Purification and characterization of a lactonase from Burkholderia sp. R-711, that hydrolyzes (R)-5-oxo-2-tetrahydrofurancarboxylic acid

• Published in: Archives of microbiology 2001-06, Vol.175 (6), p.430-434
• Main Author: MOCHIZUKI, Kazuya
• Format: Article
• Language: English
• Subjects: Bacteriology; Biological and medical sciences; Burkholderia; Burkholderia - chemistry; Burkholderia - classification; Burkholderia - enzymology; Carboxylic Ester Hydrolases - antagonists & inhibitors; Carboxylic Ester Hydrolases - chemistry; Carboxylic Ester Hydrolases - isolation & purification; Carboxylic Ester Hydrolases - metabolism; Catalysis; Electrophoresis, Polyacrylamide Gel; Enzyme Inhibitors - pharmacology; Fundamental and applied biological sciences. Psychology; Furans - metabolism; Hydrogen-Ion Concentration; Hydrolysis; Kinetics; Lactones - metabolism; Metabolism. Enzymes; Microbiology; Molecular Weight; R-5-oxo-2-tetrahydrofurancarboxlic acid
• ISSN: 0302-8933; 1432-072X
• DOI: 10.1007/s002030100283

A lactonase hydrolyzing (R)-5-oxo-2-tetrahydrofurancarboxylic acid to D-alpha-hydroxyglutaric acid was purified 170-fold with 2% recovery to near homogeneity from crude extracts of Burkholderia sp. R-711, which had been isolated as a bacterium able to assimilate (R)-5-oxo-2-tetrahydrofurancarboxylic acid. The molecular mass was estimated to be 33 kDa by gel filtration. The purified preparation migrated as a single band of molecular mass 38 kDa upon SDS-PAGE. The maximum activity was observed at pH 7.0-8.0 and 35-40 degrees C. The enzyme required no added cofactors or metal ions; the activity was inhibited to 60-100% by SH-blocking reagents, but was not affected by metal-chelating reagents. The enzyme showed lower activity and affinity toward (S)-5-oxo-2-tetrahydrofurancarboxylic acid, but did not act on other natural and synthetic lactones tested.