Exploration of Compost Soil for the Production of Thermo-Stable <i>Bacillus</i> Protease to Synthesize Bioactive Compounds through Soy Protein Hydrolysis
Ieshita Pan,
Krishnamoorthy Nanjundan,
Aravindan Achuthan,
Praveen Kumar Issac,
Rajinikanth Rajagopal,
Soon Woong Chang,
Sartaj Ahmad Bhat,
Balasubramani Ravindran
Affiliations
Ieshita Pan
Department of Medical Biotechnology and Integrative Physiology, Institute of Biotechnology, Saveetha School of Engineering, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai 602105, Tamil Nadu, India
Krishnamoorthy Nanjundan
Department of Physics, Sri Eshwar College of Engineering, Kinathukadavu, Coimbatore 641202, Tamil Nadu, India
Aravindan Achuthan
Department of Civil Engineering, Koneru Lakshmaiah Education Foundation, Green Field, Vaddeswaram, Guntur 522302, Andhra Pradesh, India
Praveen Kumar Issac
Department of Medical Biotechnology and Integrative Physiology, Institute of Biotechnology, Saveetha School of Engineering, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai 602105, Tamil Nadu, India
Rajinikanth Rajagopal
Sherbrooke Research and Development Center, Agriculture and Agri-Food Canada, 2000 College Street, Sherbrooke, QC J1M 1Z3, Canada
Soon Woong Chang
Department of Environmental Energy & Engineering, Kyonggi University, Suwon-si 16227, Republic of Korea
Sartaj Ahmad Bhat
River Basin Research Center, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan
Balasubramani Ravindran
Department of Environmental Energy & Engineering, Kyonggi University, Suwon-si 16227, Republic of Korea
Application of bioactive peptides (BAPs) is promising due to their potential antimicrobial, antioxidant, agonistic, and ACE inhibition properties. To achieve a stable and active peptide at relatively high pH and temperatures by microbial fermentation, a wide variety of microorganisms need to be explored from diverse habitats, and compost is the excellent source. In an attempt to isolate potent protease-producing bacteria, gelatin-supplemented DM agar medium was used. Out of 140 pure cultures, initial protease production selects isolate D3L/1 (26 U/mL), and 16S rDNA sequencing confirmed it as Bacillus subtilis. Protease production was increased to 55.55 U/mL, with pH 7.5, 1% glucose, 1% casein, 1% ammonium sulfate, for 96 h of fermentation, at 37 °C under 140 rpm of shaking. Ion-exchange, and size-exclusion chromatography, 30 KDa protease was purified up to 4.1-fold (specific activity 3448.62 U/mL; 67.66% yield). The enzyme was active under broad temperatures (60 °C optimum), organic solvents, and pH variations. A total of 5% H2O2 can only reduce 40% of enzyme activity. However, 1 mM, Fe2+, and Cu2+ increased enzyme activity by five times. Soy hydrolysis (SPI) byD3L/1 protease produces bioactive compound (Serratia marcescens but active against Escherechia coli (47%), Staphylococcus aureus (28%), and Pseudomonas aeruginosa (12%).
