A bacterial hormone (the SCB1) directly controls the expression of a pathway-specific regulatory gene in the cryptic type I polyketide biosynthetic gene cluster of Streptomyces coelicolor

Streptomyces antibiotic regulatory proteins (SARPs) have been shown to activate transcription by binding to a tandemly arrayed set of heptameric direct repeats located around the −35 region of their cognate promoters. Experimental evidence is presented here showing that vlmI is a regulatory gene in the valanimycin biosynthetic gene cluster of Streptomyces viridifaciens and encodes a protein belonging to the SARP family. The organization of the valanimycin biosynthetic gene cluster suggests that the valanimycin biosynthetic genes are located on three potential transcripts, vlmHORBCD, vlmJKL and vlmA. Disruption of vlmI abolished valanimycin biosynthesis. Western blot analyses showed that VlmR and VlmA are absent from the vlmI mutant and that the production of VlmK is severely diminished. These results demonstrate that the expression of these genes from the three potential transcripts is under the positive control of VlmI. The vlmA–vlmH and vlmI–vlmJ intergenic regions both exhibit a pattern of heptameric direct repeats. Gel shift assays with VlmI overproduced in Escherichia coli as a C-terminal FLAG-tagged protein clearly demonstrated that VlmI binds to DNA fragments from both regions that contain these heptameric repeats. When a high-copy-number vlmI expression plasmid was introduced into Streptomyces coelicolor M512, which contains mutations in the undecylprodigiosin and actinorhodin activators redD and actII-orf4, undecylprodigiosin production was restored, showing that vlmI can complement a redD mutation. Introduction of the same vlmI expression plasmid into an S. viridifaciens vlmI mutant restored valanimycin production to wild-type levels.

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Identification of the Anti-Infective Aborycin Biosynthetic Gene Cluster from Deep-Sea-Derived Streptomyces sp. SCSIO ZS0098 Enables Production in a Heterologous Host

Marine Drugs ◽

10.3390/md17020127 ◽

2019 ◽

Vol 17 (2) ◽

pp. 127 ◽

Cited By ~ 6

Author(s):

Mingwei Shao ◽

Juying Ma ◽

Qinglian Li ◽

Jianhua Ju

Keyword(s):

Antibacterial Activity ◽

Gene Cluster ◽

Deep Sea ◽

Streptomyces Coelicolor ◽

Biosynthetic Gene Cluster ◽

Type I ◽

Biosynthetic Gene ◽

Heterologous Host ◽

Streptomyces Sp ◽

Enterococcus Gallinarum

Aborycin is a ribosomally synthesized member of the type I lasso peptide natural products. In the present study, aborycin was isolated and identified from the deep-sea-derived microbe Streptomyces sp. SCSIO ZS0098. The aborycin biosynthetic gene cluster (abo) was identified on the basis of genome sequence analyses and then heterologously expressed in Streptomyces coelicolor M1152 to effectively produce aborycin. Aborycin generated in this fashion exhibited moderate antibacterial activity against 13 Staphylococcus aureus strains from various sources with minimum inhibitory concentrations MICs = 8.0~128 µg/mL, against Enterococcus faecalis ATCC 29212 with an MIC = 8.0 µg/mL, and against Bacillus thuringiensis with MIC = 2.0 µg/mL. Additionally, aborycin displayed potent antibacterial activity (MIC = 0.5 µg/mL) against the poultry pathogen Enterococcus gallinarum 5F52C. The reported abo cluster clearly has the potential to provide a means of expanding the repertoire of anti-infective type I lasso peptides.

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Identification and Heterologous Expression of the Kendomycin B Biosynthetic Gene Cluster from Verrucosispora sp. SCSIO 07399

Marine Drugs ◽

10.3390/md19120673 ◽

2021 ◽

Vol 19 (12) ◽

pp. 673

Author(s):

Jiang Chen ◽

Shanwen Zhang ◽

Yingying Chen ◽

Xinpeng Tian ◽

Yucheng Gu ◽

...

Keyword(s):

Heterologous Expression ◽

Gene Cluster ◽

Polyketide Synthase ◽

Regulatory Gene ◽

Biosynthetic Gene Cluster ◽

Chain Elongation ◽

Type I ◽

Biosynthetic Gene ◽

Type Iii Polyketide Synthase ◽

Type Iii

Verrucosispora sp. SCSIO 07399, a rare marine-derived actinomycete, produces a set of ansamycin-like polyketides kendomycin B–D (1–3) which possess potent antibacterial activities and moderate tumor cytotoxicity. Structurally, kendomycin B–D contain a unique aliphatic macrocyclic ansa scaffold in which the highly substituted pyran ring is connected to the quinone moiety. In this work, a type I/type III polyketide synthase (PKS) hybrid biosynthetic gene cluster coding for assembly of kendomycin B (kmy), and covering 33 open reading frames, was identified from Verrucosispora sp. SCSIO 07399. The kmy cluster was found to be essential for kendomycin B biosynthesis as verified by gene disruption and heterologous expression. Correspondingly, a biosynthetic pathway was proposed based on bioinformatics, cluster alignments, and previous research. Additionally, the role of type III PKS for generating the precursor unit 3,5-dihydroxybenzoic acid (3,5-DHBA) was demonstrated by chemical complementation, and type I PKS executed the polyketide chain elongation. The kmy cluster was found to contain a positive regulatory gene kmy4 whose regulatory effect was identified using real-time quantitative PCR (RT-qPCR). These advances shed important new insights into kendomycin B biosynthesis and help to set the foundation for further research aimed at understanding and exploiting the carbacylic ansa scaffold.

