Production and Purification of Novel Alkaline Protease Producing Bacillus firmus BAAP-43 isolated from Tannery and Leather Industries Soils

Author: Anusuya Balan and Venkatachalam Palanisamy

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Abstract

Proteolytic enzymes are the enzymes that bring about degradation of the proteins into peptides and amino acids. Proteases enzymes are one of the most important groups of industrial enzymes accounting for more than 65% of the total market for industrial enzymes used in laundry detergent, brewing, the leather, and dairy, pharmaceutical and food industries, and in the production of protein. Protease can hydrolyze the hardly soluble protein on fabric into soluble peptide chain and amino acid in detergent solution. Smoothness. A protease producing microorganism was isolated from tannery and leather industries soil collected from tannery effluent contaminated sites. Based on the 16S rRNA sequencing morphological, biochemical, and molecular characterization the isolate was identified as Bacillus firmus BAAP-43. Maximum protease production was achieved at 45̊C and pH of 9.5 and multiple sources of carbon for its alkaline proteases production; glucose was the best source of carbon, peptone was found to be the best nitrogen source for the maximum enzyme production. Mg2+ enhances the growth and enzyme production and among the different substrates used, casein showed maximum enzyme production. The enzyme was purified by ammonium sulphate precipitation, DEAE- Cellulose and Sephadex G-100 gel chromatography. The Purified protein was with specific activity of 18360.6 U/mg protein was obtained with 43.1 purification fold and 18.7% recovery percentage. Molecular weight of purified enzyme was as 30.2 kDa by SDS-PAGE.

Keywords

Alkaline Protease, Bacillus firmus, Purification, Characterization

Conclusion

In the present study, native alkaline protease producing alkalophilic bacteria was isolated from local tannery and leather industries sediment soil samples as a potent producer of alkaline protease enzyme. The new isolate was identified as Bacillus firmus BAAP-43 based on biochemical tests and molecular identification using 16S rRNA sequence analysis. Culture conditions such as optimum incubation time, temperature, pH, Carbon, Nitrogen sources, Metal ions and substrates for the maximum growth and enzyme production were determined. The molecular weight of the purified alkaline protease was estimated to be 30.2 kDa. Further, a complete large-scale process optimization, purification, and characterization work is needed to achieve inexpensive production of alkaline protease.

References

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How to cite this article

Anusuya Balan and Venkatachalam Palanisamy (2023). Production and Purification of Novel Alkaline Protease Producing Bacillus firmus BAAP-43 isolated from Tannery and Leather Industries Soils. Biological Forum – An International Journal, 15(5): 144-151.