Identification of the Novobiocin Biosynthetic Gene Cluster of
            <i>Streptomyces spheroides</i>
            NCIB 11891

Steffensky, Marion; Mühlenweg, Agnes; Wang, Zhaoxin; Li, Shu-Ming; Heide, Lutz

doi:10.1128/aac.44.5.1214-1222.2000

Cited by 187 publications

(179 citation statements)

References 60 publications

(45 reference statements)

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Section: Discovery Of a New Subgroup Of Aromatic Ptasesmentioning

confidence: 99%

“…In vitro biochemical characterization of the encoded gene product demonstrated that 4-hydroxyphenylpyruvate (4-HPP) served as the aromatic acceptor with the C5 isoprenoid diphosphate, DMAPP, serving as the prenyl donor in a divalent cationindependent reaction [14]. Moreover, the closely related NovQ protein found in S. spheroides [15] (84% sequence identity to CloQ) was speculated to participate in the dimethylallylation of Ring A of the structurally related aminocoumarin novobiocin [14].Two years after these initial discoveries, an open reading frame designated orf2 (now renamed nphB) was identified based onits similarityto cloQ/novQ in a gene cluster associated with the biosynthesis of the geranylated natural product naphterpin in Streptomyces sp. strain CL190 [16].…”

mentioning

confidence: 99%

See 1 more Smart Citation

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

Tello

Kuzuyama²,

Heide³

et al. 2008

Cell. Mol. Life Sci.

127

155

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Aromatic prenyltransferases (PTases) catalyze the transfer of a C5 (dimethylallyl), C10 (geranyl) or C15 (farnesyl) prenyl group derived from the corresponding isoprenyl diphosphate metabolites onto a variety of electron-rich aromatic acceptors. Prenyl groups appear in a wide variety of bioactive natural products of microbial and plant origin, including amino acids, stilbenes, alkaloids, polyketides and phenylpropanoids such as flavonoids, creating natural product hybrids with altered or enhanced bioactivities. Prenylation of flavonoids enhances some of the desirable pharmacological properties of these plant compounds [1], as demonstrated for apigenin and liquiritigenin [2]. Prenylation appears in many cases to provide a higher level of bioactivity compared to the non-prenylated precursor, often by increasing affinity for biological membranes and interactions with cellular targets [3]. With the recent identification of these enzymes there is increased interest in the role of these regiospecific catalysts in expanding the diversity and bioactivities of several important classes of natural products in vivo and in vitro.One way in which PTases can be categorized depends on whether they catalyze the synthesis of isoprenyl diphosphates, the prenylation of a protein or the prenylation of an aromatic substrate. Isoprenyl diphosphate synthases catalyze the chain elongation of an allylic isoprenyl diphosphate substrate by reaction with isopentenyl diphosphate [4]. Protein PTases transfer a geranyl-geranyl or farnesyl group to the Cys residue on a CaaX motif at the C-terminus of several proteins to facilitate membrane anchoring in eukaryotes and possibly archaea [5]. Small-molecule aromatic PTases constituting the third category can be subdivided into membrane-associated and functionally soluble PTases. Membrane-associated PTases contain a characteristic (N/D)DXXD Mg 2+ -diphosphate binding motif which is also found in the isoprenyl diphosphate synthases and are involved e.g. in the biosynthesis of ubiquinones and © Birkhäuser Verlag, Basel, 2008 Richard@salk.edu. [6], in the biosynthesis of membrane lipids in archaea [7] and in the formation of plant secondary metabolites [8].The functionally soluble PTases do not possess an obvious Mg 2+ -diphosphate binding motif. Moreover, most of the currently known soluble aromatic PTases segregate into two subgroups based on their primary sequence similarity and on the small-molecule substrates they accept for prenylation. The first subgroup commonly prenylates indole-containing ring systems. Examples of these PTases include the fungal enzymes FgaPT1 and FgaPT2 involved in the biosynthesis of fumigaclavin C [9], as well as the newly discovered N-reverse PTase CdpNPT [10] and the 7-dimethylallyl-tryptophan synthase Afu3 g12930 [11]. LtxC, which is involved in the biosynthesis of lyngbyatoxins obtained from the cyanobacteria Lyngbya majuscula [12], also prenylates an indole-containing ring system but shares no sequence similaritywith the fungal enzymes named above.The second subg...

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Section: Discovery Of a New Subgroup Of Aromatic Ptasesmentioning

confidence: 99%

mentioning

confidence: 99%

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

Tello

Kuzuyama²,

Heide³

et al. 2008

Cell. Mol. Life Sci.

