Abstract:
:Pseudomonas sp. VLB120 uses styrene as a sole source of carbon and energy. The first step in this metabolic pathway is catalyzed by an oxygenase (StyA) and a NADH-flavin oxidoreductase (StyB). Both components have been isolated from wild-type Pseudomonas strain VLB120 as well as from recombinant Escherichia coli. StyA from both sources is a dimer, with a subunit size of 47 kDa, and catalyzes the enantioselective epoxidation of CC double bonds. Styrene is exclusively converted to S-styrene oxide with a specific activity of 2.1 U mg(-1) (k(cat) = 1.6 s(-1)) and K(m) values for styrene of 0.45 +/- 0.05 mM (wild type) and 0.38 +/- 0.09 mM (recombinant). The epoxidation reaction depends on the presence of a NADH-flavin adenine dinucleotide (NADH-FAD) oxidoreductase for the supply of reduced FAD. StyB is a dimer with a molecular mass of 18 kDa and a NADH oxidation activity of 200 U mg(-1) (k(cat) [NADH] = 60 s(-1)). Steady-state kinetics determined for StyB indicate a mechanism of sequential binding of NADH and flavin to StyB. This enzyme reduces FAD as well as flavin mononucleotide and riboflavin. The NADH oxidation activity does not depend on the presence of StyA. During the epoxidation reaction, no formation of a complex of StyA and StyB has been observed, suggesting that electron transport between reductase and oxygenase occurs via a diffusing flavin.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Otto K,Hofstetter K,Röthlisberger M,Witholt B,Schmid Adoi
10.1128/JB.186.16.5292-5302.2004keywords:
subject
Has Abstractpub_date
2004-08-01 00:00:00pages
5292-302issue
16eissn
0021-9193issn
1098-5530pii
186/16/5292journal_volume
186pub_type
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更新日期:1976-02-01 00:00:00
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doi:10.1128/JB.91.4.1618-1624.1966
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journal_title:Journal of bacteriology
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更新日期:1963-02-01 00:00:00
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journal_title:Journal of bacteriology
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pub_type: 杂志文章
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更新日期:1975-05-01 00:00:00
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pub_type: 杂志文章
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更新日期:2010-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2008-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.10.2811-2822.2000
更新日期:2000-05-01 00:00:00
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pub_type: 杂志文章
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更新日期:2005-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.129.2.1121-1128.1977
更新日期:1977-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.153.1.571-573.1983
更新日期:1983-01-01 00:00:00