期刊
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
卷 123, 期 4, 页码 412-418出版社
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2016.11.012
关键词
Rubber; Degradation; Nocardia; Latex-clearing protein; Functional expression
资金
- Science and Technology Research Partnership for Sustainable Development (SATREPS)
- Japan Science and Technology Agency (JST)
- Japan International Cooperation Agency (JICA)
- Ministry of Science and Technology of Vietnam
- Ministry of Education and Training of Vietnam
- Grants-in-Aid for Scientific Research [15H05639] Funding Source: KAKEN
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 43665(T). 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 (Icp) gene ortholog was identified within the genome sequence of NVL3, and it showed a moderate amino-acid identity (54-75%) with the Icp genes from previously reported rubber degraders. The heterologous expression of the NVL3 Icp in Escherichia coli BL21(DE3) allowed us to purify the 46.8-kDa His-tagged Icp 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 Icp 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.
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