PURIFICATION AND CHARACTERIZATION OF BACILLUS SUBTILIS JM-3 PROTEASE FROM ANCHOVY SAUCE

Authors: KIM, W.J.; KIM, S.M.

Source: Journal of Food Biochemistry, Volume 29, Number 5, October 2005 , pp. 591-610(20)

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Abstract:

Bacillus subtilis JM-3 was isolated from anchovy sauce naturally fermented in an underground cellar at 15 ± 3C for 3 years. The activity of the B. subtilis protease was highest in the 40–60% ammonium sulfate fraction. The yield of the purified protease was 5.3%, and its purification ratio was 35.6 folds. The molecular weight of the B. subtilis protease was 17.1 kDa, and its K_m and V_maxvalues were 1.75 μg/mL and 318 μM 1/min, respectively. The optimal temperature for protease activity was 60C, but optimal stability temperature was 30C. The optimal pH for protease activity and stability was 5.5. Therefore, the B. subtilis JM-3 protease was classified as an acid protease. The relative activities of the B. subtilis JM-3 protease were 69, 21 and 1.3% at 10, 20 and 30% NaCl concentrations, respectively. The best substrate for the B. subtilis JM-3 protease was benzyloxycarbonyl-glycine-p-nitrophenyl ester followed by bovine serum albumin. p-Toluene-sulfonyl-L-lysine chloromethylketone was the strongest inhibitor followed by soybean trypsin inhibitor, but N-ethylmaleimide did not inhibit this enzyme. The B. subtilis JM-3 protease was therefore presumed to be a trypsin-like serine protease.

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1745-4514.2005.00041.x

Affiliations: 1: Faculty of Marine Bioscience & Technology Kangnung National University Gangneung, Gangwondo 210-702 Korea.

Publication date: 2005-10-01