scholarly journals High-Throughput Screening for Streptomyces Antibiotic Biosynthesis Activators

2012 ◽  
Vol 78 (12) ◽  
pp. 4526-4528 ◽  
Author(s):  
Li Chen ◽  
Yemin Wang ◽  
Hang Guo ◽  
Min Xu ◽  
Zixin Deng ◽  
...  
Keyword(s):  
Gene Cluster ◽  
High Throughput ◽  
High Throughput Screening ◽  
Streptomyces Clavuligerus ◽  
Cosmid Library ◽  
Screening Procedure ◽  
Antibiotic Biosynthesis ◽  
Biosynthesis Gene Cluster ◽  
Content Type ◽  
Biosynthesis Gene

ABSTRACTA genomic cosmid library ofStreptomyces clavuligeruswas constructed and transferred efficiently by conjugation toStreptomyces lividans, and 12 distinct groups of overlapping cosmid clones that activated the silent actinorhodin biosynthesis gene cluster were identified. This generally applicable high-throughput screening procedure greatly facilitates the identification of antibiotic biosynthesis activators.

scholarly journals Characterization of the Amicetin Biosynthesis Gene Cluster from Streptomyces vinaceusdrappus NRRL 2363 Implicates Two Alternative Strategies for Amide Bond Formation

2012 ◽  
Vol 78 (7) ◽  
pp. 2393-2401 ◽  
Author(s):  
Gaiyun Zhang ◽  
Haibo Zhang ◽  
Sumei Li ◽  
Ji Xiao ◽  
Guangtao Zhang ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Acyl Carrier Protein ◽  
Bond Formation ◽  
Antiviral Agent ◽  
Amide Bond ◽  
Biosynthesis Gene Cluster ◽  
Alternative Strategies ◽  
Content Type ◽  
Amide Bond Formation ◽  
Biosynthesis Gene

ABSTRACTAmicetin, an antibacterial and antiviral agent, belongs to a group of disaccharide nucleoside antibiotics featuring an α-(1→4)-glycoside bond in the disaccharide moiety. In this study, the amicetin biosynthesis gene cluster was cloned fromStreptomyces vinaceusdrappusNRRL 2363 and localized on a 37-kb contiguous DNA region. Heterologous expression of the amicetin biosynthesis gene cluster inStreptomyces lividansTK64 resulted in the production of amicetin and its analogues, thereby confirming the identity of theamigene cluster.In silicosequence analysis revealed that 21 genes were putatively involved in amicetin biosynthesis, including 3 for regulation and transportation, 10 for disaccharide biosynthesis, and 8 for the formation of the amicetin skeleton by the linkage of cytosine,p-aminobenzoic acid (PABA), and the terminal (+)-α-methylserine moieties. The inactivation of the benzoate coenzyme A (benzoate-CoA) ligase geneamiLand theN-acetyltransferase geneamiFled to two mutants that accumulated the same two compounds, cytosamine and 4-acetamido-3-hydroxybenzoic acid. These data indicated that AmiF functioned as an amide synthethase to link cytosine and PABA. The inactivation ofamiR, encoding an acyl-CoA-acyl carrier protein transacylase, resulted in the production of plicacetin and norplicacetin, indicating AmiR to be responsible for attachment of the terminal methylserine moiety to form another amide bond. These findings implicated two alternative strategies for amide bond formation in amicetin biosynthesis.

scholarly journals Engineering Anthracycline Biosynthesis toward Angucyclines

2003 ◽  
Vol 47 (4) ◽  
pp. 1291-1296 ◽  
Author(s):  
Mikko Metsä-Ketelä ◽  
Kaisa Palmu ◽  
Tero Kunnari ◽  
Kristiina Ylihonko ◽  
Pekka Mäntsälä
Keyword(s):  
Gene Cluster ◽  
Polyketide Synthase ◽  
Gene Cassette ◽  
Side Chain ◽  
Antibiotic Biosynthesis ◽  
Biosynthesis Gene Cluster ◽  
Biosynthesis Gene ◽  
Starter Unit

