Purification and characterization of heat-tolerant protease produced by Bacillus polyfermenticus SCD.
A protease produced by Bacillus polyfermenticus SCD was purified and characterized as a new detergent material. The protease was purified from supernatant produced by B. polyfermenticus SCD, by ammonium sulfate precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, and finally gel filtration chromatography on Sephadex G-50. The molecular mass of this enzyme was 44 kDa based on SDS-PAGE. The optimum temperature and pH were 50°C and pH 8.0. The ranges of its stability to the pH and temperature were 7.0 to 9.0 and under 40°C, respectively. The enzyme was highly stable in the presence of the surfactants like Triton X-100 (0.1%), showing a 2-fold increase in its proteolytic activity. However, the enzyme was slightly inhibited by the chelating agent EDTA (1 mM). The enzyme has a maximum activity at 50°C and the activity can be increased by surfactants such as Triton X-100 and Tween 80.
Division of Animal Life Science, Konkuk University, Seoul 143-701, Republic of Korea.
Gooi Hun
GH
Choi
Mi Na
MN
Jo
Jin-Man
JM
Kim
Cheon-Jei
CJ
Kim
Kee-Tae
KT
Kim
Hyun-Dong
HD
Paik
D001407:Bacillus / Q000201:enzymology*; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D013696:Temperature
D010447:Peptide Hydrolases