Molecular cloning and sequence analysis of the clorobiocin biosynthetic gene cluster: new insights into the biosynthesis of aminocoumarin antibiotics
               The GenBank accession number for the sequence of cosmid K1F2 is AF329398.

Pojer, F.; Li, Shu-Ming; Heide, Lutz

doi:10.1099/00221287-148-12-3901

Cited by 138 publications

(134 citation statements)

References 34 publications

(37 reference statements)

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“…It was identified in the gene cluster involved in the biosynthesis of Ring A (3-dimethylallyl-4-hydroxybenzoic acid) of clorobiocin, an aminocoumarin antibiotic produced by Streptomyces roseochromogenes [13]. 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].…”

Section: Discovery Of a New Subgroup Of Aromatic Ptasesmentioning

confidence: 99%

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

Tello

Kuzuyama²,

Heide³

et al. 2008

Cell. Mol. Life Sci.

128

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%

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

Tello

Kuzuyama²,

Heide³

et al. 2008

Cell. Mol. Life Sci.

128

155

View full text Add to dashboard Cite

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“…The composition of the media and the culture conditions used for S. roseochromogenes var. oscitans DS 12.976 (Aventis) were as described previously (Pojer et al, 2002). For isolation of chromosomal DNA and preparation of protoplasts, S. roseochromogenes was grown in CRM medium containing 0?75 % glycine.…”

Section: Methodsmentioning

confidence: 99%

“…oscitans DS 12.976 (Pojer et al, 2002). A comparison of these gene clusters showed that the structural differences and similarities between the three antibiotics are reflected perfectly by the differences and similarities in the organization of the corresponding biosynthetic gene clusters (Pojer et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

“…The novobiocin gene cluster contains a carbamoyltransferase gene, novN. At the same relative position, a group of seven genes, cloN1 to cloN7 and the homologous couN1 to couN7, are found in the gene clusters of clorobiocin and coumermycin A 1 , respectively (Pojer et al, 2002). Our earlier study (Xu et al, 2002) showed that couN3 and couN4 are involved in the biosynthesis of the pyrrole-2-carboxylic acid in coumermycin A 1 .…”

Section: Introductionmentioning

confidence: 99%

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CloN2, a novel acyltransferase involved in the attachment of the pyrrole-2-carboxyl moiety to the deoxysugar of clorobiocin

Kahlich

Kammerer

et al. 2003

Microbiology

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The aminocoumarin antibiotic clorobiocin contains a 5-methylpyrrole-2-carboxylic acid unit, attached via an ester bond to the 3-OH group of the deoxysugar moiety. To investigate candidate genes responsible for the formation of this ester bond, a gene inactivation experiment was carried out in the clorobiocin producer Streptomyces roseochromogenes var. oscitans DS 12.976. An in-frame deletion was created in the coding sequence of the gene cloN2. The production of secondary metabolites in the wild-type and in the cloN2 mutant was analysed. The wild-type showed clorobiocin as the main product, whereas the cloN2 mutant accumulated a new aminocoumarin derivative, novclobiocin 104, lacking the pyrrole moiety at the 3-OH of the deoxysugar. In addition, free pyrrole-2-carboxylic acid accumulated in the culture extract of the cloN2 mutant. The structures of the metabolites were confirmed by NMR and LC-MS analysis. Clorobiocin production was successfully restored in the cloN2 mutant by introducing a replicative plasmid containing the cloN2 sequence. These results prove an involvement of cloN2 in the formation of the ester bond between the pyrrole moiety and the deoxysugar in clorobiocin biosynthesis. Furthermore, they indicate that the C-methylation at position 5 of the pyrrole moiety occurs after the attachment of pyrrole-2-carboxylic acid unit to the deoxysugar moiety.

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“…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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Molecular cloning and sequence analysis of the clorobiocin biosynthetic gene cluster: new insights into the biosynthesis of aminocoumarin antibiotics The GenBank accession number for the sequence of cosmid K1F2 is AF329398.

Cited by 138 publications

References 34 publications

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

The ABBA family of aromatic prenyltransferases: broadening natural product diversity

CloN2, a novel acyltransferase involved in the attachment of the pyrrole-2-carboxyl moiety to the deoxysugar of clorobiocin

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

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