ABSTRACT The biosynthesis pathways of two anthracyclines, nogalamycin and aclacinomycin, were directed toward angucyclines by using an angucycline-specific cyclase, pgaF, isolated from a silent antibiotic biosynthesis gene cluster. Addition of pgaF to a gene cassette that harbored the early biosynthesis genes of nogalamycin resulted in the production of two known angucyclinone metabolites, rabelomycin and its precursor, UWM6. Substrate flexibility of pgaF was demonstrated by replacement of the nogalamycin minimal polyketide synthase genes in the gene cassette with the equivalent aclacinomycin genes together with aknE2 and aknF, which specify the unusual propionate starter unit in aclacinomycin biosynthesis. This modification led to the production of a novel angucyclinone, MM2002, in which the expected ethyl side chain was incorporated into the fourth ring.

scholarly journals Discovery of a Widely Distributed Antibiotic Biosynthesis Gene Cluster

2008 ◽  
Vol 22 (S1) ◽  
Author(s):  
Douglas A Mitchell ◽  
Shaun Lee ◽  
Andrew L. Markley ◽  
Mary E. Hensler ◽  
Jack E. Dixon
Keyword(s):  
Gene Cluster ◽  
Antibiotic Biosynthesis ◽  
Biosynthesis Gene Cluster ◽  
Biosynthesis Gene

scholarly journals Whole-Genome Sequencing of the Fungus Penicillium citrinum Reveals the Biosynthesis Gene Cluster for the Mycotoxin Citrinin

2019 ◽  
Vol 8 (4) ◽  
Author(s):  
Markus Schmidt-Heydt ◽  
Dominic Stoll ◽  
Rolf Geisen
Keyword(s):  
Antibiotic Resistance ◽  
Whole Genome Sequencing ◽  
Gene Cluster ◽  
Genome Sequencing ◽  
Penicillium Citrinum ◽  
Whole Genome ◽  
Biosynthesis Gene Cluster ◽  
Content Type ◽  
Biosynthesis Gene

Penicillium citrinum is a food-contaminating ascomycete that consistently produces large amounts of the mycotoxin citrinin. Citrinin exhibits, besides its toxicity, antibiotic effects and thus potentially forces antibiotic resistance.

scholarly journals Genes for Production of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus Are Clustered with the cagA Gene That Encodes the C-1027 Apoprotein

2000 ◽  
Vol 44 (2) ◽  
pp. 382-392 ◽  
Author(s):  
Wen Liu ◽  
Ben Shen
Keyword(s):  
Sequence Analysis ◽  
Gene Cluster ◽  
Antibiotic Biosynthesis ◽  
Antitumor Antibiotic ◽  
Caga Gene ◽  
Biosynthesis Gene Cluster ◽  
Starting Point ◽  
Biosynthesis Gene ◽  
Cloned Gene

ABSTRACT C-1027, the most potent member of the enediyne antitumor antibiotic family, is produced by Streptomyces globisporus C-1027 and consists of an apoprotein (encoded by the cagA gene) and a nonpeptidic chromophore. The C-1027 chromophore could be viewed as being derived biosynthetically from a benzoxazolinate, a deoxyamino hexose, a β-amino acid, and an enediyne core. By adopting a strategy for cloning of the C-1027 biosynthesis gene cluster by mapping a putative dNDP-glucose 4,6-dehydratase (NGDH) gene to cagA, we have localized 75 kb of contiguous DNA from S. globisporus. DNA sequence analysis of two regions of the cloned gene cluster revealed two genes, sgcA and sgcB, that encode an NGDH enzyme and a transmembrane efflux protein, respectively, and confirmed that the cagA gene resides approximately 14 kb upstream of the sgcAB locus. The involvement of the cloned gene cluster in C-1027 biosynthesis was demonstrated by disrupting the sgcA gene to generate C-1027-nonproducing mutants and by complementing the sgcAmutants in vivo to restore C-1027 production. These results represent the first cloning of a gene cluster for enediyne antitumor antibiotic biosynthesis and provide a starting point for future genetic and biochemical investigations of C-1027 biosynthesis.

scholarly journals The Swinholide Biosynthesis Gene Cluster from a Terrestrial Cyanobacterium, Nostoc sp. Strain UHCC 0450

