Production of sesquiterpene patchoulol in mitochondrion-engineered Saccharomyces cerevisiae

Tao, Xinyi; Lin, Yang-Chen; Wang, Fengqing; Liu, Qinghai; Ma, Yushu; Liu, Min; Wei, Dongzhi

doi:10.1007/s10529-022-03240-3

“…183 Although the studies successfully produced their target compounds, the production came with a significant reduction in biomass accumulation attributed to the aforementioned toxicity of the pathway intermediates in the mitochondria. 181–183…”

Section: Targeting Localization Of Terpene Biosynthesis In Yeastmentioning

confidence: 99%

“…183 Although the studies successfully produced their target compounds, the production came with a signicant reduction in biomass accumulation attributed to the aforementioned toxicity of the pathway intermediates in the mitochondria. [181][182][183] Peroxisomes are small organelles involved in various functions such as energy metabolism. 178 In most cases, the organelles are not necessary for the viability of the cell, making it an attractive candidate for the production of terpenoids.…”

Section: Converting Yeast Organelles Into Terpenoid-producing Subcell...mentioning

confidence: 99%

“…182 The production of amorpha-4,11-diene and patchoulol was achieved by the localization of the amorpha-4,11-diene synthase (ADS) from Artemisia annua and patchoulol synthase from P. cablin. 183 Although the studies successfully produced their target compounds, the production came with a signicant reduction in biomass accumulation attributed to the aforementioned toxicity of the pathway intermediates in the mitochondria. [181][182][183] Peroxisomes are small organelles involved in various functions such as energy metabolism.…”

Section: Converting Yeast Organelles Into Terpenoid-producing Subcell...mentioning

confidence: 99%

See 1 more Smart Citation

Engineering yeast for the production of plant terpenoids using synthetic biology approaches

Bureau

¹

,

Oliva

²

,

Dong

³

et al. 2023

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“…[1] Thousands of sesquiterpenes have been identified, with numerous of them having significant commercial applications in drug [2] and fragrance development. [3] STPS-catalyzed reactions feature the carbocation-driven cascade cyclization, which is initiated by the departure of diphosphate from FPP to generate an allylic carbocation. The inherent reactivity of the carbocation enables a wide range of chemical reactions, including attacks by olefins, proton and hydride transfers, methyl and methylene migrations, and final quenching via deprotonation or hydration.…”

Section: Introductionmentioning

confidence: 99%

“…Sesquiterpene synthases (STPSs) contribute to the generation of at least 121 different sesquiterpene carbon skeletons from the universal substrate C 15 farnesyl diphosphate (FPP) [1] . Thousands of sesquiterpenes have been identified, with numerous of them having significant commercial applications in drug [2] and fragrance development [3] . STPS‐catalyzed reactions feature the carbocation‐driven cascade cyclization, which is initiated by the departure of diphosphate from FPP to generate an allylic carbocation.…”

Section: Introductionmentioning

confidence: 99%

Characterization and Engineering of Two Highly Paralogous Sesquiterpene Synthases Reveal a Regioselective Reprotonation Switch

Ye,

Shao,

Li

et al. 2024

Angewandte Chemie

0

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Sesquiterpene synthases (STPSs) catalyze carbocation‐driven cyclization reactions that can generate structurally diverse hydrocarbons. The deprotonation‐reprotonation process is widely used in STPSs to promote structural diversity, largely attributable to the distinct regio/stereoselective reprotonations. However, the molecular basis for reprotonation regioselectivity remains largely understudied. Herein, we analyzed two highly paralogous STPSs, Artabotrys hexapetalus(−)‐cyperene synthase (AhCS) and ishwarane synthase (AhIS), which catalyze reactions that are distinct from the regioselective protonation of germacrene A (GA), resulting in distinct skeletons of 5/5/6 tricyclic (−)‐cyperene and 6/6/5/3 tetracyclic ishwarane, respectively. Isotopic labeling experiments demonstrated that these protonations occur at C3 and C6 in AhCS and AhIS, respectively. The cryoelectron microscopy‐derived AhCS complex structure provided the structural basis for identifying different key active site residues that may govern their functional disparity. The structure‐guided mutagenesis of these residues resulted in successful functional interconversion between AhCS and AhIS, thus targeting the three active site residues [L311‐S419‐C458]/[M311‐V419‐A458] that may act as a C3/C6 reprotonation switch for GA. These findings could facilitate the rational design or directed evolution of STPSs with structurally diverse skeletons.

show abstract

Characterization and Engineering of Two Highly Paralogous Sesquiterpene Synthases Reveal a Regioselective Reprotonation Switch

Ye,

Shao,

Li

et al. 2024

Angew Chem Int Ed

1

0

View full text Add to dashboard Cite

Sesquiterpene synthases (STPSs) catalyze carbocation‐driven cyclization reactions that can generate structurally diverse hydrocarbons. The deprotonation‐reprotonation process is widely used in STPSs to promote structural diversity, largely attributable to the distinct regio/stereoselective reprotonations. However, the molecular basis for reprotonation regioselectivity remains largely understudied. Herein, we analyzed two highly paralogous STPSs, Artabotrys hexapetalus(−)‐cyperene synthase (AhCS) and ishwarane synthase (AhIS), which catalyze reactions that are distinct from the regioselective protonation of germacrene A (GA), resulting in distinct skeletons of 5/5/6 tricyclic (−)‐cyperene and 6/6/5/3 tetracyclic ishwarane, respectively. Isotopic labeling experiments demonstrated that these protonations occur at C3 and C6 in AhCS and AhIS, respectively. The cryoelectron microscopy‐derived AhCS complex structure provided the structural basis for identifying different key active site residues that may govern their functional disparity. The structure‐guided mutagenesis of these residues resulted in successful functional interconversion between AhCS and AhIS, thus targeting the three active site residues [L311‐S419‐C458]/[M311‐V419‐A458] that may act as a C3/C6 reprotonation switch for GA. These findings could facilitate the rational design or directed evolution of STPSs with structurally diverse skeletons.

show abstract

Production of sesquiterpene patchoulol in mitochondrion-engineered Saccharomyces cerevisiae

Cited by 10 publications

References 21 publications

Engineering yeast for the production of plant terpenoids using synthetic biology approaches

Engineering yeast for the production of plant terpenoids using synthetic biology approaches

Characterization and Engineering of Two Highly Paralogous Sesquiterpene Synthases Reveal a Regioselective Reprotonation Switch

Characterization and Engineering of Two Highly Paralogous Sesquiterpene Synthases Reveal a Regioselective Reprotonation Switch

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