Isolation and Identification of Three New 5‐Alkenyl‐3,3(2H)‐furanones from Two Streptomyces Species Using a Genomic Screening Approach.

Banskota, Arjun H.; McAlpine, James B.; Soerensen, Dan; Aouidate, Mustapha; Piraee, Mahmood; Alarco, Anne‐Marie; Ōmura, Satoshi; Shiomi, Kazuro; Farnet, Chris M.; Zazopoulos, Emmanuel

doi:10.1002/chin.200639222

Cited by 6 publications

(12 citation statements)

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“…The column was subsequently eluted with a step gradient of EtOH/aqueous NH 4 HCO 3 buffer pH 10 to collect one 500 ml fraction and then seven fractions (200 ml each) i.e., 1 : 9 (fraction 1); 1 : 4 (fraction 2); 3 : 7 (fraction 3); 2 : 3 (fraction 4); 1 : 1 (fraction 5); 3 : 2 (fraction 6); 8 : 1 (fraction 7) and EtOH (fraction 8). Fractions 4ϳ7 were pooled, concentrated and the residue was subjected for HPLC (Waters Autopurification System with ACD), using a Symmetry C18 column (5 m , 30ϫ100 mm) with a gradient of 10 mM aqueous NH 4 OAc, (adjusted to pH 5 with glacial AcOH)/acetonitrile 74 : 26 v/v to 50 : 50 over 20 minutes at 39 ml/minute. The collection was triggered by UV absorption at 261 nm (PDA).…”

Section: Methods Bmentioning

confidence: 99%

“…The DNA and protein sequences that comprise the ECO-0501 gene cluster are deposited in GeneBank under accession numbers Contig1 DQ884174, orientalis ATCC 43491 was grown on ISP2 agar (Difco) for 5 to 7 days, and the surface growth from the agar plate was homogenized and transferred to a 125 ml flask containing three glass beads (5 mm diameter), and 25 ml of sterile medium prepared from trypticase soy broth (Bacto) 30 g, yeast extract 3 g, MgSO 4 2 g, glucose 5 g, maltose 4 g, to which one liter distilled water was added. This vegetative culture was incubated at 28°C for about 60 hours on a shaker with a 2.5 cm throw and set at 250 rpm.…”

Section: Genome Scanningmentioning

confidence: 99%

“…ECO-0501 (1, 37.4 mg/liter) was purified by repeated HPLC on a Waters Autopurification System with ACD using a Waters RCM Column (Novapak C-18, 6 m , 40ϫ200 mm) with a gradient of 10 mM aqueous NH 4 OAc adjusted to pH 5 with glacial AcOH/acetonitrile from 80 : 20 v/v to 20 : 80 over 25 minutes at 35 ml/minute.…”

Section: Isolation Of Eco-0501 (1)mentioning

confidence: 99%

“…This approach not only ascertains the potential of a producing organism, but it provides a handle to detect, isolate and structurally define a specific metabolite. We have demonstrated this approach in the isolation and structural determination of an antifungal Part 3 † ECO-0501, a Novel Antibacterial of a New Class agent, ECO-02301 from Streptomyces aizunensis [3] and three 5-alkenyl-3,3(2H)-furanones from two different Streptomyces species [4]. In this article, we are describing the use of the genome scanning technique [5,6] to identify and isolate a novel class of antibacterial (ECO-0501) from the Amycolatopsis orientalis ATCC 43491 strain, which was deposited as a vancomycin producer.…”

Section: Introductionmentioning

confidence: 99%

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Genomic Analyses Lead to Novel Secondary Metabolites

