Abstract:
Burkholderia stabilis, a protease producing organism was isolated from green house soils in
Chungchugnam-do province, Gongju-Gun area in South Korea. Optimum protease activity of 116.4 U/ml
was observed in the growth medium containing 0.7% KH2PO4, 0.2% K2HPO4, 0.01% MgSO4.7H2O, 0.05%
citric acid dehydrate, 0.1% yeast extract and 0.2% casein. The protease production was found to be
optimized in 1: 5 cultivation volume with 1% inoculum, shaken at 150 rpm. The enzyme was active in pH
range 5 to11 and temperature of 30 to 80°C. The optimum pH and the temperature for protease activity
were recorded to be pH 8 and 50°C, respectively. The enzyme was stable up to 40°C and pH 9. The
protease activity was inhibited by Zn2+, Ni2+ and Sn2+ and increased by Ca
2+, Mg2+ and Mn2+. The
maximum enzyme activity was displayed with casein as the substrate followed by egg albumin, gelatin
and bovine serum albumin (BSA). Vmax and Km values were 89.28 U/ml and 0.82 mg/ml, respectively
when casein was a substrate. The protease was purified to homogeneity by a combination of
ammonium sulphate precipitation and gel filtration chromatography. The molecular weight of the
enzyme was recorded as 45 kDa by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis.
