Engineering chimeric diterpene synthases and isoprenoid biosynthetic pathways enables high-level production of miltiradiene in yeast

Hu, Tianyuan; Zhou, Jiawei; Tong, Yuru; Su, Ping; Li, Xinlin; Liu, Yuan; Liu, Nan; Wu, Xiao-Ning; Zhang, Yifeng; Wang, Jiadian; Gao, Linhui; Tu, Lichan; Lü, Yun; Jiang, Zhouqian; Zhou, Yongjin; Gao, Wei; Huang, Luqi

doi:10.1016/j.ymben.2020.03.011

Cited by 92 publications

(104 citation statements)

References 66 publications

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“…However, due to the use of antibiotics in the fermentation process to enhance the stability of the plasmid, it cannot be used in large-scale industrial production 114 . Recently, Tianyuan Hu et al investigated the production capacity of diterpenoid synthases from different species, and selected a class II diterpene synthase (di-TPS) Cf TPS1 from Coleus forskohlii ( Plectranthus barbatus ) and class I di-TPS Sm KSL1 from Salvia miltiorrhiza to use for producing a final titre of miltiradiene of 3.5 g/L in a 5-L bioreactor 115 .…”

Section: Biosynthesis Of Triptolidementioning

confidence: 99%

Triptolide: pharmacological spectrum, biosynthesis, chemical synthesis and derivatives

Gao¹,

Zhang²,

Liu³

et al. 2021

Theranostics

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Triptolide, an abietane-type diterpenoid isolated from Tripterygium wilfordii Hook. F., has significant pharmacological activity. Research results show that triptolide has obvious inhibitory effects on many solid tumors. Therefore, triptolide has become one of the lead compounds candidates for being the next "blockbuster" drug, and multiple triptolide derivatives have entered clinical research. An increasing number of researchers have developed triptolide synthesis methods to meet the clinical need. To provide new ideas for researchers in different disciplines and connect different disciplines with researchers aiming to solve scientific problems more efficiently, this article reviews the research progress made with analyzes of triptolide pharmacological activity, biosynthetic pathways, and chemical synthesis pathways and reported in toxicological and clinical studies of derivatives over the past 20 years, which have laid the foundation for subsequent researchers to study triptolide in many ways.

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Section: Biosynthesis Of Triptolidementioning

confidence: 99%

Triptolide: pharmacological spectrum, biosynthesis, chemical synthesis and derivatives

Gao¹,

Zhang²,

Liu³

et al. 2021

Theranostics

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“…The derived vectors were introduced into the indicated yeast strains, including BY4742 (MATα, his3Δ1, leu2Δ0, lys2Δ0, MET15, ura3Δ0) [42], BY-T1 (MATa, trp1Δ, his3Δ1, leu2Δ0, lys2Δ0, MET15, ura3Δ0, δDNA::P PGK1 -tHMG1-T ADH1 -P TEF1 -LYS2-T CYC1 ) [43], and BY-T20 (MATα, trp1Δ0, leu2Δ0, ura3Δ0,trp1::HIS3-P PGK1 -BTS1/ERG20-T ADH1 -P TDH3 -SaGGPS-T TPI1 -P TEF1 -tHMG1-T CYC1 ) [44] for required protein expression assays as well as CBT-ol and CBT-diol synthesis. And, the yeast transformants were selected by cultivation on SD (synthetic de ned) medium plates with desired dropout (DO) supplements (Takara, Japan).…”

Section: Vector Construction and Yeast Transformationmentioning

confidence: 99%

“…To develop microbes for tobacco cembranoid synthesis through the MVA pathway, the yeast strain BY4742 and its engineered strains BY-T1 and BT-T20 were utilized to develop the cembranoid synthetic systems. BY-T1 is a yeast strain expressing a truncated HMG-CoA reductase gene (tHMG1) to increase the upstream carbon ux to MVA pathway [43], and BY-T20 is a yeast strain with high e ciency GGPP production by expressing an engineered gene module composed of tHMG1, BTS1-ERG20 gene fusion and SaGGPS (GGPS from Sulfolobus acidocaldarius) [44]. Initially, pGADT7-CBTS1 vector was introduced into the yeast strain BY4742, BY-T1 and BY-T20 respectively for tobacco CBT-ol synthesis.…”