127

155

View full text Add to dashboard Cite

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“…Biochemical characterization of the P450s that catalyze the oxidation of carrier protein-bound amino acids in novobiocin and nikkomycin biosynthesis has been performed (23,25). Walsh and co-workers demonstrated that the oxidation of tyrosine to ␤-R-hydroxytyrosine is catalyzed by the P450 NovI, with tyrosine bound to another protein in the novobiocin biosynthetic operon, NovH.…”

mentioning

confidence: 99%

Structural Characterization of OxyD, a Cytochrome P450 Involved in β-Hydroxytyrosine Formation in Vancomycin Biosynthesis

Cryle

Meinhart

Schlichting

2010

Journal of Biological Chemistry

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The cytochrome P450 OxyD from the balhimycin glycopeptide antibiotic biosynthetic operon of Amycolatopsis mediterranei is involved in the biosynthesis of the modified amino acid ␤-R-hydroxytyrosine, an essential precursor for biosynthesis of the vancomycin-type aglycone. OxyD binds the substrate tyrosine not free in solution, but rather covalently linked to the carrier protein (CP) domain of the non-ribosomal peptide synthase BpsD, exhibiting micromolar binding affinity to a tyrosineloaded carrier protein construct. The crystal structure of OxyD was determined to 2.1-Å resolution, revealing a potential binding site for the carrier protein-bound substrate in a different orientation to that seen with the acyl carrier protein-bound P450 BioI Hydroxylated amino acid residues are widely utilized in nature as precursors for complex natural products (see Fig. 1). The oxidative modification of amino acids at the ␤-position is a process that occurs in many different classes of medicinally important biomolecules, including the vancomycin-type antibiotics (reviewed in Refs. 2, 3). These glycopeptide antibiotics have had great clinical success in treating Gram-positive bacterial infections that are resistant to other classes of antibiotics and function through blocking cell wall biosynthesis via complex formation with the cell wall precursor peptidoglycan peptidyl units terminating in -Lys-D-Ala-D-Ala (2, 3). The biosynthesis of the vancomycin peptide requires the incorporation of two amino acid residues derived from ␤-R-hydroxytyrosine. Another group of medicinally important molecules that contain hydroxylated amino acid residues is the aminocoumarincontaining angucycline family of antibiotic compounds, which include coumermycin A, clorobiocin, and simocyclinone and act upon bacterial DNA gyrase. ␤-R-Hydroxytyrosine is an essential precursor in the biosynthesis of these types of antibiotics, where it is needed for the formation of both the A (Fig. 1, blue) and B (Fig. 1, red) rings in the aminocoumarin core. Further examples of the incorporation of hydroxylated amino acids into the biosyntheses of medicinally important compounds can be found in the biosyntheses of the nikkomycin-type antibiotics, zorbamycin and echinomycin ( Fig. 1) (4 -7).Despite the large structural difference in the compounds highlighted above and the difference in the biosyntheses of the compounds overall, the formation of the ␤-hydroxyamino acid precursors follows a conserved route. This involves the oxidation of the amino acid by a member of the cytochrome P450 (P450) 2 superfamily. P450s form a superfamily of heme containing monoxygenases that catalyze a vast array of biologically important transformations (8). The archetypical P450 reaction is the hydroxylation of unactivated C-H bonds, a difficult oxidation made more impressive by the ability of P450s to utilize their oxidative power both regio-and stereoselectively. Many P450s play important roles in natural product biosyntheses, where such selectivity is channeled toward the synthesis of comp...

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“…The proposed role for NovJ and NovK was first intuited from their location adjacent to noVHI in the noV gene cluster (noVH,I,J,K) (6) and from bioinformatics analysis that indicates that they belong to the nicotinamide-dependent -keto acid reductase superfamily (30). Most intriguingly, we noted homology to a pair of orfs, bbsC and bbsD, the predicted -oxidases in anaerobic toluene catabolism (31).…”

Section: Discussionmentioning

confidence: 73%

“…These studies are enabled by the DNA sequence determinations of the biosynthetic gene clusters for all three of the aminocoumarin antibiotics, novobiocin, clorobiocin, and coumermycin, by the Heide group in Tubingen (6)(7)(8).…”

mentioning

confidence: 99%

NovJ/NovK Catalyze Benzylic Oxidation of a β-Hydroxyl Tyrosyl-S-pantetheinyl Enzyme during Aminocoumarin Ring Formation in Novobiocin Biosynthesis

Pacholec

2005

Biochemistry

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The bicyclic coumarin ring in the aminocoumarin natural product antibiotics that target bacterial DNA gyrase is assembled from tyrosine by nonribosomal peptide synthetase logic. Tyrosine has previously been shown to be activated and installed as a phosphopantetheinyl thioester on the thiolation domain of NovH and then hydroxylated on the benzylic carbon by the heme protein NovI, generating -OH-Tyr-S-NovH. This aminoacyl-S-protein is the substrate for the next two orfs, Streptomyces sphaeroides NovJ and NovK, that have now been expressed in and purified from Escherichia coli as a J 2 K 2 heterotetramer. NovJ/NovK use NADP as an electron acceptor to oxidize the -OH of the tyrosyl moiety to yield the tethered -ketotyrosyl-S-NovH. The enol tautomer is the form that predominates in the subsequently cyclized aminocoumarin scaffold. The labile -ketotyrosyl thioester moiety was identified by hydrolytic release from NovH, analysis by mass spectroscopy, and comparison with a synthetic sample. We also have identified a residue in NovJ that when mutated results in a 50-fold reduction in catalytic activity.

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Identification of the Novobiocin Biosynthetic Gene Cluster of Streptomyces spheroides NCIB 11891

Cited by 187 publications

References 60 publications

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

Structural Characterization of OxyD, a Cytochrome P450 Involved in β-Hydroxytyrosine Formation in Vancomycin Biosynthesis

NovJ/NovK Catalyze Benzylic Oxidation of a β-Hydroxyl Tyrosyl-S-pantetheinyl Enzyme during Aminocoumarin Ring Formation in Novobiocin Biosynthesis

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