title: Studies on the biodegradability of polythioester copolymers and homopolymers by polyhydroxyalkanoate (PHA)-degrading bacteria and PHA depolymerases.
abstract: The biodegradability of microbial polythioesters (PTEs), a novel class of biopolymers which were discovered recently and can be produced by polyhydroxyalkanoate (PHA)-accumulating bacteria, was studied. Using poly(3-hydroxybutyrate- co-3-mercaptopropionate) [poly(3HB- co-3MP)] as sole carbon source for screening, 22 new bacterial strains were isolated and characterized. Interestingly, none of the PHA-degrading bacteria was able to utilize the homopolymer poly(3MP) as a carbon source for growth or to form clear zones on poly(3MP)-containing agar plates. The extracellular PHA depolymerases from two strains ( Schlegelella thermodepolymerans, Pseudomonas indica K2) were purified to electrophoretic homogeneity and biochemically characterized. The PHA depolymerase of S. thermodepolymerans exhibited a temperate optimum of about 75 degrees C to 80 degrees C and was stable at 70 degrees C for more than 24 h. Regarding the substrate specificities of the PHA depolymerase of S. thermodepolymerans, enzyme activities decreased significantly with increasing 3MP content of the copolymer substrates. Interestingly, no activity could be detected with homoPTEs consisting only of 3MP or of 3-mercaptobutyrate. Similar results were obtained with the PHA depolymerases PhaZ2, PhaZ5 and PhaZ7 of Paucimonas lemoignei which were also investigated. The PHA depolymerase of Ps. indica K2 did not cleave any of the investigated polymers containing 3MP. Gas chromatography, infrared and (1)H-NMR spectrometry and matrix-assisted laser desorption/ionization time-of-flight analysis revealed that 3MPs containing oligomers were enriched in the water-insoluble fraction remaining after partial digestion of poly(3HB- co-3MP) by purified poly(3HB) depolymerase of S. thermodepolymerans. In contrast, 3HB was enriched in the water-soluble fraction, which also contained 3HB- co-3MP dimer obtained by partial digestion of this copolymer by the enzyme. This study clearly indicates that PHA depolymerases are specific for oxoester linkages of PHAs and that the thioester bonds of PTEs cannot be cleaved by this type of enzyme.
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstrasse 3, 48149 Münster, Germany.
authors.forename: Khaled
authors.initials: K
authors.lastname: Elbanna
authors.forename: Tina
authors.initials: T
authors.lastname: Lütke-Eversloh
authors.forename: Dieter
authors.initials: D
authors.lastname: Jendrossek
authors.forename: Heinrich
authors.initials: H
authors.lastname: Luftmann
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinbüchel
mesh_terms: D001419:Bacteria / Q000145:classification / Q000254:growth & development / Q000302:isolation & purification* / Q000378:metabolism*; D020562:Betaproteobacteria / Q000201:enzymology; D001673:Biodegradation, Environmental; D001704:Biopolymers / Q000378:metabolism*; D042521:Burkholderiaceae / Q000201:enzymology; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D006885:Hydroxybutyrates / Q000032:analysis; D011091:Polyesters / Q000032:analysis / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology; D012722:Sewage / Q000382:microbiology; D013379:Substrate Specificity; D013696:Temperature; D014871:Water Microbiology; D000094122:Polyhydroxybutyrates
chemical_list: D001704:Biopolymers; D002265:Carboxylic Ester Hydrolases; D006885:Hydroxybutyrates; D011091:Polyesters; D012722:Sewage; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C077226:poly(3-hydroxyalkanoic acid) depolymerase