Purification and characterization of a novel esterase promising for the production of useful compounds from Microbacterium sp. 7-1W.
A novel esterase catalyzing regioselective hydrolysis was purified from the membrane fraction of Microbacterium sp. 7-1W, and characterized. The enzyme was solubilized with Brij 58 and purified 13.8-fold to apparent homogeneity with 2.58% overall recovery. The relative molecular mass of the native enzyme as estimated by gel filtration was more than 600,000 Da, and the subunit molecular mass was 62,000 Da. The enzyme catalyzed cleavage of the terminal ester bonds of cetraxate esters and pantothenate esters. The K(m) and V(max) values for methyl cetraxate were 0.380 mM and 7.76 micromole min(-1) mg(-1) protein, respectively. The enzyme was inhibited by serine hydrolase inhibitors.
Honda
Kohsuke
K
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
Kataoka
Michihiko
M
Ono
Hiroko
H
Sakamoto
Keiji
K
Kita
Shinji
S
Shimizu
Sakayu
S
D000192:Actinomycetales / Q000201:enzymology*; D004795:Enzyme Stability; D004950:Esterases / Q000302:isolation & purification* / Q000378:metabolism; D004952:Esters / Q000378:metabolism; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D010205:Pantothenic Acid / Q000031:analogs & derivatives; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity; D014148:Tranexamic Acid / Q000031:analogs & derivatives* / Q000737:chemistry
D004952:Esters; D008565:Membrane Proteins; D010205:Pantothenic Acid; C000665:cetraxate; D014148:Tranexamic Acid; D004950:Esterases