Targeted Genome Mining Reveals the Biosynthetic Gene Clusters of Natural Product CYP51 Inhibitors

Abstract

Lanosterol 14α-demethylase (CYP51) is an important target in the development of antifungal drugs. The fungalderived restricticin 1 and related molecules are the only examples of natural products that inhibit CYP51. Here, using colocalizations of genes encoding self-resistant CYP51 as the query, we identified and validated the biosynthetic gene cluster (BGC) of 1. Additional genome mining of related BGCs with CYP51 led to production of the related lanomycin 2. The pathways for both 1 and 2 were identified from fungi not known to produce these compounds, highlighting the promise of the self-resistance enzyme (SRE) guided approach to bioactive natural product discovery.

Biosynthesis of the Fusarium Mycotoxin (−)-Sambutoxin

“…Compound 7 was structurally confirmed to be the 2-pyridone ketone (Table S6 and Figures S6A, S12–S16), consistent with the functions of P450 RE enzymes in other 2-pyridone alkaloid pathways. − , In addition to 7 , a cometabolite 7 ′ with +16 mu was also isolated and characterized (Figure B, Table S7, and Figures S17–S21). Compound 7 ′ was found to contain a secondary alcohol in the polyketide chain of 7 , which may be attributed to the activity of endogenous alkyl hydroxylases in A. nidulans observed in other reconstitution studies. , It is worth noting here that 7 ′ persists throughout the remaining reconstitution work (Figure S3) and is not modified by the downstream smb enzymes.…”

Deciphering chemical logic of fungal natural product biosynthesis through heterologous expression and genome mining

“…In this example, even though restricticin was not isolated as a pure compound from the heterologous host, formation of acetylated and modied versions of the compound conrmed the role of the BGC, which led to identication of additional CYP51 inhibitors such as lanomycin using genome mining approaches. 166 This journal is © The Royal Society of Chemistry 2023 167 However, it is challenging to control stereoselectivity, regioselectivity and periselectivity of pericyclic reactions without catalysts, especially when competing transition states are accessible. In natural product biosynthesis, these types of selectivity are strictly controlled by pericyclases, a family of enzymes that catalyze pericyclic reactions.…”

Biosynthesis of fungal polyketides by collaborating and trans-acting enzymes

“…70,184 However, the main reason for the complexity in this pathway is that CazF is capable of synthesizing two different triketides 108 and 109 ; 108 is transferred to CazM, and 109 is transferred by CazE to the modified product of CazF/CazM cazisochromene 110 (Scheme 21B). 169 This is a revealing example of collaborating enzymes competing for substrates and altering the product profile of a megasynthase.…”