Biosynthesis of squalene-type triterpenoids in Saccharomyces cerevisiae by expression of CYP505D13 from Ganoderma lucidum

Xin Song,Wenfang Wang,Xiaozheng Wang,Han Xiao,Shangwen Luo,Shuangjun Lin

doi:10.1186/s40643-019-0256-6

Xin Song, Wenfang Wang + Show 4 more

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https://doi.org/10.1186/s40643-019-0256-6

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Abstract

BackgroundSqualene-type triterpenoids (STs) are a class of linearized triterpenoids with significant bioactivities, including anti-cancer, anti-oxidative, and anti-inflammatory activities. The efficient biosynthesis of STs has gained increasing attention.ResultsUsing Saccharomyces cerevisiae as a heterologous host, we discovered that overexpression of CYP505D13 from Ganoderma lucidum, a famous medicinal mushroom capable of producing various triterpenoids as secondary metabolites, enables the engineered S. cerevisiae strain to produce two new STs, 4,8-dihydroxy-22,23-oxidosqualene (ST-1), 8-hydroxy-2,3;22,23-squalene dioxide (ST-2), and a known ST, 2,3; 22,23-squalene dioxide (ST-3), at the respective titers of 3.28 mg/L, 13.77 mg/L, and 12.23 mg/L after 59 h fermentation. Furthermore, our in vitro enzymatic assay implies that CYP505D13 is involved in the formation of ST-3.ConclusionsThis study provides a promising alternative to discover STs and facilitate their efficient bioproduction.

Highlights

Squalene-type triterpenoids (STs) are a class of linearized triterpenoids with significant bioactivities, including anti-cancer, anti-oxidative, and anti-inflammatory activities
Along with the above mentioned paradigm, we discovered that overexpression of cyp505d13 from G. lucidum in S. cerevisiae YL-T3 yielded many compounds as compared to the control strain
Overexpression of CYP505D13 generates many new UPLC detectable peaks as compared to the control strain Since CYP505D13 from G. lucidum was co-expressed with lanosterol synthase (Chen et al 2012), it was initially considered as a candidate of lanosterol oxidase for biosynthesis of ganoderic acids, a group of lanostane-type triterpenoids (Wang et al 2018)

Summary

Results

Using Saccharomyces cerevisiae as a heterologous host, we discovered that overexpression of CYP505D13 from Ganoderma lucidum, a famous medicinal mushroom capable of producing various triterpenoids as secondary metabolites, enables the engineered S. cerevisiae strain to produce two new STs, 4,8-dihydroxy-22,23-oxidosqualene (ST-1), 8-hydroxy-2,3;22,23-squalene dioxide (ST-2), and a known ST, 2,3; 22,23-squalene dioxide (ST-3), at the respective titers of 3.28 mg/L, 13.77 mg/L, and 12.23 mg/L after 59 h fermentation. Our in vitro enzymatic assay implies that CYP505D13 is involved in the formation of ST-3

Background

Materials and methods

Results and discussion

Conclusion