Unravelling triterpene biosynthesis through functional characterization of an oxidosqualene cyclase (OSC) from Cleome arabica L

Abstract

Cleome arabica is a medicinal plant contains diverse bioactive compounds and terpenoids are the major components. However, the isolation and purification of the active triterpenes from this plant involve long and complicated procedures. The present work investigates the triterpenes profiles of different tissues, besides that, describes the isolation, heterologous expression and functional characterization of C. arabica gene coding for triterpenes synthases. The phytochemical investigation through GC-MS revealed significant accumulation of pentacyclic triterpenes in leaves and siliques at mature stage compared to the stems and roots of C. arabica. Among the pentacyclic triterpenes, the lupeol reached the highest level of 320 μg/g DW in leaves at maturity stage compared to the other tissues. The biosynthesis of a pentacyclic triterpene was investigated through isolation and cloning of a full-length oxidosqualene cyclase cDNA (CaOSC) from mature leaves of C. arabica. The bioinformatic analyses revealed that CaOSC was highly homologous with the characterized lupeol synthases and shared 79.3% identity to camelliol C synthase from A. thaliana. Heterologous expression of CaOSC gene in Saccharomyces cerevisiae synthesized lupeol as a single product. The lupeol biosynthesis was exponentially increased after induction through the fermentation process reaching the maximum of 2.33 μg/ml for 240 h. Furthermore, organ-specific expression of lupeol gene was exactly matched the accumulation pattern in different tissues of C. arabica during phenological cycle. Thus, the identified CaOSC will be useful in enhancing triterpene yield for industrial purposes.

Keywords: Cleome arabica; Heterologous expression; Lupeol; Oxidosqualene cyclase; Pentacyclic triterpene.