Optimization of culture conditions and properties of immobilized sulfide oxidase from Arthrobacter species.
Arthrobacter species strain FR-3, isolated from sediments of a swamp, produced a novel serine-type sulfide oxidase. The production of sulfide oxidase was maximal at pH 7.5 and 30 degrees C. Among various carbon and nitrogen sources tested, glucose and yeast extract were found to be the most effective substrates for the secretion of sulfide oxidase. The sulfide oxidase was purified to homogeneity and the molecular weight of the purified enzyme was 43 kDa when estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified sulfide oxidase can be effectively immobilized in DEAE (diethylaminoethyl)-cellulose matrix with a yield of 66%. The purified free and immobilized enzyme had optimum activity at pH 7.5 and 6.0, respectively. Immobilization increases the stability of the enzyme with respect to temperature. The half-life of the immobilized enzyme was 30 min at 45 degrees C, longer than that of the free enzyme (10 min). The purified free sulfide oxidase activity was completely inhibited by 1 mM Co2+ and Zn2+ and sulfhydryl group reagents (para-chloromercuribenzoic acid and iodoacetic acid). Catalytic activity was not affected by 1 mM Ca2+, Mg2+, Na+ and metal-chelating agent (EDTA).
Mohapatra
B R
BR
CANMET Materials Technology Laboratory, Natural Resources Canada, 568 Booth Street, Ottawa, Ont., Canada K1A0G1. bmohapat@uvic.ca
Gould
W D
WD
Dinardo
O
O
Papavinasam
S
S
Revie
R W
RW
D001173:Arthrobacter / Q000254:growth & development* / Q000378:metabolism*; D019149:Bioreactors / Q000382:microbiology*; D018929:Cell Culture Techniques / Q000379:methods*; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry*; D011786:Quality Control
D004800:Enzymes, Immobilized; D050862:Oxidoreductases Acting on Sulfur Group Donors; C080499:sulfide oxidase