Purification and characterization of a lactonase from Erwinia cypripedii 314B that hydrolyzes ( S)-5-oxo-2-tetrahydrofurancarboxylic acid.
A bacterium, strain 314B, able to assimilate ( S)-5-oxo-2-tetrahydrofurancarboxylic acid was isolated from soil and identified as Erwinia cypripedii. A lactonase hydrolyzing ( S)-5-oxo-2-tetrahydrofurancarboxylic acid to l-alpha-hydroxyglutaric acid was purified 63-fold with 2% recovery from crude extracts of this bacterium to homogeneity as judged by SDS-PAGE. The molecular masses estimated by SDS-PAGE and gel filtration were 41 kDa and 79 kDa, respectively. The maximum activity was observed at pH 6.5-7.5 and 35-45 degrees C. The enzyme showed lower activity toward dl-2-oxotetrahydrofuran-4,5-dicarboxylic acid, but did not act on ( R)-5-oxo-2-tetrahydrofurancarboxylic acid and other natural and synthetic lactones tested.
Mochizuki
Kazuya
K
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Tsukuba Central 6, 1-1-1 Higashi, 305-8566 Tsukuba, Ibaraki, Japan. mochizuki-kazuya@aist.go.jp
D000595:Amino Acid Sequence; D000645:Ammonium Sulfate; D002413:Cations, Divalent / Q000378:metabolism; D002845:Chromatography / Q000379:methods; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D004885:Erwinia / Q000201:enzymology*; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D005663:Furans / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006868:Hydrolysis; D008670:Metals / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011108:Polymers; D013379:Substrate Specificity; D013696:Temperature
C456614:5-oxo-2-tetrahydrofurancarboxylic acid; D002413:Cations, Divalent; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D005663:Furans; D008670:Metals; D011108:Polymers; C041725:butyl-Toyopearl 650; D017886:Durapatite; D006867:Hydrolases; C460562:Zhd101 protein, Bionectria ochroleuca; D000645:Ammonium Sulfate