ArticleJournal of bacteriology2002
Expression, purification, and characterization of AknX anthrone oxygenase, which is involved in aklavinone biosynthesis in Streptomyces galilaeus.
Article in Journal of bacteriology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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20 citing papers in PubMed, 68 citations in OpenAlex.
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- Omics Technologies to Understand Activation of a Biosynthetic Gene Cluster in Micromonospora sp. WMMB235: Deciphering Keyicin Biosynthesis.ACS chemical biology · 2019Article
- The Enzymology of Organic Transformations: A Survey of Name Reactions in Biological Systems.Angewandte Chemie (International ed. in English) · 2017Review
- Monooxygenase Substrates Mimic Flavin to Catalyze Cofactorless Oxygenations.The Journal of biological chemistry · 2016Article
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- Baeyer-Villiger C-C bond cleavage reaction in gilvocarcin and jadomycin biosynthesis.Journal of the American Chemical Society · 2012Article
- F420H2-dependent degradation of aflatoxin and other furanocoumarins is widespread throughout the actinomycetales.PloS one · 2012Article
- Identification and characterization of two families of F420 H2-dependent reductases from Mycobacteria that catalyse aflatoxin degradation.Molecular microbiology · 2010Article
- Oxytetracycline biosynthesis.The Journal of biological chemistry · 2010Review
- HypC, the anthrone oxidase involved in aflatoxin biosynthesis.Applied and environmental microbiology · 2010Article
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- Multi-oxygenase complexes of the gilvocarcin and jadomycin biosyntheses.Journal of the American Chemical Society · 2007Article
- Insights in the glycosylation steps during biosynthesis of the antitumor anthracycline cosmomycin: characterization of two glycosyltransferase genes.Applied microbiology and biotechnology · 2006Article
- Premithramycinone G, an early shunt product of the mithramycin biosynthetic pathway accumulated upon inactivation of oxygenase MtmOII.Angewandte Chemie (International ed. in English) · 2006Article
- Engineered biosynthesis of a novel amidated polyketide, using the malonamyl-specific initiation module from the oxytetracycline polyketide synthase.Applied and environmental microbiology · 2006Article
- The structure of ActVA-Orf6, a novel type of monooxygenase involved in actinorhodin biosynthesis.The EMBO journal · 2003Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In streptomycete anthracycline biosynthetic gene clusters, small open reading frames are located just upstream of minimal polyketide synthase genes. aknX is such a gene found in the aklavinone-aclacinomycin biosynthetic gene cluster of Streptomyces galilaeus. In order to identify its function, the aknX gene was expressed in Escherichia coli. The cell extract prepared from E. coli cells overexpressing AknX protein exhibited anthrone oxygenase activity, which converted emodinanthrone to anthraquinone emodin. This indicates that AknX and related gene products such as DnrG and SnoaB are involved in the formation of aklanonic acid from its anthrone precursor, as suggested by their homology with TcmH and ActVA6. The AknX protein fused with a His(6) tag was efficiently purified to homogeneity by Ni(2+) affinity and anion-exchange column chromatography. The native molecular mass of AknX was estimated to be 42 kDa by gel filtration. Thus, native AknX is considered to have a homotrimeric subunit structure. AknX, like TcmH and ActVA6, possesses no apparent prosthetic group for oxygen activation. Site-directed mutagenesis was carried out to identify the key amino acid residue(s) involved in the oxygenation reaction. Of seven AknX mutants expressed, the W67F mutant showed significantly reduced oxygenase activity, suggesting the important role of the W67 residue in the AknX reaction. A possible mechanism for the reaction via peroxy anion intermediate is proposed.
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