Isoeugenol monooxygenase and its putative regulatory gene are located in the eugenol metabolic gene cluster in Pseudomonas nitroreducens Jin1

Ji Young Ryu, Jiyoung Seo, Tatsuya Unno, Joong Hoon Ahn, Tao Yan, Michael J. Sadowsky, Hor Gil Hur

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

The plant-derived phenylpropanoids eugenol and isoeugenol have been proposed as useful precursors for the production of natural vanillin. Genes involved in the metabolism of eugenol and isoeugenol were clustered in region of about a 30 kb of Pseudomonas nitroreducens Jin1. Two of the 23 ORFs in this region, ORFs 26 (iemR) and 27 (iem), were predicted to be involved in the conversion of isoeugenol to vanillin. The deduced amino acid sequence of isoeugenol monooxygenase (Iem) of strain Jin1 had 81.4% identity to isoeugenol monooxygenase from Pseudomonas putida IE27, which also transforms isoeugenol to vanillin. Iem was expressed in E. coli BL21(DE3) and was found to lead to isoeugenol to vanillin transformation. Deletion and cloning analyses indicated that the gene iemR, located upstream of iem, is required for expression of iem in the presence of isoeugenol, suggesting it to be the iem regulatory gene. Reverse transcription, realtime PCR analyses indicated that the genes involved in the metabolism of eugenol and isoeugenol were differently induced by isoeugenol, eugenol, and vanillin.

Original languageEnglish (US)
Pages (from-to)201-209
Number of pages9
JournalArchives of Microbiology
Volume192
Issue number3
DOIs
StatePublished - Mar 2010

Bibliographical note

Funding Information:
Acknowledgment This study was carried out with the support of “On-Site Cooperative Agriculture Research Project (project no. 20070101080021)”, RDA, Suwon, Republic of Korea.

Keywords

  • Isoeugenol monooxygenase Pseudomonas Vanillin Eugenol Regulator

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