Total and engineered biosynthesis of cinatrins: inhibitors of phospholipase A<sub>2</sub> and squalene synthase

An alkyl citrate biosynthetic gene cluster (<i>ctr</i>) was identified from 47 members of the fungal family of Hypoxylaceae by genome mining. The <i>Hypomontagnella monticulosa</i> MUCL 54604 <i>ctr</i> cluster was deployed for the total biosynthesis of the known specialised metabolites CJ-13,982 and cinatrins C₁ and C₃. The required 2- and 4- oxygenations were catalysed by an α-ketoglutarate-dependent non-heme iron dioxygenase that is homologous to known oxygenases including MfR1 and MfR2 from the squalestatin biosynthetic pathway. Combinatorial biosynthesis, and <i>in vitro</i> experiments including the oxygenases MfR1 and MfR2 from the squalestatin S1 biosynthetic gene cluster, were used for the synthesis of new natural products with bioactivity against phospholipase A₂ and squalene synthase.