2017 ◽  
Vol 84 (3) ◽  
Author(s):  
Anu Humisto ◽  
Jouni Jokela ◽  
Liwei Liu ◽  
Matti Wahlsten ◽  
Hao Wang ◽  
...  
Keyword(s):  
Natural Products ◽  
Actin Cytoskeleton ◽  
Gene Cluster ◽  
Marine Sponge ◽  
Gene Clusters ◽  
Fold Axis ◽  
Biosynthesis Gene Cluster ◽  
Content Type ◽  
Biosynthesis Gene ◽  
Axis Of Symmetry

ABSTRACT Swinholides are 42-carbon ring polyketides with a 2-fold axis of symmetry. They are potent cytotoxins that disrupt the actin cytoskeleton. Swinholides were discovered from the marine sponge Theonella sp. and were long suspected to be produced by symbiotic bacteria. Misakinolide, a structural variant of swinholide, was recently demonstrated to be the product of a symbiotic heterotrophic proteobacterium. Here, we report the production of swinholide A by an axenic strain of the terrestrial cyanobacterium Nostoc sp. strain UHCC 0450. We located the 85-kb trans -AT polyketide synthase (PKS) swinholide biosynthesis gene cluster from a draft genome of Nostoc sp. UHCC 0450. The swinholide and misakinolide biosynthesis gene clusters share an almost identical order of catalytic domains, with 85% nucleotide sequence identity, and they group together in phylogenetic analysis. Our results resolve speculation around the true producer of swinholides and demonstrate that bacteria belonging to two distantly related phyla both produce structural variants of the same natural product. In addition, we described a biosynthesis cluster from Anabaena sp. strain UHCC 0451 for the synthesis of the cytotoxic and antifungal scytophycin. All of these biosynthesis gene clusters were closely related to each other and created a group of cytotoxic macrolide compounds produced by trans -AT PKSs of cyanobacteria and proteobacteria. IMPORTANCE Many of the drugs in use today originate from natural products. New candidate compounds for drug development are needed due to increased drug resistance. An increased knowledge of the biosynthesis of bioactive compounds can be used to aid chemical synthesis to produce novel drugs. Here, we show that a terrestrial axenic culture of Nostoc cyanobacterium produces swinholides, which have been previously found only from marine sponge or samples related to them. Swinholides are polyketides with a 2-fold axis of symmetry, and they are potent cytotoxins that disrupt the actin cytoskeleton. We describe the biosynthesis gene clusters of swinholide from Nostoc cyanobacteria, as well as the related cytotoxic and antifungal scytophycin from Anabaena cyanobacteria, and we study the evolution of their trans -AT polyketide synthases. Interestingly, swinholide is closely related to misakinolide produced by a symbiotic heterotrophic proteobacterium, demonstrating that bacteria belonging to two distantly related phyla and different habitats can produce similar natural products.

scholarly journals Identification of a Polyketide Synthase Gene Responsible for Ascochitine Biosynthesis in Ascochyta fabae and Its Abrogation in Sister Taxa

mSphere ◽  
2019 ◽  
Vol 4 (5) ◽  
Author(s):  
Wonyong Kim ◽  
Judith Lichtenzveig ◽  
Robert A. Syme ◽  
Angela H. Williams ◽  
Tobin L. Peever ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Secondary Metabolite ◽  
Polyketide Synthase ◽  
Animal Health ◽  
Gene Clusters ◽  
Biosynthesis Gene Cluster ◽  
Content Type ◽  
Pathogenicity Factor ◽  
Biosynthesis Gene ◽  
Ascochyta Fabae

