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Applied and Environmental Microbiology, February 2004, p. 686-692, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.686-692.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Genes Coding for Hepatotoxic Heptapeptides (Microcystins) in the Cyanobacterium Anabaena Strain 90

Leo Rouhiainen,1 Tanja Vakkilainen,1 Berit Lumbye Siemer,1,{dagger} William Buikema,2 Robert Haselkorn,2 and Kaarina Sivonen1*

Department of Applied Chemistry and Microbiology, Viikki Biocenter, University of Helsinki, Helsinki, Finland,1 Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois 606372

Received 17 July 2003/ Accepted 22 October 2003

The cluster of microcystin synthetase genes from Anabaena strain 90 was sequenced and characterized. The total size of the region is 55.4 kb, and the genes are organized in three putative operons. The first operon (mcyA-mcyB-mcyC) is transcribed in the opposite direction from the second operon (mcyG-mcyD-mcyJ-mcyE-mcyF-mcyI) and the third operon (mcyH). The genes mcyA, mcyB, and mcyC encode nonribosomal peptide synthetases (NRPS), while mcyD codes for a polyketide synthase (PKS), and mcyG and mcyE are mixed NRPS-PKS genes. The genes mcyJ, mcyF, and mcyI are similar to genes coding for a methyltransferase, an aspartate racemase, and a D-3-phosphoglycerate dehydrogenase, respectively. The region in the first module of mcyB coding for the adenylation domain was found to be 96% identical with the corresponding part of mcyC, suggesting a recent duplication of this fragment and a replacement in mcyB. In Anabaena strain 90, the order of the domains encoded by the genes in the two sets (from mcyG to mcyI and from mcyA to mcyC) is colinear with the hypothetical order of the enzymatic reactions for microcystin biosynthesis. The order of the microcystin synthetase genes in Anabaena strain 90 differs from the arrangement found in two other cyanobacterial species, Microcystis aeruginosa and Planktothrix agardhii. The average sequence match between the microcystin synthetase genes of Anabaena strain 90 and the corresponding genes of the other species is 74%. The identity of the individual proteins varies from 67 to 81%. The genes of microcystin biosynthesis from three major producers of this toxin are now known. This makes it possible to design probes and primers to identify the toxin producers in the environment.

* Corresponding author. Mailing address: University of Helsinki, Department of Applied Chemistry and Microbiology, P.O. Box 56, Viikki Biocenter, 00014 Helsinki University, Finland. Phone: 358-9-19159270. Fax: 358-9-19159322. E-mail: kaarina.sivonen{at}helsinki.fi.

{dagger} Present address: Danish Veterinary Laboratory, DK-1790 Copenhagen V, Denmark.

Applied and Environmental Microbiology, February 2004, p. 686-692, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.686-692.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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