Construction of yeast producing patchoulol by global metabolic engineering strategy

Abstract

Patchoulol is a sesquiterpene alcohol found in the leaves of the patchouli plant that can be extracted by steam distillation. Notably, patchoulol is an essential natural product frequently used in the chemical industry. However, patchouli produces an insignificant amount of patchoulol, not to mention steam distillation, and requires a lot of energy and time. Recombinant microorganisms that can be cultured in mild conditions and can produce patchoulol from renewable biomass resources may be a promising alternative. We previously developed the global metabolic engineering strategy (GMES), which produces a comprehensive metabolic modification in yeast, using the cocktail δ‐integration method. In this study, we aimed to produce patchoulol by modifying engineered yeast. The expression of nine genes involved in patchoulol synthesis was modulated using GMES. Regarding patchoulol production, the resultant strain, YPH499/PAT167/MVA442, showed a concentration of 42.1 mg/L, a production rate of 8.42 mg/L/d, and a yield of 2.05 mg/g‐glucose, respectably. These concentration values, production rate, and yield obtained through batch‐fermentation in this study were high level when compared to previously reported recombinant microorganism studies. GMES could be used as a potential strategy for producing secondary metabolites from plants in recombinant Saccharomyces cerevisiae.

High-Level Production of Sesquiterpene Patchoulol in Saccharomyces cerevisiae

“…Unfortunately, limited patchoulol is produced in patchouli. Moreover, the cultivation of patchouli is restricted to specific geographical locations and climate conditions, and its steam-distillation course is relatively extensive and requires much time, energy, and kerosene. , To date, several studies have reported production of patchoulol using recombinant microbial cells, including Saccharomyces cerevisiae and Corynebacterium glutamicum. , Asadollahi et al achieved patchoulol production of 11.5 mg/L by suppressing ERG9 in S. cerevisiae . Albertsen et al expressed a fused patchoulol synthase (PTS) and farnesyl pyrophosphate (FPP) synthase (FPPS) gene and suppressed ERG9 in S. cerevisiae , obtaining 40.9 mg/L patchoulol in a jar fermenter .…”

“…Henke et al produced patchoulol by overexpressing PTS and its precursor gene and suppressing the byproduct pathway in C. glutamicum , resulting in 60 mg/L of patchoulol (fed-batch fermentation) . Mitsui et al developed a global metabolic engineering strategy to obtain 42.1 mg/L patchoulol through batch fermentation . In our previous work, we constructed an engineered S. cerevisiae for production of patchoulol with a titer of 466.8 mg/L, the highest yield ever reported, in fed-batch fermentation .…”

Current trends in essential oil (EO) production

“…There are geographical locations facing climatic disturbances which generate supply problems of EOs [161]. The oil extraction from Patchouli plant could be successfully conducted using metabolic engineering technique [162].…”

Coexpression Analysis Identified PcMYB25 as a Patchoulol Synthase Gene Activator to Enhance Patchouli Alcohol Biosynthesis in Pogostemon Cablin

“…Because of patchouli alcohol's great application and commercial value, increasing the yield of patchouli alcohol by genetic engineering has become a hot research area. Mitsui et al expressed the fusion proteins of FPPS and PTS in yeast, ultimately yielding 8.42 mg/L/d (Mitsui et al 2020). However, the biosynthesis of terpenoids is a complicated process and is affected by many rate-limiting enzymes.…”