Oxidant and SDS-stable alkaline protease from a halo-tolerant Bacillus clausii I-52: enhanced production and simple purification.
OBJECTIVE
An investigation was carried out on the enhancement of protease production and simple purification of an oxidant and SDS-stable alkaline protease produced by Bacillus clausii I-52 of industrial significance.


RESULTS
The supplementation with 0.4% (w/v) NaCl and 0.05% (w/v) FeSO4.7H2O in a culture medium caused an increase in the protease production. The enzyme was purified to homogeneity with overall recovery of 79% and 10-fold purification from culture supernatant using Diaion HPA75, phenyl-Sepharose and DEAE-Sepharose column chromatographies. The protease was a halo-tolerant enzyme with apparent molecular mass of 28 kDa, and the Km and kcat values for N-Succinyl-Ala-Ala-Pro-Phe-pNA at 45 degrees C and pH 11.0 were determined to be 83.9 micromol l(-1) and 238.6 s(-1) respectively.


CONCLUSIONS
Bacillus clausii I-52 was identified as a halo-tolerant bacterium, and the extracellular alkaline protease produced by B. clausii I-52 also showed extreme halo-tolerance. The enzyme stability towards SDS and H2O2 could be increased by adding NaCl or propylene glycol to the enzyme solution.


CONCLUSIONS
The alkaline protease secreted by B. clausii I-52 is significant from an industrial perspective because of its stability against surfactants and oxidants as well as its tolerance towards high salinity. These enzymatic properties suggest its suitable application for industrial purposes.
Joo
H-S
HS
Department of Biochemistry, College of Medicine, Inha University, Chung-Ku, Inchon, Korea.
Chang
C-S
CS
D001407:Bacillus / Q000378:metabolism*; D019149:Bioreactors; D004795:Enzyme Stability; D006861:Hydrogen Peroxide; D007218:Industrial Microbiology; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D012965:Sodium Chloride; D013379:Substrate Specificity; D013501:Surface-Active Agents / Q000494:pharmacology
D016877:Oxidants; D013501:Surface-Active Agents; D012965:Sodium Chloride; D006861:Hydrogen Peroxide; D012697:Serine Endopeptidases; C032923:microbial serine proteinases