title: Adsorption and hydrolysis reactions of poly(hydroxybutyric acid) depolymerases secreted from Ralstonia pickettii T1 and Penicillium funiculosum onto poly[(R)-3-hydroxybutyric acid].
abstract: Reaction processes of poly[(R)-3-hydroxybutyric acid] (P(3HB)) with two types of poly(hydroxybutyric acid) (PHB) depolymerases secreted from Ralstonia pickettii T1 and Penicillium funiculosum were characterized by means of atomic force microscopy (AFM) and quartz crystal microbalance (QCM). The PHB depolymerase from R. pickettii T1 consists of catalytic, linker, and substrate-binding domains, whereas the one from P. funiculosum lacks a substrate-binding domain. We succeeded in observing the adsorption of single molecules of the PHB depolymerase from R. pickettii T1 onto P(3HB) single crystals and the degradation of the single crystals in a phosphate buffer solution at 37 degrees C by real-time AFM. On the contrary, the enzyme molecule from P. funiculosum was hardly observed at the surface of P(3HB) single crystals by real-time AFM, even though the enzymatic degradation of the single crystals was surely progressed. On the basis of the AFM observations in air of the P(3HB) single crystals after the enzymatic treatments, however, not only the PHB depolymerase from R. pickettii T1 but also that from P. funiculosum adsorbed onto the surface of P(3HB) crystals, and both concentrations of the enzymes on the surface were nearly identical. This means both enzymes were adsorbed onto the surface of P(3HB) single crystals. Moreover, QCM measurements clarified quantitatively the differences in detachment behavior between two types of PHB depolymerases, namely the enzyme from R. pickettii T1 was hardly detached but the enzyme from P. funiculosum was released easily from the surface of P(3HB) crystals under an aqueous condition.
authors.affiliation: Department of Innovative and Engineered Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Numata
authors.forename: Koichi
authors.initials: K
authors.lastname: Yamashita
authors.forename: Masahiro
authors.initials: M
authors.lastname: Fujita
authors.forename: Takeharu
authors.initials: T
authors.lastname: Tsuge
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Kasuya
authors.forename: Tadahisa
authors.initials: T
authors.lastname: Iwata
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Doi
authors.forename: Hideki
authors.initials: H
authors.lastname: Abe
mesh_terms: D000217:Acyltransferases / Q000378:metabolism*; D000327:Adsorption; D006868:Hydrolysis; D006885:Hydroxybutyrates / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D010407:Penicillium / Q000201:enzymology*; D011091:Polyesters / Q000378:metabolism*; D043367:Ralstonia / Q000201:enzymology*; D000094122:Polyhydroxybutyrates
chemical_list: D000217:Acyltransferases; D006885:Hydroxybutyrates; D011091:Polyesters; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C060970:poly-beta-hydroxybutyrate polymerase