Dear Editor,Plant UDP-dependent glycosyltransferases(UGTs),belonging to the carbohydrate-active enzyme glycosyltransferase 1 family(Louveau and Osbourn,2019),not only play important roles in adaptation to various envi...Dear Editor,Plant UDP-dependent glycosyltransferases(UGTs),belonging to the carbohydrate-active enzyme glycosyltransferase 1 family(Louveau and Osbourn,2019),not only play important roles in adaptation to various environments(Cai et al.,2020;Pastorczyk-Szlenkier and Bednarek,2021)but also endow plant natural products with great pharmaceutical and ecological significance(Margolin et al.,2020).In recent years,an increasing number of plant UGTs have been characterized to function in the biosynthesis of many bioactive compounds such as ginsenosides(Wei et al.,2015),breviscapine(Liu et al.,2018),and rubusoside(Xu et al.,2022).展开更多
Plant natural products(PNPs)are the main sources of drugs,food additives,and new biofuels and have become a hotspot in synthetic biology.In the past two decades,the engineered biosynthesis of many PNPs has been achiev...Plant natural products(PNPs)are the main sources of drugs,food additives,and new biofuels and have become a hotspot in synthetic biology.In the past two decades,the engineered biosynthesis of many PNPs has been achieved through the construction of microbial cell factories.Alongside the rapid development of plant physiology,genetics,and plant genetic modification techniques,hosts have now expanded from single-celled microbes to complex plant systems.Plant synthetic biology is an emerging field that combines engineering principles with plant biology.In this review,we introduce recent advances in the biosynthetic pathway elucidation of PNPs and summarize the progress of engineered PNP biosynthesis in plant cells.Furthermore,a future vision of plant synthetic biology is proposed.Although we are still a long way from overcoming all the bottlenecks in plant synthetic biology,the ascent of this field is expected to provide a huge opportunity for future agriculture and industry.展开更多
Monoterpenoids are typically present in the secretory tissues of higher plants,and their biosynthesis is catalyzed by the action of monoterpene synthases(MTSs).However,the knowledge about these enzymes is restricted i...Monoterpenoids are typically present in the secretory tissues of higher plants,and their biosynthesis is catalyzed by the action of monoterpene synthases(MTSs).However,the knowledge about these enzymes is restricted in a few plant species.MTSs are responsible for the complex cyclization of monoterpene precursors,resulting in the production of diverse monoterpene products.These enzymatic reactions are considered exceptionally complex in nature.Therefore,it is crucial to understand the catalytic mechanism of MTSs to elucidate their ability to produce diverse or specific monoterpenoid products.In our study,we analyzed thirteen genomes of Dipterocarpaceae and identified 38 MTSs that generate a variety of monoterpene products.By focusing on four MTSs with different product spectra and analyzing the formation mechanism of acyclic,monocyclic and bicyclic products in MTSs,we observed that even a single amino acid mutation can change the specificity and diversity of MTS products,which is due to the synergistic effect between the shape of the active cavity and the stabilization of carbon-positive intermediates that the mutation changing.Notably,residues N340,I448,and phosphoric acid groups were found to be significant contributors to the stabilization of intermediate terpinyl and pinene cations.Alterations in these residues,either directly or indirectly,can impact the synthesis of single monoterpenes or their mixtures.By revealing the role of key residues in the catalytic process and establishing the interaction model between specific residues and complex monoterpenes in MTSs,it will be possible to reasonably design and engineer different catalytic activities into existing MTSs,laying a foundation for the artificial design and industrial application of MTSs.展开更多
Taxanes are kinds of diterpenoids with important bioactivities,such as paclitaxel(taxol®)is an excellent natural broad-spectrum anticancer drug.Attempts to biosynthesize taxanes have made with limited success,mai...Taxanes are kinds of diterpenoids with important bioactivities,such as paclitaxel(taxol®)is an excellent natural broad-spectrum anticancer drug.Attempts to biosynthesize taxanes have made with limited success,mainly due to the bottleneck of the low efficiency catalytic elements.In this study,we developed an artificial synthetic system to produce taxanes from mevalonate(MVA)by coupling biological and chemical methods,which comprises in vitro multi-enzyme catalytic module,chemical catalytic module and yeast cell catalytic module.Through optimizing in vitro multienzyme catalytic system,the yield of taxadiene was increased to 946.7 mg/L from MVA within 8 h and the productivity was 14.2-fold higher than microbial fermentation.By incorporating palladium catalysis,the conversion rate of Taxa-4(20),11(12)-dien-5α-yl acetate(T5α-AC)reached 48%,effectively addressing the product promiscuity and the low yield rate of T5αOH.Finally,we optimized the expression of T10βOH in yeast resulting in the biosynthesis of Taxa-4(20),11(12)-dien-5α-acetoxy-10β-ol(T5α-AC-10β-ol)at a production of 15.8 mg/L,which displayed more than 2000-fold higher than that produced by co-culture fermentation strategy.These technologies offered a promising new approach for efficient synthesis of taxanes.展开更多
基金supported by grants from the National Key R&D Program of China(no.2019YFA0905700)Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project(TSBICIP-CXRC-015)+1 种基金China Postdoctoral Science Foundation(No.2019M661032)National Natural Science Foundation of China(No.31901026).
