- Title
- Mutagenesis of the microcystin tailoring and transport proteins in a heterologous cyanotoxin expression system
- Creator
- Liu, Tianzhe; Mazmouz, Rabia; Pearson, Leanne A.; Neilan, Brett A.
- Relation
- ARC.LP140100642 http://purl.org/au-research/grants/arc/LP140100642
- Relation
- ACS Synthetic Biology Vol. 8, Issue 5, p. 1187-1194
- Publisher Link
- http://dx.doi.org/10.1021/acssynbio.9b00068
- Publisher
- American Chemical Society (ACS)
- Resource Type
- journal article
- Date
- 2019
- Description
- The microcystins are a large group of cyclic peptide hepatotoxins produced by several genera of freshwater cyanobacteria. The genes responsible for microcystin biosynthesis are encoded within a large (~55 kbp) gene cluster, mcyA-J. The recent establishment of a cyanotoxin heterologous expression system in Escherichia coli has provided the means to study microcystin biosynthesis in a genetically tractable, rapidly growing host. Using this system, we demonstrate that deletion of the ABC-transporter, mcyH, and dehydrogenase, mcyI, abolishes microcystin production, while deletion of the O-methyltransferase, mcyJ, results in the production of the demethylated (DM) toxin [D-Asp3 , DMAdda 5]microcystin-LR. Both methylated and DM toxin variants were heterologously produced at high titers and efficiently exported into the extracellular medium, enabling easy purification. The results show that the mcy gene cluster can be engineered in E. coli to study the function of its individual components and direct the synthesis of particular microcystin variants. This technology could potentially be applied to other natural products of ecological and biomedical significance.
- Subject
- cyanobacteria; hepatotoxin; microcystin; nonribosomal peptide; polyketide; mutagenesis; biosynthetic gene cluster
- Identifier
- http://hdl.handle.net/1959.13/1470599
- Identifier
- uon:48515
- Identifier
- ISSN:2161-5063
- Language
- eng
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