Microcyclamide Biosynthesis in Two Strains of Microcystis aeruginosa: from Structure to Genes and Vice Versa

ABSTRACT Comparative analysis of related biosynthetic gene clusters can provide new insights into the versatility of these pathways and allow the discovery of new natural products. The freshwater cyanobacterium Microcystis aeruginosa NIES298 produces the cytotoxic peptide microcyclamide. Here, we provide evidence that the cyclic hexapeptide is formed by a ribosomal pathway through the activity of a set of processing enzymes closely resembling those recently shown to be involved in patellamide biosynthesis in cyanobacterial symbionts of ascidians. Besides two subtilisin-type proteases and a heterocyclization enzyme, the gene cluster discovered in strain NIES298 encodes six further open reading frames, two of them without similarity to enzymes encoded by the patellamide gene cluster. Analyses of genomic data of a second cyanobacterial strain, M. aeruginosa PCC 7806, guided the discovery and structural elucidation of two novel peptides of the microcyclamide family. The identification of the microcyclamide biosynthetic genes provided an avenue by which to study the regulation of peptide synthesis at the transcriptional level. The precursor genes were strongly and constitutively expressed throughout the growth phase, excluding the autoinduction of these peptides, as has been observed for several peptide pheromone families in bacteria.

[1]  E. Dittmann,et al.  Biosynthesis and structure of aeruginoside 126A and 126B, cyanobacterial peptide glycosides bearing a 2-carboxy-6-hydroxyoctahydroindole moiety. , 2007, Chemistry & biology.

[2]  Lik Tong Tan,et al.  Bioactive natural products from marine cyanobacteria for drug discovery. , 2007, Phytochemistry.

[3]  H. Gross.  Strategies to unravel the function of orphan biosynthesis pathways: recent examples and future prospects , 2007, Applied Microbiology and Biotechnology.

[4]  Roger G. Linington,et al.  Venturamides A and B: antimalarial constituents of the panamanian marine Cyanobacterium Oscillatoria sp. , 2007, Journal of natural products.

[5]  Jacques Ravel,et al.  Natural combinatorial peptide libraries in cyanobacterial symbionts of marine ascidians , 2006, Nature chemical biology.

[6]  E. Breukink.  A lesson in efficient killing from two‐component lantibiotics , 2006, Molecular microbiology.

[7]  M. Welker,et al.  Cyanobacterial peptides - nature's own combinatorial biosynthesis. , 2006, FEMS microbiology reviews.

[8]  S. Sudek,et al.  Structure of Trichamide, a Cyclic Peptide from the Bloom-Forming Cyanobacterium Trichodesmium erythraeum, Predicted from the Genome Sequence , 2006, Applied and Environmental Microbiology.

[9]  P. Long,et al.  Spontaneity in the patellamide biosynthetic pathway. , 2006, Organic & biomolecular chemistry.

[10]  P. Long,et al.  Shotgun Cloning and Heterologous Expression of the Patellamide Gene Cluster as a Strategy to Achieving Sustained Metabolite Production , 2005, Chembiochem : a European journal of chemical biology.

[11]  J. Eisen,et al.  Patellamide A and C biosynthesis by a microcin-like pathway in Prochloron didemni, the cyanobacterial symbiont of Lissoclinum patella. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[12]  R. Evans,et al.  Applications of the bacteriocin, nisin , 1996, Antonie van Leeuwenhoek.

[13]  V. Nizet,et al.  Identification of a Streptolysin S-Associated Gene Cluster and Its Role in the Pathogenesis of Streptococcus iniae Disease , 2002, Infection and Immunity.

[14]  K. Harada,et al.  Simultaneous detection and determination of the absolute configuration of thiazole-containing amino acids in a peptide , 2002 .

[15]  F. Jüttner,et al.  Nostocyclamide M: a cyanobacterial cyclic peptide with allelopathic activity from Nostoc 31. , 2001, Phytochemistry.

[16]  J. Michiels,et al.  Processing and export of peptide pheromones and bacteriocins in Gram-negative bacteria. , 2001, Trends in microbiology.

[17]  E. Dittmann,et al.  Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806: an integrated peptide-polyketide synthetase system. , 2000, Chemistry & biology.

[18]  K. Ishida,et al.  Microcyclamide, a cytotoxic cyclic hexapeptide from the cyanobacterium Microcystis aeruginosa. , 2000, Journal of natural products.

[19]  R. Haselkorn,et al.  Genes encoding synthetases of cyclic depsipeptides, anabaenopeptilides, in Anabaena strain 90 , 2000, Molecular microbiology.

[20]  G. Jung,et al.  Lantibiotics and microcins: polypeptides with unusual chemical diversity. , 2000, Current opinion in chemical biology.

[21]  S. Rebuffat,et al.  The cyclic structure of microcin J25, a 21-residue peptide antibiotic from Escherichia coli. , 2001, European journal of biochemistry.

[22]  N. Kelleher,et al.  ATP/GTP hydrolysis is required for oxazole and thiazole biosynthesis in the peptide antibiotic microcin B17. , 1998, Biochemistry.

[23]  S. Carmeli,et al.  Tenuecyclamides A-D, cyclic hexapeptides from the cyanobacterium Nostoc spongiaeforme var. tenue. , 1998, Journal of natural products.

[24]  Michiel Kleerebezem,et al.  Quorum sensing by peptide pheromones and two‐component signal‐transduction systems in Gram‐positive bacteria , 1997, Molecular microbiology.

[25]  M. Asayama,et al.  Restriction barrier composed of an extracellular nuclease and restriction endonuclease in the unicellular cyanobacterium Microcystis sp. , 1996, FEMS microbiology letters.

[26]  C. Smith,et al.  Dendroamides, new cyclic hexapeptides from a blue-green alga. Multidrug-resistance reversing activity of dendroamide A. , 1996, Journal of natural products.

[27]  J. L. San Millán,et al.  Structure and organization of plasmid genes required to produce the translation inhibitor microcin C7 , 1995, Journal of bacteriology.

[28]  J. Weckesser,et al.  Cyanopeptolins, new depsipeptides from the cyanobacterium Microcystis sp. PCC 7806. , 1993, The Journal of antibiotics.

[29]  W. D. de Vos,et al.  University of Groningen Characterization of the Lactococcus lactis Nisin A Operon Genes nisP , Encoding a Subtilisin-Like Serine Protease Involved in Precursor Processing , and nisR , Encoding a Regulatory Protein Involved in Nisin Biosynthesis , 2019 .

[30]  J. Weckesser,et al.  Two Closely Related Peptide Toxins in Axenically Grown Microcystis aeruginosa PCC 7806 , 1990 .

[31]  B. Degnan,et al.  Novel cytotoxic compounds from the ascidian Lissoclinum bistratum. , 1989, Journal of medicinal chemistry.

[32]  K. Entian,et al.  Prepeptide sequence of epidermin, a ribosomally synthesized antibiotic with four sulphide-rings , 1988, Nature.

[33]  C. Franche,et al.  [88] Tests on nif probes and DNA hybridizations , 1988 .

[34]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[35]  J. Waterbury,et al.  Generic assignments, strain histories, and properties of pure cultures of cyanobacteria , 1979 .