Isolation and Purification of Dehalogenase From Pseudomonas cepacia UK7WS1 strain

Tatang Hernawan ( Dept. of Pharmacy, Faculty of Mathematics and Natural Sciences, ITB)
S. Soemitro S. Soemitro ( Dept. of Chemistry, Faculty of Mathematics and Natural Sci., UNPAD)
H. Dewayani H. Dewayani ( Dept. of Chemistry, Faculty of Mathematics and Natural Sci., UNPAD)
Tetih Tetih ( IURC on Biotechnology, ITB)
W. T. Ismaya ( IURC on Biotechnology, ITB)

Jurnal Matematika & Sains
Vol 4, No 2 (1999)

Intracellular dehalogenase enzyme had been isolated from Pseudomonas cepacia UK7WS1 strain which was grown in MSM broth containing yeast extract (0.18 g/L) and MCPA (0,02 g/L) and incubated for 22-24 h at 30 oC. Enzyme purification was carried out by ammonium sulphate fractionation, DEAE-cellulose column chromatography and Sephadex G-75 gel filtration column chromatography. Protein purify factor achieved was 12.4 times and protein yield was 10 % w/w of the crude cell extract protein. The enzyme purity of each purification stage was monitored by SDS-polyacrylamide gel electrophoresis and resulted in one protein band for each denatured enzyme (dimeric protein of two identical subunits) and non-denatured enzyme (monomeric protein subunit).