Benzylic and aryl hydroxylations of m-xylene by o-xylene dioxygenase from Rhodococcus sp. strain DK17

Dockyu Kim, Ki Young Choi, Miyoun Yoo, Jung Nam Choi, Choong Hwan Lee, Gerben J. Zylstra, Beom Sik Kang, Eungbin KIm

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Escherichia coli cells expressing Rhodococcus DK17 o-xylene dioxygenase genes were used for bioconversion of m-xylene. Gas chromatography-mass spectrometry analysis of the oxidation products detected 3-methylbenzylalcohol and 2,4-dimethylphenol in the ratio 9:1. Molecular modeling suggests that o-xylene dioxygenase can hold xylene isomers at a kink region between α6 and α7 helices of the active site and α9 helix covers the substrates. m-Xylene is unlikely to locate at the active site with a methyl group facing the kink region because this configuration would not fit within the substrate-binding pocket. The m-xylene molecule can flip horizontally to expose the meta-position methyl group to the catalytic motif. In this configuration, 3-methylbenzylalcohol could be formed, presumably due to the meta effect. Alternatively, the m-xylene molecule can rotate counterclockwise, allowing the catalytic motif to hydroxylate at C-4 yielding 2,4-dimethylphenol. Site-directed mutagenesis combined with structural and functional analyses suggests that the alanine-218 and the aspartic acid-262 in the α7 and the α9 helices play an important role in positioning m-xylene, respectively.

Original languageEnglish
Pages (from-to)1841-1847
Number of pages7
JournalApplied Microbiology and Biotechnology
Volume86
Issue number6
DOIs
Publication statusPublished - 2010 May 1

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Hydroxylation
Catalytic Domain
Dioxygenases
Xylenes
Site-Directed Mutagenesis
Aspartic Acid
Alanine
Gas Chromatography-Mass Spectrometry
3-xylene
Rhodococcus o-xylene dioxygenase
Escherichia coli
Genes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Kim, Dockyu ; Choi, Ki Young ; Yoo, Miyoun ; Choi, Jung Nam ; Lee, Choong Hwan ; Zylstra, Gerben J. ; Kang, Beom Sik ; KIm, Eungbin. / Benzylic and aryl hydroxylations of m-xylene by o-xylene dioxygenase from Rhodococcus sp. strain DK17. In: Applied Microbiology and Biotechnology. 2010 ; Vol. 86, No. 6. pp. 1841-1847.
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abstract = "Escherichia coli cells expressing Rhodococcus DK17 o-xylene dioxygenase genes were used for bioconversion of m-xylene. Gas chromatography-mass spectrometry analysis of the oxidation products detected 3-methylbenzylalcohol and 2,4-dimethylphenol in the ratio 9:1. Molecular modeling suggests that o-xylene dioxygenase can hold xylene isomers at a kink region between α6 and α7 helices of the active site and α9 helix covers the substrates. m-Xylene is unlikely to locate at the active site with a methyl group facing the kink region because this configuration would not fit within the substrate-binding pocket. The m-xylene molecule can flip horizontally to expose the meta-position methyl group to the catalytic motif. In this configuration, 3-methylbenzylalcohol could be formed, presumably due to the meta effect. Alternatively, the m-xylene molecule can rotate counterclockwise, allowing the catalytic motif to hydroxylate at C-4 yielding 2,4-dimethylphenol. Site-directed mutagenesis combined with structural and functional analyses suggests that the alanine-218 and the aspartic acid-262 in the α7 and the α9 helices play an important role in positioning m-xylene, respectively.",
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Benzylic and aryl hydroxylations of m-xylene by o-xylene dioxygenase from Rhodococcus sp. strain DK17. / Kim, Dockyu; Choi, Ki Young; Yoo, Miyoun; Choi, Jung Nam; Lee, Choong Hwan; Zylstra, Gerben J.; Kang, Beom Sik; KIm, Eungbin.

In: Applied Microbiology and Biotechnology, Vol. 86, No. 6, 01.05.2010, p. 1841-1847.

Research output: Contribution to journalArticle

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AU - Choi, Jung Nam

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AU - Zylstra, Gerben J.

AU - Kang, Beom Sik

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