TY - JOUR
T1 - Isoeugenol monooxygenase and its putative regulatory gene are located in the eugenol metabolic gene cluster in Pseudomonas nitroreducens Jin1
AU - Ryu, Ji Young
AU - Seo, Jiyoung
AU - Unno, Tatsuya
AU - Ahn, Joong Hoon
AU - Yan, Tao
AU - Sadowsky, Michael J.
AU - Hur, Hor Gil
N1 - 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.
PY - 2010/3
Y1 - 2010/3
N2 - 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.
AB - 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.
KW - Isoeugenol monooxygenase Pseudomonas Vanillin Eugenol Regulator
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U2 - 10.1007/s00203-010-0547-y
DO - 10.1007/s00203-010-0547-y
M3 - Article
C2 - 20091296
AN - SCOPUS:77950360139
SN - 0302-8933
VL - 192
SP - 201
EP - 209
JO - Archives of Microbiology
JF - Archives of Microbiology
IS - 3
ER -