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Archives of Microbiology

, Volume 192, Issue 3, pp 201–209 | Cite as

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
Original Paper

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, real-time PCR analyses indicated that the genes involved in the metabolism of eugenol and isoeugenol were differently induced by isoeugenol, eugenol, and vanillin.

Keywords

Isoeugenol monooxygenase Pseudomonas Vanillin Eugenol Regulator 

Notes

Acknowledgment

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

Supplementary material

203_2010_547_MOESM1_ESM.doc (2.6 mb)
Supplementary material 1 (DOC 2648 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Ji-Young Ryu
    • 1
  • Jiyoung Seo
    • 1
  • Tatsuya Unno
    • 1
  • Joong-Hoon Ahn
    • 3
  • Tao Yan
    • 4
  • Michael J. Sadowsky
    • 5
  • Hor-Gil Hur
    • 1
    • 2
  1. 1.Department of Environmental Science and EngineeringGwangju Institute of Science and TechnologyGwangjuKorea
  2. 2.International Environmental Research CenterGwangju Institute of Science and TechnologyGwangjuKorea
  3. 3.Department of Bioscience and BiotechnologyKonkuk UniversitySeoulKorea
  4. 4.Department of Civil and Environmental EngineeringUniversity of HawaiiHonolulu, HawaiiUSA
  5. 5.Department of Soil, Water, and Climate, BioTechnology InstituteUniversity of MinnesotaSt. PaulUSA

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