and Sasi Premila J.M. (2023). Isolation, Screening, Characterization, and Identification of Alkaline Protease Producing Bacteria from the Dairy Industrial Soil. Biological Forum – An International Journal, 15(2): 35-41.