Engineering Critical Enzymes and Pathways for Improved Triterpenoid Biosynthesis in Yeast

Abstract

Triterpenoids represent a diverse group of phytochemicals, widely distributed in the plant kingdom with many biological activities. Recently, the heterologous production of triterpenoids in Saccharomyces cerevisiae has been successfully implemented by introducing various triterpenoids biosynthetic pathways. By engineering related enzymes as well as yeast metabolism, the yield of various triterpenoids is significantly improved from milligram-scale per liter to gram-scale level per liter. This achievement demonstrates that engineering of critical enzymes is considered as a potential strategy to overcome the main hurdles of translation of these potent natural products into industry. Here, we review strategies for designing enzymes to improve the yield of triterpenoids in S. cerevisiae, which is mainly separated into three aspects: 1. elevating the supply of the precursor-2,3-oxidosqualene, 2. optimizing triterpenoid-involved reactions, 3. lowering the competition of the native sterol pathway. And then we provide challenges and prospects on further enhancing the triterpenoid production in S. cerevisiae.

High-Level Production of Sesquiterpene Patchoulol in Saccharomyces cerevisiae

“…They show numerous vital functions in all organisms and can be used in various applications, ranging from foods to cosmetics, and are an important source of valuable medicinal compounds. , Usually, in nature, terpenes or terpenoids are produced in small quantities. Chemical synthesis of these compounds is very difficult because of their complex structures, including their chirality. , Alternative methods have been explored for terpene/terpenoid production due to the importance and great demand. ,, In the past 10 years, advances in metabolic engineering and synthetic biology have accelerated the construction of microbial cell factories to obtain a sustainable supply of limited natural products, such as artemisinic acid, taxadiene, ginsenosides, miltiradiene, alkaloids, and cannabinoids. − ,− …”

“…2,3 Alternative methods have been explored for terpene/terpenoid production due to the importance and great demand. 1,4,5 In the past 10 years, advances in metabolic engineering and synthetic biology have accelerated the construction of microbial cell factories to obtain a sustainable supply of limited natural products, such as artemisinic acid, taxadiene, ginsenosides, miltiradiene, alkaloids, and cannabinoids. 1−3,6−9 Sesquiterpene patchoulol, which is found in the leaves of patchouli plants, is an important compound due to its potent pharmacological properties and characteristic aromas and flavours and is extensively used in the perfume and fragrance industry.…”

Pentacyclic Triterpenoid Saponins, Poterinasides A–J, from Silverweed Cinquefoil Roots Promising Hepatoprotective and Anti-inflammatory Agents

“…The plausible biosynthetic pathway of compounds 1 and 2 is postulated as shown in Scheme . Triterpenoid aglycone backbones are derived from the linear 30-carbon precursor squalene, which is oxidized to hydroxyl oxidosqualene by a series of enzymes (squalene epoxidase, SE; cytochrome P450, CYP; cytochrome P450 reductase, CPR) . Hydroxyl oxidosqualene is subsequently transformed into dammarenyl cation under acidic conditions by dammarenediol II synthase (DDS) and dehydration reactions .…”

Metabolic Engineering of Saccharomyces cerevisiae for High-Level Friedelin via Genetic Manipulation

“…However, the supply of friedelin from plant sources is insufficient, while chemical methods are often complex, include extreme reaction conditions, and produce toxic chemicals; thus, it is urgent to develop an environmentally friendly and cost-effective method of securing the friedelin supply (Moses et al, 2013;Guo et al, 2020). Synthetic biology and metabolic engineering have provided promising and green approaches to reconstruct microorganisms to yield these natural high-value products.…”