Applied Biochemistry and Biotechnology

, Volume 38, Issue 1, pp 83–92

Salt-tolerant and thermostable alkaline protease fromBacillus subtilis NCIM No. 64


  • Asha A. Kembhavi
    • Division of Biochemical SciencesNational Chemical Laboratory
  • Anuradha Kulkarni
    • Division of Biochemical SciencesNational Chemical Laboratory
  • Aditi Pant
    • Division of Biochemical SciencesNational Chemical Laboratory

DOI: 10.1007/BF02916414

Cite this article as:
Kembhavi, A.A., Kulkarni, A. & Pant, A. Appl Biochem Biotechnol (1993) 38: 83. doi:10.1007/BF02916414


The proteolytic activity produced by aBacillus subtilis isolated from a hot spring was investigated. Maximum protease production was obtained after 38 h of fermentation. Effects of various carbon and nitrogen sources indicate the requirement of starch and bacteriological peptone to be the best inducers for maximum protease production. Requirement for phosphorus was very evident, and the protease was secreted over a wide range of pH 5–11.

The partially purified enzyme was stable at 60°C for 30 min. Calcium ions were effective in stabilizing the enzyme, especially at higher temperature. The enzyme was extremely salt tolerant and retained 100% activity in 5M NaCl over 96 h. The molecular weight of the purified enzymes as determined by SDS-PAGE was 28,000. The enzyme was completely inactivated by PMSF, but little affected by urea, sodium dodecyl sulfate, and sodium tripoly phosphate.

Index Entries

Alkaline proteasesalt and thermostable proteaseBacillus subtilis
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© Humana Press Inc. 1993