Directing the Heterologous Production of Specific Cyanobacterial Toxin Variants,ACS Chemical Biology

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Directing the Heterologous Production of Specific Cyanobacterial Toxin Variants
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2017-06-01 00:00:00 , DOI: 10.1021/acschembio.7b00181
Tianzhe Liu ₁ , Rabia Mazmouz _{1,

2} , Sarah E. Ongley _{1,

2} , Rocky Chau ₁ , Russell Pickford ₃ , Jason N. Woodhouse _{1,

4} , Brett A. Neilan _{1,

2}

Affiliation

Microcystins are globally the most commonly occurring freshwater cyanotoxins, causing acute poisoning and chronically inducing hepatocellular carcinoma. However, the detection and toxicological study of microcystins is hampered by the limited availability and high cost of pure toxin standards. Biosynthesis of microcystin variants in a fast-growing heterologous host offers a promising method of achieving reliable and economically viable alternative to isolating toxin from slow-growing cyanobacterial cultures. Here, we report the heterologous expression of recombinant microcystin synthetases in Escherichia coli to produce [d-Asp³]microcystin-LR and microcystin-LR. We assembled a 55 kb hybrid polyketide synthase/nonribosomal peptide synthetase gene cluster from Microcystis aeruginosa PCC 7806 using Red/ET recombineering and replaced the native promoters with an inducible Ptet_O promoter to yield microcystin titers superior to M. aeruginosa. The expression platform described herein can be tailored to heterologously produce a wide variety of microcystin variants, and potentially other cyanobacterial natural products of commercial relevance.

中文翻译：

指导异源蓝藻毒素变种的异源生产

微囊藻毒素是全球最常见的淡水蓝藻毒素，可引起急性中毒并长期诱发肝细胞癌。然而，微囊藻毒素的检测和毒理学研究由于纯毒素标准品的有限可用性和高成本而受到阻碍。快速增长的异源宿主中微囊藻毒素变体的生物合成提供了一种有前途的方法，可实现从慢速生长的蓝细菌培养物中分离毒素的可靠且经济可行的选择。在这里，我们报告在大肠杆菌中产生[ d -Asp ³ ] microcystin-LR和microcystin-LR的重组微囊藻毒素合成酶的异源表达。我们组装了一个55 kb的杂合聚酮化合物合酶/非核糖体肽合成酶基因簇铜绿微囊藻PCC 7806使用Red / ET重组，并用可诱导的P tet _O启动子代替了天然启动子，产生的微囊藻毒素滴度优于铜绿假单胞菌。可将本文所述的表达平台定制为异源产生多种微囊藻毒素变体，以及可能具有商业意义的其他蓝细菌天然产物。

更新日期：2017-06-28

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