文摘Dear Editor,Plant UDP-dependent glycosyltransferases(UGTs),belonging to the carbohydrate-active enzyme glycosyltransferase 1 family(Louveau and Osbourn,2019),not only play important roles in adaptation to various environments(Cai et al.,2020;Pastorczyk-Szlenkier and Bednarek,2021)but also endow plant natural products with great pharmaceutical and ecological significance(Margolin et al.,2020).In recent years,an increasing number of plant UGTs have been characterized to function in the biosynthesis of many bioactive compounds such as ginsenosides(Wei et al.,2015),breviscapine(Liu et al.,2018),and rubusoside(Xu et al.,2022).
基金supported by grants from the National Natural Science Foundation of China(grant no.31901026)the China Postdoctoral Science Foundation(grant no.2019M661032)Tianjin Science and technology plan project(grant no.19PTZWHZ00060).
文摘Plant natural products(PNPs)are the main sources of drugs,food additives,and new biofuels and have become a hotspot in synthetic biology.In the past two decades,the engineered biosynthesis of many PNPs has been achieved through the construction of microbial cell factories.Alongside the rapid development of plant physiology,genetics,and plant genetic modification techniques,hosts have now expanded from single-celled microbes to complex plant systems.Plant synthetic biology is an emerging field that combines engineering principles with plant biology.In this review,we introduce recent advances in the biosynthetic pathway elucidation of PNPs and summarize the progress of engineered PNP biosynthesis in plant cells.Furthermore,a future vision of plant synthetic biology is proposed.Although we are still a long way from overcoming all the bottlenecks in plant synthetic biology,the ascent of this field is expected to provide a huge opportunity for future agriculture and industry.
基金supported by the National Key R&D Program of China(2020YFA0908000)the National Natural Science Foundation of China(31901015)Science and Technology Partnership Program,Ministry of Science and Technology of China(KY202001017).
文摘Monoterpenoids are typically present in the secretory tissues of higher plants,and their biosynthesis is catalyzed by the action of monoterpene synthases(MTSs).However,the knowledge about these enzymes is restricted in a few plant species.MTSs are responsible for the complex cyclization of monoterpene precursors,resulting in the production of diverse monoterpene products.These enzymatic reactions are considered exceptionally complex in nature.Therefore,it is crucial to understand the catalytic mechanism of MTSs to elucidate their ability to produce diverse or specific monoterpenoid products.In our study,we analyzed thirteen genomes of Dipterocarpaceae and identified 38 MTSs that generate a variety of monoterpene products.By focusing on four MTSs with different product spectra and analyzing the formation mechanism of acyclic,monocyclic and bicyclic products in MTSs,we observed that even a single amino acid mutation can change the specificity and diversity of MTS products,which is due to the synergistic effect between the shape of the active cavity and the stabilization of carbon-positive intermediates that the mutation changing.Notably,residues N340,I448,and phosphoric acid groups were found to be significant contributors to the stabilization of intermediate terpinyl and pinene cations.Alterations in these residues,either directly or indirectly,can impact the synthesis of single monoterpenes or their mixtures.By revealing the role of key residues in the catalytic process and establishing the interaction model between specific residues and complex monoterpenes in MTSs,it will be possible to reasonably design and engineer different catalytic activities into existing MTSs,laying a foundation for the artificial design and industrial application of MTSs.
基金supported in part by the National Natural Science Foundation of China(32371499,31901026 and 32172443),China Postdoctoral Science Foundation(2019M661032)National Key R&D Program of China(2020YFA0908000 and 2022YFD1700400)+1 种基金the Frontiers Science Center for New Organic Matter,Nankai University(63181206)he Tianjin Development Program for Innovation and Entrepreneurship(2019-06).
文摘Taxanes are kinds of diterpenoids with important bioactivities,such as paclitaxel(taxol®)is an excellent natural broad-spectrum anticancer drug.Attempts to biosynthesize taxanes have made with limited success,mainly due to the bottleneck of the low efficiency catalytic elements.In this study,we developed an artificial synthetic system to produce taxanes from mevalonate(MVA)by coupling biological and chemical methods,which comprises in vitro multi-enzyme catalytic module,chemical catalytic module and yeast cell catalytic module.Through optimizing in vitro multienzyme catalytic system,the yield of taxadiene was increased to 946.7 mg/L from MVA within 8 h and the productivity was 14.2-fold higher than microbial fermentation.By incorporating palladium catalysis,the conversion rate of Taxa-4(20),11(12)-dien-5α-yl acetate(T5α-AC)reached 48%,effectively addressing the product promiscuity and the low yield rate of T5αOH.Finally,we optimized the expression of T10βOH in yeast resulting in the biosynthesis of Taxa-4(20),11(12)-dien-5α-acetoxy-10β-ol(T5α-AC-10β-ol)at a production of 15.8 mg/L,which displayed more than 2000-fold higher than that produced by co-culture fermentation strategy.These technologies offered a promising new approach for efficient synthesis of taxanes.