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Investigation of the effects of actinorhodin biosynthetic gene cluster expression and a rpoB point mutation on the metabolome of Streptomyces coelicolor M1146

Journal of Bioscience and Bioengineering ◽

10.1016/j.jbiosc.2021.01.002 ◽

2021 ◽

Author(s):

Katsuaki Nitta ◽

Rainer Breitling ◽

Eriko Takano ◽

Sastia P. Putri ◽

Eiichiro Fukusaki

Keyword(s):

Gene Cluster ◽

Point Mutation ◽

Streptomyces Coelicolor ◽

Biosynthetic Gene Cluster ◽

Biosynthetic Gene

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New Insights into the Genetic Organization of the FK228 Biosynthetic Gene Cluster inChromobacterium violaceumNo. 968

Applied and Environmental Microbiology ◽

10.1128/aem.01512-10 ◽

2010 ◽

Vol 77 (4) ◽

pp. 1508-1511 ◽

Cited By ~ 13

Author(s):

Vishwakanth Y. Potharla ◽

Shane R. Wesener ◽

Yi-Qiang Cheng

Keyword(s):

Natural Product ◽

Gene Cluster ◽

Gene Deletion ◽

Regulatory Gene ◽

Biosynthetic Gene Cluster ◽

Transcriptional Analysis ◽

Biosynthetic Gene ◽

Genetic Organization

ABSTRACTThe biosynthetic gene cluster of FK228, an FDA-approved anticancer natural product, was identified and sequenced previously. The genetic organization of this gene cluster has now been delineated through systematic gene deletion and transcriptional analysis. As a result, the gene cluster is redefined to contain 12 genes:depAthroughdepJ,depM, and a newly identified pathway regulatory gene,depR.

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Cloning and characterization of the histidine biosynthetic gene cluster of Streptomyces coelicolor A3(2)

Gene ◽

10.1016/0378-1119(90)90436-u ◽

1990 ◽

Vol 90 (1) ◽

pp. 31-41 ◽

Cited By ~ 22

Author(s):

Danila Limauro ◽

Alessandra Avitabile ◽

Carmela Cappellano ◽

Anna Maria Puglia ◽

Carmelo B. Bruni

Keyword(s):

Gene Cluster ◽

Streptomyces Coelicolor ◽

Biosynthetic Gene Cluster ◽

Biosynthetic Gene

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Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host

Marine Drugs ◽

10.3390/md17100560 ◽

2019 ◽

Vol 17 (10) ◽

pp. 560 ◽

Cited By ~ 5

Author(s):

Zhijie Yang ◽

Xin Wei ◽

Jianqiao He ◽

Changli Sun ◽

Jianhua Ju ◽

...

Keyword(s):

Gene Cluster ◽

Streptomyces Coelicolor ◽

Regulatory Gene ◽

Regulatory Protein ◽

Biosynthetic Gene Cluster ◽

Negative Regulator ◽

Over Expression ◽

Lead Compounds ◽

Mycobacteria Tuberculosis ◽

Abc Transporter Genes

Atratumycin is a cyclodepsipeptide with activity against Mycobacteria tuberculosis isolated from deep-sea derived Streptomyces atratus SCSIO ZH16NS-80S. Analysis of the atratumycin biosynthetic gene cluster (atr) revealed that its biosynthesis is regulated by multiple factors, including two LuxR regulatory genes (atr1 and atr2), two ABC transporter genes (atr29 and atr30) and one Streptomyces antibiotic regulatory gene (atr32). In this work, three regulatory and two transporter genes were unambiguously determined to provide positive, negative and self-protective roles during biosynthesis of atratumycin through bioinformatic analyses, gene inactivations and trans-complementation studies. Notably, an unusual Streptomyces antibiotic regulatory protein Atr32 was characterized as a negative regulator; the function of Atr32 is distinct from previous studies. Five over-expression mutant strains were constructed by rational application of the regulatory and transporter genes; the resulting strains produced significantly improved titers of atratumycin that were ca. 1.7–2.3 fold greater than wild-type (WT) producer. Furthermore, the atratumycin gene cluster was successfully expressed in Streptomyces coelicolor M1154, thus paving the way for the transfer and recombination of large DNA fragments. Overall, this finding sets the stage for understanding the unique biosynthesis of pharmaceutically important atratumycin and lays the foundation for generating anti-tuberculosis lead compounds possessing novel structures.

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