• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1169-1176
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• 2007

Genes encoding proteins responsible for limonene catabolism were cloned from a limonene-degrading microorganism, Enterobacter cowanii 6L, which was isolated from citron (Citrus junos) peel. The 8.6, 4.7, and 7.7 kb fragments (CD3, CD4, and CD6) of E. cowanii 6L chromosomal DNA that confer to E. coli the ability to grow on limonene have been cloned and their corresponding DNA sequences were determined. Nine open reading frames (ORFs) were identified, and the four ORFs (921 bp of CD3-2; 1,515 bp of CD4-1; 1,776 bp of CD6-1; and 1,356 bp of CD6-2) that encode limonene hydroxylase were confirmed by independently expressing these genes in E. coli. FAD and NADH were found to stimulate the hydroxylation reaction if added to cell extracts from E. coli recombinants, and multiple compounds (linalool, dihydrolinalool, perillyl alcohol, (${\alpha}-terpineol$, and ${\gamma}-terpineol$) were the principal products observed. Our results suggest that the isolate E. cowanii 6L has a broad metabolic capability including utilization of limonene. This broad metabolic ability was confirmed by identifying four novel limonene hydroxylase functional ORFs in E. cowanii 6L.

• Journal of Microbiology and Biotechnology
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• 제22권7호
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• pp.917-922
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• 2012

Homology modeling of Streptomyces peucetius CYP147F1 was constructed using three cytochrome P450 structures, CYP107L1, CYPVdh, and CYPeryF, as templates. The lowest energy SPCYP147F1 model was then assessed for stereochemical quality and side-chain environment by Accelrys Discovery Studio 3.1 software. Further activesite optimization of the SPCYP147F1 was performed by molecular dynamics to generate the final SPCYP147F1 model. The substrate limonene was then docked into the model. The model-limonene complex was used to validate the active-site architecture, and functionally important residues within the substrate recognition site were identified by subsequent characterization of the secondary structure. The docking of limonene suggested that SPCYP147F1 would have broad specificity with the ligand based on the two different orientations of limonene within the active site facing to the heme. Limonene with C7 facing the heme with distance of $3.4{\AA}$ from the Fe was predominant.

• Applied Biological Chemistry
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• 제36권5호
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• pp.358-363
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• 1993

박하(Mentha piperita) 세포현탁배양에(-)-menthol 생합성 중간체를 투여하여 배양된 세포의 대사경로를 연구하였다. (-)-Limonene을 투여 하였을 때 이는 다른 대사물로 변환되지 않는 것으로 관찰 되었다. (+)-Pulegone은 (+)-isomenthone 및 (-)-menthone으로 변환되었으며, (-)-menthone은 (-)-menthol로 변환되었다. 이 실험은 현탁배양세포가 대부분의 생합성 활성을 유지하고 있으며 (-)-limonene hydroxylase의 활성이 제한적임을 보여 주었다. (-)-Isopiperitenone을 투여하였을 때는 (+)-pulegone, piperitenone, (-)-7-hydroxyisopiperitenone, (R)- 및 (S)-6-hydroxyisopiperitenone이 생성되었다.