caa a22 4500 605501092 CHVBK 20210128100559.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6756-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6756-1 Molecular weight-dependent degradation of d -lactate-containing polyesters by polyhydroxyalkanoate depolymerases from Variovorax sp. C34 and Alcaligenes faecalis T1 [Elektronische Daten] [Jian Sun, Ken'ichiro Matsumoto, Yuta Tabata, Ryosuke Kadoya, Toshihiko Ooi, Hideki Abe, Seiichi Taguchi] Polyhydroxyalkanoate depolymerase derived from Variovorax sp. C34 (PhaZVs) was identified as the first enzyme that is capable of degrading isotactic P[67mol% (R)-lactate(LA)-co-(R)-3-hydroxybutyrate(3HB)] [P(d-LA-co-d-3HB)]. This study aimed at analyzing the monomer sequence specificity of PhaZVs for hydrolyzing P(LA-co-3HB) in comparison with a P(3HB) depolymerase from Alcaligenes faecalis T1 (PhaZAf) that did not degrade the same copolymer. Degradation of P(LA-co-3HB) by action of PhaZVs generated dimers, 3HB-3HB, 3HB-LA, LA-3HB, and LA-LA, and the monomers, suggesting that PhaZVs cleaved the linkages between LA and 3HB units and between LA units. To provide a direct evidence for the hydrolysis of these sequences, the synthetic methyl trimers, 3HB-3HB-3HB, LA-LA-3HB, LA-3HB-LA, and 3HB-LA-LA, were treated with the PhaZs. Unexpectedly, not only PhaZVs but also PhaZAf hydrolyzed all of these substrates, namely PhaZAf also cleaved LA-LA linkage. Considering the fact that both PhaZs did not degrade P[(R)-LA] (PDLA) homopolymer, the cleavage capability of LA-LA linkage by PhaZs was supposed to depend on the length of the LA-clustering region in the polymer chain. To test this hypothesis, PDLA oligomers (6 to 40mer) were subjected to the PhaZ assay, revealing that there was an inverse relationship between molecular weight of the substrates and their hydrolysis efficiency. Moreover, PhaZVs exhibited the degrading activity toward significantly longer PDLA oligomers compared to PhaZAf. Therefore, the cleaving capability of PhaZs used here toward the d-LA-based polymers containing the LA-clustering region was strongly associated with the substrate length, rather than the monomer sequence specificity of the enzyme. Springer-Verlag Berlin Heidelberg, 2015 Biobased polyester nationallicence Polylactic acid nationallicence Biodegradation nationallicence Biodegradable material nationallicence Size-dependent hydrolysis nationallicence Monomer sequence specificity nationallicence Sun Jian Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut Matsumoto Ken'ichiro Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut Tabata Yuta Department of Innovative and Engineering Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8502, Yokohama, Kanagawa, Japan aut Kadoya Ryosuke Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut Ooi Toshihiko Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut Abe Hideki Department of Innovative and Engineering Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8502, Yokohama, Kanagawa, Japan aut Taguchi Seiichi Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9555-9563 0175-7598 99:22<9555 2015 99 253 https://doi.org/10.1007/s00253-015-6756-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6756-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sun Jian Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut NATIONALLICENCE 700 1- Matsumoto Ken'ichiro Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut NATIONALLICENCE 700 1- Tabata Yuta Department of Innovative and Engineering Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8502, Yokohama, Kanagawa, Japan aut NATIONALLICENCE 700 1- Kadoya Ryosuke Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut NATIONALLICENCE 700 1- Ooi Toshihiko Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut NATIONALLICENCE 700 1- Abe Hideki Department of Innovative and Engineering Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8502, Yokohama, Kanagawa, Japan aut NATIONALLICENCE 700 1- Taguchi Seiichi Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, 060-8628, Sapporo, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9555-9563 0175-7598 99:22<9555 2015 99 253