et al. 2006

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Genomic analyses of Amycolatopsis orientalis ATCC 43491 strain, deposited as a vancomycin producer, revealed the presence of genetic loci for the production of at least 10 secondary metabolites other than vancomycin. One of these gene clusters, which contained a type I polyketide synthase, was predicted to direct the synthesis of novel class of compound, a glycosidic polyketide ECO-0501 (1). Screening of culture extracts for a compound with the predicted physicochemical properties of the product from this locus, led to the isolation of the 13-Oglucuronide of 13-hydroxy-2,12,14,16,22-pentamethyl-28-(N-methyl-guanidino)-octacosa-2,4,6,8,10,14,20,24-octaenoic acid (2-hydroxy-5-oxo-cyclopent-1-enyl)-amide (ECO-0501, 1). The structure, confirmed by spectral analyses including MS, and 1D and 2D NMR experiments, were in accord with that predicted by genomic analyses. ECO-0501 possessed strong antibacterial activity against a series of Gram-positive pathogens including several strains of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE). ECO-0501 was chemically modified by esterification (1aϳ1c), Nacetylation (1d) and hydrogenation (1e) in order to explore structure activity relationships (SAR). Keywords Amycolatopsis orientalis, ECO-0501, antibacterial, PKS I IntroductionDrug-resistant bacterial infections are a growing health concern. Resistance has been developed to every major class of antibiotics on the market, and an increasing number of pathogenic bacteria are becoming resistant to multiple classes of antibiotics, thereby limiting treatment options. Hence, there is a renewed urgency for the discovery of new classes of antibiotics for the treatment of drug resistant bacterial infections. To accelerate the discovery of such potential antibacterial candidates from natural resources a new, fast and efficient technology is needed. The genomics of secondary metabolite biosynthesis recently evolved to the point where analysis of the genome of an organism can define its biosynthetic capabilities for secondary metabolites. A genome scanning technique that has been developed in our laboratories, and used with our DECIPHER ® technology to analyze the genomes of actinomycetes for their secondary metabolite biosynthetic genes, greatly reduces the amount of sequencing required to define this capability [1,2]. This approach not only ascertains the potential of a producing organism, but it provides a handle to detect, isolate and structurally define a specific metabolite. We have demonstrated this approach in the isolation and structural determination of an antifungal Genomic Analyses Lead to Novel Secondary MetabolitesPart 3

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Section: Methods Bmentioning

confidence: 99%

Section: Genome Scanningmentioning

confidence: 99%

Section: Isolation Of Eco-0501 (1)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Genomic Analyses Lead to Novel Secondary Metabolites

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“…Thus, these Streptomyces seem to produce hitherto unknown polyketides that had not been identified by traditional activity-based screenings 8,9) . Since the presence of novel pks genes in an organism is a good indicator of production of novel polyketide molecules in this organism, the sequencing of pks genes is often used as a screening method for the discovery of novel polyketides [10][11][12][13][14][15] . For example, analysis of novel pks genes in a rubromycin-producer led to the discovery of two novel angucyclines 16) .…”

Section: Introductionmentioning

confidence: 99%

Sequence Diversity of Type-II Polyketide Synthase Genes in Streptomyces

Komaki

Harayama

2006

Actinomycetologica

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Type-II polyketide synthases (PKS) are composed of two b-ketoacyl synthase subunits (KSa and KSb) and one acyl carrier protein, which are involved in the biosynthesis of polyketide molecules. In this study, we investigated the diversity of type-II pks genes in 83 Streptomyces strains listed in the catalogue of the NITE Biological Resource Center (NBRC, http://www.nbrc.nite.go.jp/e/index.html) by determining partial sequences of the KSa genes. As expected, KSa genes were detected in 9 strains that have been reported to produce aromatic-polyketides. More interestingly, novel KSa genes were detected in 55 out of 74 Streptomyces strains so far unknown to produce aromatic antibiotics. The antibiotic-type KSa genes exhibited a high degree of divergence, the pairwise nucleotide similarities ranged from 67 to 95%. The existence of novel cryptic KSa genes in the Streptomyces strains suggest their potential to synthesize new aromatic polyketides.

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Novel approaches to discovery of antibacterial agents

Taylor

Wright

2008

Anim. Health. Res. Rev.

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Antimicrobial resistance is a rapidly increasing problem impacting the successful treatment of bacterial infectious disease. To combat resistance, the development of new treatment options is required. Recent advances in technology have aided in the discovery of novel antibacterial agents, specifically through genome mining for novel natural product biosynthetic gene clusters and improved small molecule high-throughput screening methods. Novel targets such as lipopolysaccharide and fatty acid biosyntheses have been identified by essential gene studies, representing a shift from traditional antibiotic targets. Finally, inhibiting non-essential genes with small molecules is being explored as a method for rescuing the activity of 'old' antibiotics, providing a novel synergistic approach to antimicrobial discovery.

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Isolation and Identification of Three New 5‐Alkenyl‐3,3(2H)‐furanones from Two Streptomyces Species Using a Genomic Screening Approach.

Cited by 6 publications

References 1 publication

Genomic Analyses Lead to Novel Secondary Metabolites

Genomic Analyses Lead to Novel Secondary Metabolites

Sequence Diversity of Type-II Polyketide Synthase Genes in Streptomyces

Novel approaches to discovery of antibacterial agents

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