Cytochromes P450 for Terpene Functionalisation and Metabolic Engineering

Abstract

Plants have evolved the capacity to produce a striking array of specialised metabolites. Terpenoids are the oldest and most diverse class of such compounds and have attracted interest for industrial and pharmaceutical applications. The development of biotechnological alternatives for their production is the focus of intense research. Photosynthetic systems provide new strategies for autotrophic metabolic engineering. Focusing on cytochromes P450, involved in the functionalisation of the core terpene molecules, this review highlights the latest approaches in this field and looks towards recent discoveries that have the potential to shape the future of terpenoid bioengineering.

Carnosic acid biosynthesis elucidated by a synthetic biology platform

“…In the case study presented here, metabolomic and transcriptomic analysis was combined in the same plants (37). Following next-generation sequencing, genes were prioritized for analysis on the basis of their transcription levels and by increasing the representation of the CYP71 and CYP76 subfamilies, members of which have previously been found to be involved in terpene oxidation (29). Rational short listing of candidate genes can help minimize time and reduce cost.…”

“…20). Subsequent decoration of the various skeletons by modifying enzymes, such as cytochrome P450s (CYPs) (27), gives rise to a plethora of secondary metabolites (28,29). In Rosmarinus officinalis and Salvia fruticosa, carnosic acid and carnosol are synthesized from (+)-copalyl diphosphate (CPP; 13) through the intermediates miltiradiene (14), abietatriene (15), and ferruginol (16) (Fig.…”

Photosynthetic fuel for heterologous enzymes: the role of electron carrier proteins

“…It has been demonstrated for only a handful of enzymes (Table 1), all of which are cytochrome P450s. Cytochrome P450s metabolize drugs and toxins in humans (Nebert et al 2013) and feature prominently in the biosynthesis of plant specialized metabolites of commercial interest such as terpenoids, alkaloids, and polyphenols (Renault et al 2014;Pateraki et al 2015). Consequently, they attract considerable attention for use in biotechnology and are frequently expressed in heterologous organisms and engineered for improved activity and recognition of new substrates (Fasan 2012;Lassen et al 2014b).…”

Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii

“…All terpenes are derived from the five carbon containing building blocks isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), which are sequentially condensed by the diphosphate synthases (PPs, also known as prenyl transferases): geranyl diphosphate synthase to geranyl diphosphate (GPP, C10), farnesyl diphosphate synthase to farnesyl diphosphate (FPP, C15), and geranylgeranyl diphosphate synthase to geranylgeranyl diphosphate (GGPP, C20). These carbon precursors are rapidly converted into the vast diversity of known terpenoids by a class of enzymes known as terpene synthases, and the resulting hydrocarbon skeletons can be further modified, for example, by cyclization or addition of functional groups by cytochrome P450 enzymes (Pateraki et al, 2015;Weitzel and Simonsen, 2013).…”