ABSTRACT The polyketide-derived secondary metabolite ascochitine is produced by species in the Didymellaceae family, including but not restricted to Ascochyta species pathogens of cool-season food legumes. Ascochitine is structurally similar to the well-known mycotoxin citrinin and exhibits broad-spectrum phytotoxicity and antimicrobial activities. Here, we identified a polyketide synthase (PKS) gene (denoted pksAC) responsible for ascochitine production in the filamentous fungus Ascochyta fabae. Deletion of the pksAC prevented production of ascochitine and its derivative ascochital in A. fabae. The putative ascochitine biosynthesis gene cluster comprises 11 genes that have undergone rearrangement and gain-and-loss events relative to the citrinin biosynthesis gene cluster in Monascus ruber. Interestingly, we also identified pksAC homologs in two recently diverged species, A. lentis and A. lentis var. lathyri, that are sister taxa closely related to ascochitine producers such as A. fabae and A. viciae-villosae. However, nonsense mutations have been independently introduced in coding sequences of the pksAC homologs of A. lentis and A. lentis var. lathyri that resulted in loss of ascochitine production. Despite its reported phytotoxicity, ascochitine was not a pathogenicity factor in A. fabae infection and colonization of faba bean (Vicia faba L.). Ascochitine was mainly produced from mature hyphae at the site of pycnidial formation, suggesting a possible protective role of the compound against other microbial competitors in nature. This report highlights the evolution of gene clusters harnessing the structural diversity of polyketides and a mechanism with the potential to alter secondary metabolite profiles via single nucleotide polymorphisms in closely related fungal species. IMPORTANCE Fungi produce a diverse array of secondary metabolites, many of which are of pharmacological importance whereas many others are noted for mycotoxins, such as aflatoxin and citrinin, that can threaten human and animal health. The polyketide-derived compound ascochitine, which is structurally similar to citrinin mycotoxin, has been considered to be important for pathogenicity of legume-associated Ascochyta species. Here, we identified the ascochitine polyketide synthase (PKS) gene in Ascochyta fabae and its neighboring genes that may be involved in ascochitine biosynthesis. Interestingly, the ascochitine PKS genes in other legume-associated Ascochyta species have been mutated, encoding truncated PKSs. This indicated that point mutations may have contributed to genetic diversity for secondary metabolite production in these fungi. We also demonstrated that ascochitine is not a pathogenicity factor in A. fabae. The antifungal activities and production of ascochitine during sporulation suggested that it may play a role in competition with other saprobic fungi in nature.

scholarly journals Cloning and Heterologous Expression of the Thioviridamide Biosynthesis Gene Cluster from Streptomyces olivoviridis

2013 ◽  
Vol 79 (22) ◽  
pp. 7110-7113 ◽  
Author(s):  
Masumi Izawa ◽  
Takashi Kawasaki ◽  
Yoichi Hayakawa
Keyword(s):  
Gene Cluster ◽  
Draft Genome ◽  
Peptide Antibiotic ◽  
Heterologous Production ◽  
Gene Encoding ◽  
Biosynthesis Gene Cluster ◽  
Content Type ◽  
Precursor Peptide ◽  
Biosynthesis Gene ◽  
Draft Genome Sequencing

ABSTRACTThioviridamide is a unique peptide antibiotic containing five thioamide bonds fromStreptomyces olivoviridis. Draft genome sequencing revealed a gene (thetvaAgene) encoding the thioviridamide precursor peptide. The thioviridamide biosynthesis gene cluster was identified by heterologous production of thioviridamide inStreptomyces lividans.

scholarly journals Identification and Heterologous Expression of the Chaxamycin Biosynthesis Gene Cluster from Streptomyces leeuwenhoekii

2015 ◽  
Vol 81 (17) ◽  
pp. 5820-5831 ◽  
Author(s):  
Jean Franco Castro ◽  
Valeria Razmilic ◽  
Juan Pablo Gomez-Escribano ◽  
Barbara Andrews ◽  
Juan A. Asenjo ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Mutational Analysis ◽  
Streptomyces Coelicolor ◽  
Atacama Desert ◽  
Biosynthesis Gene Cluster ◽  
Content Type ◽  
Biosynthesis Gene ◽  
Viable Approach

ABSTRACTStreptomyces leeuwenhoekii, isolated from the hyperarid Atacama Desert, produces the new ansamycin-like compounds chaxamycins A to D, which possess potent antibacterial activity and moderate antiproliferative activity. We report the development of genetic tools to manipulateS. leeuwenhoekiiand the identification and partial characterization of the 80.2-kb chaxamycin biosynthesis gene cluster, which was achieved by both mutational analysis in the natural producer and heterologous expression inStreptomyces coelicolorA3(2) strain M1152. Restoration of chaxamycin production in a nonproducing ΔcxmKmutant (cxmKencodes 3-amino-5-hydroxybenzoic acid [AHBA] synthase) was achieved by supplementing the growth medium with AHBA, suggesting that mutasynthesis may be a viable approach for the generation of novel chaxamycin derivatives.

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