Synthetic biology routes to new and extinct natural products

Abstract

Recent developments in genome sequencing and genetic engineering have revolutionised elucidation of biosynthetic pathways in bacteria and fungi and allowed production of new natural products and engineered strains with optimised production of new and/or preferred metabolites. The clinically important antibiotic mupirocin is a mixture of closely related pseudomonic acids produced by Pseudomonas fluorescens via a trans-AT modular PKS. Extensive gene knock-out experiments have led to the isolation of a plethora of new metabolites: both biosynthetic intermediates and shunt products. Parallel experiments, along with swapping of biosynthetic genes, with a Pseudoalteromonas sp. which produces the closely related thiomarinols give similar results and many new products. A genetically engineered strain of P. fluorescens produces high titres of a single pseudomonic acid with improved stability and antibiotic properties. Tenellin and bassianin are insecticidal fungal metabolites produced by Beauvaria species via multi-domain PKS-NRPSs. Heterologous expression in Aspergillus oryzae of hybrid systems produced by domain swapping between the two biosynthetic gene clusters produce many new metabolites in high yields and reveal the key elements in control of polyketide chain length and methylation, showing the potential for combinatorial biosynthesis of these and related metabolites. Cryptosporiopsis sp. 8999 produces three related dimeric xanthones. Gene knock-outs allow elucidation of the full biosynthetic pathway, isolation of the monomeric precursor and engineering of a strain producing only the major component of the wild-type mixture.