title: Characterization and functional expression of a rubber degradation gene of a Nocardia degrader from a rubber-processing factory.
abstract: A rubber-degrading bacterial consortium named H2DA was obtained from an enrichment culture with natural rubber latex and rubber-processing factory waste in Vietnam. Gel permeation chromatography analysis revealed that only the strain NVL3 degraded synthetic poly(cis-1,4-isoprene) into low-molecular-weight intermediates among the three strains found in the H2DA. The 16S-rRNA gene sequence of NVL3 showed the highest identity with that of Nocardia farcinica DSM 43665T. NVL3 accumulated aldehyde intermediates from synthetic poly(cis-1,4-isoprene) on a rubber-overlay plate as indicated by Schiff's staining. NVL3 also degraded deproteinized natural rubber into low-molecular-weight aldehyde intermediates. A latex-clearing protein (lcp) gene ortholog was identified within the genome sequence of NVL3, and it showed a moderate amino-acid identity (54-75%) with the lcp genes from previously reported rubber degraders. The heterologous expression of the NVL3 lcp in Escherichia coli BL21(DE3) allowed us to purify the 46.8-kDa His-tagged lcp gene product (His-Lcp). His-Lcp degraded synthetic poly(cis-1,4-isoprene) and accumulated aldehyde intermediates from deproteinized natural rubber suggesting the functional expression of the lcp gene from a Nocardia degrader in E. coli. Quantitative reverse transcription PCR analysis indicated the strong transcriptional induction of the lcp gene in NVL3 in the presence of synthetic poly(cis-1,4-isoprene). These results suggest the involvement of the lcp gene in rubber degradation in NVL3.
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan; School of Biotechnology and Food Technology, Hanoi University of Science and Technology, Hanoi, Viet Nam.
authors.forename: Dao Viet
authors.initials: DV
authors.lastname: Linh
authors.affiliation: School of Biotechnology and Food Technology, Hanoi University of Science and Technology, Hanoi, Viet Nam.
authors.forename: Nguyen Lan
authors.initials: NL
authors.lastname: Huong
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan.
authors.forename: Michiro
authors.initials: M
authors.lastname: Tabata
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Imai
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan.
authors.forename: Sou
authors.initials: S
authors.lastname: Iijima
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kasai
authors.affiliation: School of Biotechnology and Food Technology, Hanoi University of Science and Technology, Hanoi, Viet Nam.
authors.forename: To Kim
authors.initials: TK
authors.lastname: Anh
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan. Electronic address: masao@vos.nagaokaut.ac.jp.
authors.forename: Masao
authors.initials: M
authors.lastname: Fukuda
mesh_terms: D000447:Aldehydes / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D045782:Hemiterpenes / Q000737:chemistry / Q000378:metabolism; D007221:Industry; D007840:Latex / Q000737:chemistry / Q000378:metabolism; D009615:Nocardia / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D012408:Rubber / Q000737:chemistry / Q000378:metabolism*; D014744:Vietnam
chemical_list: D000447:Aldehydes; D045782:Hemiterpenes; D007840:Latex; C455100:1,4-polyisoprene; D012408:Rubber
keywords: Degradation; Functional expression; Latex-clearing protein; Nocardia; Rubber