Heterologous biosynthesis of nodulisporic acid

Abstract

Nodulisporic acids (NAs) comprise a group of indole diterpenes known for their potent insecticidal activities; however, biosynthesis of NAs by its natural producer, Hypoxylon pulicicidum (Nodulisporium sp.) is exceptionally difficult to achieve. The identification of genes responsible for NA production could enable biosynthetic pathway optimization to provide access to NAs for commercial applications. Obtaining useful quantities of NAs using published fermentations methods is challenging, making gene knockout studies an undesirable method to confirm gene function. Alternatively, heterologous gene expression of H. pulicicidum genes in a more robust host species like Penicillium paxilli provides a way to rapidly identify the function of genes that play a role in NA biosynthesis. In this work, we identified the function of four secondary-metabolic genes necessary for the biosynthesis of nodulisporic acid F (NAF) and reconstituted these genes in the genome of P. paxilli to enable heterologous production of NAF in this fungus.

Description

technical field [0001] The present invention relates generally to novel polypeptides that catalyze at least one biochemical reaction that produces Nodulisporic acid (NA), polynucleotides encoding these polypeptides, methods of making these polypeptides and polynucleotides, and Methods of using these polypeptides and polynucleotides to produce at least one NA by heterologous expression in a permissive host. Background technique [0002] Filamentous fungi produce a wide variety of interesting and useful chemical compounds. Members of one such class of compounds, the indolediterpenes (IDTs), are known for their wide chemical diversity and concomitant biological activities including anti-MRSA 1 , anticancer 2，3 , anti-H1N1 4 , insecticide 5 and tremor-inducing 6 activity is of particular interest. NA( figure 1 ) is a group of quasi-paspalum-like IDTs with significant biological activity produced by Hypoxylon pulicicidum previously classified as Nodulisporium sp. 7 . Pol...

AI Technical Summary

Problems solved by technology

Due to the difficulty of NAA 10 biosynthesis in H. pulicicidum, it is challenging to obtain useful quantities of NAA 10 using published fermentation methods, and the production of NAA 10 in commercial quantities is largely impossible

Accordingly, attempts have been made to chemically synthesize NAA10, leading to the discovery of the synthetic polyarthrosporic acid F(NAF)5a 10 and polyarthic acid D 7a 11 mechanism, but the total synthesis of NAA 10 has not yet been achieved 12

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