Cloning and sequence analysis of the highly expressed melanin-synthesizing gene operon from Streptomyces castaneoglobisporus

Ikeda, Kayo; Masujima, Tsutomu; Suzuki, Ken; Sugiyama, Masanori

doi:10.1007/s002530050652

Cited by 42 publications

(33 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2 The gene cluster contains 13 open reading frames. Of the 13 gene products, two proteins (GriE and GriF) are similar to the MelC1 and MelC2 proteins, respectively, both of which are responsible for melanin biosynthesis in streptomycetes (17)(18)(19)(20)(21). MelC2 tyrosinase belongs to the type 3 copper protein family, and MelC1 serves as a copper chaperon for MelC2.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

A Novel o-Aminophenol Oxidase Responsible for Formation of the Phenoxazinone Chromophore of Grixazone

Suzuki

Furusho

Higashi

et al. 2006

Journal of Biological Chemistry

View full text Add to dashboard Cite

Grixazone contains a phenoxazinone chromophore and is a secondary metabolite produced by Streptomyces griseus. In the grixazone biosynthesis gene cluster, griF (encoding a tyrosinase homolog) and griE (encoding a protein similar to copper chaperons for tyrosinases) are encoded. An expression study of GriE and GriF in Escherichia coli showed that GriE activated GriF by transferring copper ions to GriF, as has been observed for a Streptomyces melanogenesis system in which the MelC1 copper chaperon transfers copper ions to MelC2 tyrosinase. In contrast with tyrosinases, GriF showed no monophenolase activity, although it oxidized various o-aminophenols as preferable substrates rather than catechol-type substrates. Deletion of the griEF locus on the chromosome resulted in accumulation of 3-amino-4-hydroxybenzaldehyde (3,4-AHBAL) and its acetylated compound, 3-acetylamino-4-hydroxybenzaldehyde. GriF oxidized 3,4-AHBAL to yield an o-quinone imine derivative, which was then non-enzymatically coupled with another molecule of the o-quinone imine to form a phenoxazinone. The coexistence of N-acetylcysteine in the in vitro oxidation of 3,4-AH-BAL by GriF resulted in the formation of grixazone A, suggesting that the -SH group of N-acetylcysteine is conjugated to the o-quinone imine formed from 3,4-AHBAL and that the conjugate is presumably coupled with another molecule of the o-quinone imine. GriF is thus a novel o-aminophenol oxidase that is responsible for the formation of the phenoxazinone chromophore in the grixazone biosynthetic pathway.We have long studied the A-factor regulatory cascade that leads to secondary metabolite formation and morphological differentiation in Streptomyces griseus (1, 2). The A-factor (2-isocapryloyl-3R-hydroxymethyl-␥-butyrolactone) triggers the synthesis of almost all the secondary metabolites produced by this species. One of the secondary metabolites under the control of the A-factor is grixazone. Grixazone is a yellow pigment and actually a mixture of grixazones A and B (compounds 1a and 1b) (see Fig. 2C) (3). Grixazone A is a novel compound, and grixazone B has been reported to show a parasiticide activity (4).Grixazones contain a phenoxazinone chromophore. The phenoxazinone skeleton is common to actinomycin D produced by Streptomyces antibioticus (5), michigazone produced by Streptomyces michiganensis (6), texazone produced by Streptomyces sp. WRAT-210 (7), exfoliazone produced by Streptomyces exfoliatus (8), and 4-demethoxymichigazone produced by Streptomyces halstedii (9). Hsieh and Jones (10) reported a phenoxazinone synthase in S. antibioticus that catalyzes the six-electron oxidative coupling of o-aminophenol compounds derived from tryptophan through 3-hydroxyanthranilic acid. However, disruption of the phenoxazinone synthase gene in S. antibioticus does not affect actinomycin D synthesis, showing that the phenoxazinone skeleton in actinomycin D is biosynthesized in vivo by a still unknown enzyme or non-enzymatically (11). On the other hand, michigazone with a hydroxymethyl group at ...

show abstract

Section: Resultsmentioning

confidence: 99%

“…The amino acid sequences of MelC1 in streptomycetes are conserved at a moderate level (40 -50% identity) and are always encoded as a neighbor of melC2 (17,20). GriE was therefore expected to be involved in the transfer of copper ion to the apo form of GriF via binary complex formation (23,24).…”

Section: Resultsmentioning

confidence: 99%

A Novel o-Aminophenol Oxidase Responsible for Formation of the Phenoxazinone Chromophore of Grixazone

Suzuki

Furusho

Higashi

et al. 2006

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…coelicolor may produce molecules whose structures are related to the structures of adrenaline pathway intermediates that could serve as substrates for the SCO2525 methyltransferase. Streptomyces species typically are able to synthesize DOPA from tyrosine, and the DOPA is then utilized in melanin pigment biosynthesis (17). While S. coelicolor does not make melanin, its genome does contain the melC2 gene, whose product is a monophenol monooxygenase (tyrosinase) that should be capable of catalyzing DOPA production (35).…”