Section: Selection Of Yeast Strain For Cembranoid Synthesismentioning

confidence: 99%

Synthesis of cembratriene-ol and cembratriene-diol in yeast via the MVA pathway

Zhang

Bian

Liu

et al. 2021

Preprint

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Background: Cembranoids are one kind of diterpenoids with multiple biological activities. The tobacco cembratriene-ol (CBT-ol) and cembratriene-diol (CBT-diol) have high anti-insect and anti-fungal activities, which is attracting great attentions for their potential usage in sustainable agriculture. Cembranoids were supposed to be formed through the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway, yet the involvement of mevalonate (MVA) pathway in their synthesis remains unclear. Exploring the roles of MVA pathway in cembranoid synthesis could contribute not only to the technical approach but also to the molecular mechanism for cembranoid biosynthesis.Results: We constructed vectors to express cembratriene-ol synthase (CBTS1) and its fusion protein (AD-CBTS1) containing an N-terminal GAL4 AD domain as a translation leader in yeast. Eventually, the modified enzyme AD-CBTS1 was successfully expressed, which further resulted in the production of CBT-ol in the yeast strain BY-T20 with enhanced MVA pathway for geranylgeranyl diphosphate (GGPP) production but not in other yeast strains with low GGPP supply. Subsequently, CBT-diol was also synthesized by co-expression of the modified enzyme AD-CBTS1 and BD-CYP450 in the yeast strain BY-T20.Conclusions: We demonstrated that yeast is insensitive to the tobacco anti-fungal compound CBT-ol or CBT-diol and could be applied to their biosynthesis. This study further established a feasibility for cembranoid production via the MVA pathway and provided an alternative bio-approach for cembranoid biosynthesis in microbes.

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“…The lack of large amounts of celastrol present an obstacle in transforming celastrol into clinical therapeutics. Microbial cell factories, plant tissue culture, and total synthesis are effective strategies for obtaining target compounds, 209–211 which provide alternative approaches to obtain celastrol while preserving plant resources. The discovery of genes participating in the biosynthesis of celastrol and the development of synthetic biological strategies indicate the possibility of celastrol biosynthesis in heterologous hosts 20,21,63 .…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Biosynthesis, total synthesis, structural modifications, bioactivity, and mechanism of action of the quinone‐methide triterpenoid celastrol

Lü

Liu

Zhou

et al. 2020

Medicinal Research Reviews

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Celastrol, a quinone‐methide triterpenoid, was extracted from Tripterygium wilfordii Hook. F. in 1936 for the first time. Almost 70 years later, it is considered one of the molecules most likely to be developed into modern drugs, as it exhibits notable bioactivity, including anticancer and anti‐inflammatory activity, and exerts antiobesity effects. In addition, the molecular mechanisms underlying its bioactivity are being widely studied, which offers new avenues for its development as a pharmaceutical reagent. Owing to its potential therapeutic effects and unique chemical structure, celastrol has attracted considerable interest in the fields of organic, biosynthesis, and medicinal chemistry. As several steps in the biosynthesis of celastrol have been revealed, the mechanisms of key enzymes catalyzing the formation and postmodifications of the celastrol scaffold have been gradually elucidated, which lays a good foundation for the future heterogeneous biosynthesis of celastrol. Chemical synthesis is also an effective approach to obtain celastrol. The total synthesis of celastrol was realized for the first time in 2015, which established a new strategy to obtain celastroid natural products. However, owing to the toxic effects and suboptimal pharmacological properties of celastrol, its clinical applications remain limited. To search for drug‐like derivatives, several structurally modified compounds were synthesized and tested. This review focuses primarily on the latest research progress in the biosynthesis, total synthesis, structural modifications, bioactivity, and mechanism of action of celastrol. We anticipate that this paper will facilitate a more comprehensive understanding of this promising compound and provide constructive references for future research in this field.

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Engineering chimeric diterpene synthases and isoprenoid biosynthetic pathways enables high-level production of miltiradiene in yeast

Cited by 92 publications

References 66 publications

Triptolide: pharmacological spectrum, biosynthesis, chemical synthesis and derivatives

Triptolide: pharmacological spectrum, biosynthesis, chemical synthesis and derivatives

Synthesis of cembratriene-ol and cembratriene-diol in yeast via the MVA pathway

Biosynthesis, total synthesis, structural modifications, bioactivity, and mechanism of action of the quinone‐methide triterpenoid celastrol

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