Degradation of poly(3-hydroxybutyrate) by poly(3-hydroxybutyrate) depolymerase from Alcaligenes faecalis T1.
The extracellular poly(3-hydroxybutyrate) depolymerase purified from Alcaligenes faecalis T1 has two disulfide bonds, one of which appears to be necessary for the full enzyme activity. This depolymerase hydrolyzed not only hydrophobic poly(3-hydroxybutyrate) but also water-soluble trimer and larger oligomers of D-(-)-3-hydroxybutyrate, regardless of their solubilities in water. Kinetic analyses with oligomers of various sizes indicated that the substrate cleaving site of the enzyme consisted of four subsites with individual affinities for monomer units of the substrate. Analyses of the hydrolytic products of oligomers, which had labeled D-(-)-3-hydroxybutyrate at the hydroxy terminus, showed that the enzyme cleaved only the second ester linkage from the hydroxy terminus of the trimer and tetramer, and acted as an endo-type hydrolase toward the pentamer and higher oligomers. The enzyme appeared to have a hydrophobic site which interacted with poly(3-hydroxybutyrate) and determined the affinity of the enzyme toward the hydrophobic substrate.
D000421:Alcaligenes / Q000201:enzymology*; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006885:Hydroxybutyrates / Q000378:metabolism; D007700:Kinetics; D017646:Organosilicon Compounds; D011091:Polyesters; D011092:Polyethylene Glycols / Q000494:pharmacology; D011108:Polymers / Q000378:metabolism; D012825:Silicon / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D000094122:Polyhydroxybutyrates
D002265:Carboxylic Ester Hydrolases; D006885:Hydroxybutyrates; D017646:Organosilicon Compounds; D011091:Polyesters; D011092:Polyethylene Glycols; D011108:Polymers; D012825:Silicon; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; C009660:t-butyldimethylsilyl compounds; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C022417:poly-beta-hydroxybutyrate depolymerase