title: Purification, characterization, and cloning of the gene for a biodegradable plastic-degrading enzyme from Paraphoma-related fungal strain B47-9.
abstract: Paraphoma-related fungal strain B47-9 secreted a biodegradable plastic (BP)-degrading enzyme which amounted to 68 % (w/w) of the total secreted proteins in a culture medium containing emulsified poly(butylene succinate-co-adipate) (PBSA) as sole carbon source. The gene for this enzyme was found to be composed of an open reading frame consisting of 681 nucleotides encoding 227 amino acids and two introns. Southern blot analysis showed that this gene exists as a single copy. The deduced amino acid sequence suggested that this enzyme belongs to the cutinase (E.C.3.1.1.74) family; thus, it was named P araphoma-related fungus cutinase-like enzyme (PCLE). It degraded various types of BP films, such as poly(butylene succinate), PBSA, poly(butylene adipate-co-terephthalate), poly(ε-caprolactone), and poly(DL-lactic acid). It has a molecular mass of 19.7 kDa, and an optimum pH and temperature for degradation of emulsified PBSA of 7.2 and 45 °C, respectively. Ca(2+) ion at a concentration of about 1.0 mM markedly enhanced the degradation of emulsified PBSA.
authors.affiliation: National Institute for Agro-Environmental Sciences (NIAES), 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Suzuki
authors.forename: Masako Tsujimoto
authors.initials: MT
authors.lastname: Noguchi
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Shinozaki
authors.forename: Motoo
authors.initials: M
authors.lastname: Koitabashi
authors.forename: Yuka
authors.initials: Y
authors.lastname: Sameshima-Yamashita
authors.forename: Shigenobu
authors.initials: S
authors.lastname: Yoshida
authors.forename: Takeshi
authors.initials: T
authors.lastname: Fujii
authors.forename: Hiroko K
authors.initials: HK
authors.lastname: Kitamoto
mesh_terms: D000272:Adipates / Q000378:metabolism*; D001203:Ascomycota / Q000201:enzymology* / Q000235:genetics; D063566:Biodegradable Plastics / Q000378:metabolism*; D015139:Blotting, Southern; D002118:Calcium / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002413:Cations, Divalent / Q000378:metabolism; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D020536:Enzyme Activators / Q000378:metabolism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007438:Introns; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013386:Succinates / Q000378:metabolism*; D013696:Temperature
chemical_list: D000272:Adipates; D063566:Biodegradable Plastics; D002413:Cations, Divalent; D004271:DNA, Fungal; D020536:Enzyme Activators; D013386:Succinates; C437084:poly(tetramethylene succinate-co-tetramethylene adipate); D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D002118:Calcium