Section: Discussionmentioning

confidence: 99%

Novel Genes That Influence Development inStreptomyces coelicolor

et al. 2004

View full text Add to dashboard Cite

Filamentous soil bacteria of the genus Streptomyces carry out complex developmental cycles that result in sporulation and production of numerous secondary metabolites with pharmaceutically important activities. To further characterize the molecular basis of these developmental events, we screened for mutants of Streptomyces coelicolor that exhibit aberrant morphological differentiation and/or secondary metabolite production. On the basis of this screening analysis and the subsequent complementation analysis of the mutants obtained we assigned developmental roles to a gene involved in methionine biosynthesis (metH) and two previously uncharacterized genes (SCO6938 and SCO2525) and we reidentified two previously described developmental genes (bldA and bldM). In contrast to most previously studied genes involved in development, the genes newly identified in the present study all appear to encode biosynthetic enzymes instead of regulatory proteins. The MetH methionine synthase appears to be required for conversion of aerial hyphae into chains of spores, SCO6938 is a probable acyl coenzyme A dehydrogenase that contributes to the proper timing of aerial mycelium formation and antibiotic production, and SCO2525 is a putative methyltransferase that influences various aspects of colony growth and development.

show abstract

“…Streptomyces strains were grown at 28°C on an FB agar medium (9). To select objective clones, appropriate antibiotics were added to the medium (14).…”

Section: Methodsmentioning

confidence: 99%

Molecular Cloning and Heterologous Expression of a Biosynthetic Gene Cluster for the Antitubercular Agent d -Cycloserine Produced by Streptomyces lavendulae

Kumagai

Koyama

Oda

et al. 2010

Antimicrob Agents Chemother

View full text Add to dashboard Cite

In the present study, we successfully cloned a 21-kb DNA fragment containing a D-cycloserine (DCS) biosynthetic gene cluster from a DCS-producing Streptomyces lavendulae strain, ATCC 11924. The putative gene cluster consists of 10 open reading frames (ORFs), designated dcsA to dcsJ. This cluster includes two ORFs encoding D-alanyl-D-alanine ligase (dcsI) and a putative membrane protein (dcsJ) as the self-resistance determinants of the producer organism, indicated by our previous work. When the 10 ORFs were introduced into DCS-nonproducing Streptomyces lividans 66 as a heterologous host cell, the transformant acquired DCS productivity. This reveals that the introduced genes are responsible for the biosynthesis of DCS. As anticipated, the disruption of dcsG, seen in the DCS biosynthetic gene cluster, made it possible for the strain ATCC 11924 to lose its DCS production. We here propose the DCS biosynthetic pathway. The soil-dwelling genus Streptomyces undergoes a complex morphological differentiation and produces an enormous variety of bioactive secondary metabolites. Because they include clinically useful antibiotics and immunosuppressants, the genus Streptomyces occupies an important position as an industrial microorganism.D-Cycloserine (DCS), a cyclic structural analogue of D-alanine, is produced by "Streptomyces garyphalus" and Streptomyces lavendulae. This antibiotic is used as an antitubercular agent (21). Since DCS is similar to D-alanine, it prevents the action of both alanine racemase and D-alanyl-D-alanine ligase, which are necessary for the biosynthesis of a bacterial cell wall. Thus, DCS functions as an inhibitor of bacterial cell wall biosynthesis (16,19). Although the structure of DCS is very simple, the biosynthetic genes for DCS have never been cloned until now.In general, antibiotic biosynthetic genes form a cluster and are adjacent to their self-resistance genes. We have previously cloned a gene (orfB) that confers resistance to DCS on Streptomyces lividans and Escherichia coli from DCS-producing S. garyphalus (CSH) 5-12 (17). The sequence analysis suggests that the gene may encode a membrane protein that is necessary for the excretion of the DCS outside the cell. We have found that the same gene is also present in S. lavendulae ATCC 25233 (20). We have also previously demonstrated that Dalanyl-D-alanine ligase, which is a target enzyme of DCS, functions as a self-resistance determinant in S. lavendulae ATCC 25233 (20). Interestingly, the gene encoding D-alanyl-D-alanine ligase, designated ddlS, was located just upstream of the putative membrane protein gene, orfB (20). Although the selfresistance genes in the DCS producers have been thoroughly analyzed, no attempt to clone the DCS biosynthetic genes has been carried out yet.To clone the biosynthetic genes for DCS from another DCSproducing S. lavendulae strain, ATCC 11924, we first investigated whether this strain harbors orfB and ddlS. Since both genes were found to be conserved, the flanking region was cloned from the chromosomal DNA. Here, we ...

show abstract

Cloning and sequence analysis of the highly expressed melanin-synthesizing gene operon from Streptomyces castaneoglobisporus

Cited by 42 publications

References 15 publications

A Novel o-Aminophenol Oxidase Responsible for Formation of the Phenoxazinone Chromophore of Grixazone

A Novel o-Aminophenol Oxidase Responsible for Formation of the Phenoxazinone Chromophore of Grixazone

Novel Genes That Influence Development inStreptomyces coelicolor

Molecular Cloning and Heterologous Expression of a Biosynthetic Gene Cluster for the Antitubercular Agent d -Cycloserine Produced by Streptomyces lavendulae

Contact Info

Product

Resources

About