Biosynthesis of Novel Statins by Combining Heterologous Genes from Xylaria and Aspergillus

Abstract

For many secondary metabolites, heterologous synthesis is the definitive step to determine their required biosynthetic genes. Using a multivector expression system in Saccharomyces cerevisiae, we reconstituted not only two natural statins from two fungal species, i.e., lovastatin from Aspergillus terreus and FR901512 from Xylaria grammica, but also new statin structures by mixing their genes. Combinatorial gene exchange experiments revealed the functional promiscuity of two polyketide synthases in A. terreus, lovB, and lovF; they could synthesize FR901512 with Xylaria genes. Key structure determinants of statins are essential accessory genes that are irreplaceable across species.

Recent progress in biodiversity research on the Xylariales and their secondary metabolism

“…Within the same work reporting on a multitude of new terpenoids from a species potentially representing H. fendleri (see Terpenoids section), several terphenyls were reported [89]. Apart from rickenyls C-E known from H. rickii [98], four new terphenyls named fendleryls A (69) to D were isolated [89]. Fendleryls A−D exhibited no antimicrobial activity, but weak cytotoxicity against NCI-H187 and Vero cells with IC 50 values of 48 and 49 µM, respectively.…”

“…Another approach to attain novel natural products is heterologous gene expression. Recently, this concept has been utilized by mixing genes responsible for production of the potent statins (of which lovastatin is the most famous representative) from two producing organisms, Aspergillus terreus and X. grammica, in Saccharomyces cerevisiae [69]. This ultimately resulted in MS/MS predictions of two novel structures, O-acetylmonacolin J and "methylbutyryl DA_FR901512".…”

Statins—From Fungi to Pharmacy

“…Moreover, the binding is also determined by the compound structure; e.g., the base structure of synthetic statins (i.e., cerivastatin, fluvastatin, atorvastatin, and rosuvastatin) containing fluorinated phenol groups provides additional sites for enzyme binding. Nevertheless, the differences in the structure are thought to impact statins’ bioactivity, determining their medical values, while the mechanistic relationship between medicinal properties and core structure has not been elucidated [ 61 ]. Evaluation of the crystal structures of the statin-enzyme complex showed that statins bind with HMGR with a large number of van der Waals forces.…”

Expanding the Substrate Scope of Acyltransferase LovD9 for the Biosynthesis of Statin Analogues

“…[1] Thus, a previous research focused on heterologous genes combination for novel statin biosynthesis. [2] Another work explored the substrate scope of wild-type acyltransferase LovD, which naturally acylates Monacolin J acid (MJA) to yield lovastatin aided by an acyl carrier protein (LovF-ACP). [3] Synthetic surrogates, such as acyl-CoA, N-acetyl cysteamine (SNAC) and methyl thioglycolate (SMTG) thioesters were successfully used as acyl donors, substituting the S-acyl-LovF native ACP.…”

“…These compounds, which belong to the family of statins, entail a big pharmaceutical value as cholesterol‐lowering drugs since they inhibit hydroxy methyl glutaryl coenzyme A (HMG‐CoA) reductase [1] . Thus, a previous research focused on heterologous genes combination for novel statin biosynthesis [2] . Another work explored the substrate scope of wild‐type acyltransferase LovD, which naturally acylates Monacolin J acid (MJA) to yield lovastatin aided by an acyl carrier protein (LovF‐ACP) [3] .…”