Electrocatalytic water splitting for hydrogen production is an appealing strategy to reduce carbon emissions and generate renewable fuels.This promising process,however,is limited by its sluggish reaction kinetics and...Electrocatalytic water splitting for hydrogen production is an appealing strategy to reduce carbon emissions and generate renewable fuels.This promising process,however,is limited by its sluggish reaction kinetics and high-cost catalysts.The two-dimensional(2D)transition metal dichalcogenides(TMDCs)have presented great potential as electrocatalytic materials due to their tunable bandgaps,abundant defective active sites,and good chemical stability.Consequently,phase engineering,defect engineering and interface engineering have been adopted to manipulate the electronic structure of TMDCs for boosting their exceptional catalytic performance.Particularly,it is essential to clarify the local structure of catalytically active sites of TMDCs and their structural evolution in catalytic reactions using atomic resolution electron microscopy and the booming in situ technologies,which is beneficial for exploring the underlying reaction mechanism.In this review,the growth regulation,characterization,particularly atomic configurations of active sites in TMDCs are summarized.The significant role of electron microscopy in the understanding of the growth mechanism,the controlled synthesis and functional optimization of 2D TMDCs are discussed.This review will shed light on the design and synthesis of novel electrocatalysts with high performance,as well as prompt the application of advanced electron microscopy in the research of materials science.展开更多
During the process of designing the mixed-flow pump impeller, the meridional flow passage shape directly affects the obtained meridional flow field, which then has an influence on the three-dimensional impeller shape....During the process of designing the mixed-flow pump impeller, the meridional flow passage shape directly affects the obtained meridional flow field, which then has an influence on the three-dimensional impeller shape. However, the meridional flow passage shape is too complicated to be described by a simple formula for now. Therefore, reasonable parameter selection for the meridional flow passage is essential to the investigation. In order to explore the effects of the meridional flow passage shape on the impeller design and the hydraulic performance of the mixed-flow pump, the hub and shroud radius ratio (HSRR) of impeller and the outlet diffusion angle (ODA) of outlet zone are selected as the meridional flow passage parameters. 25 mixed-flow pump impellers, with specific speed of 496 under the design condition, are designed with various parameter combinations. Among these impellers, one with HSRR of 1.94 and ODA of 90° is selected to carry out the model test and the obtained experimental results are used to verify accuracies of the head and the hydraulic efficiency predicted by numerical simulation. Based on SIMPLE algorithm and standard k-ε two-equation turbulence model, the three-dimensional steady incompressible Reynolds averaged Navier-Stokes equations are solved and the effects of different parameters on hydraulic performance of mixed-flow pump impellers are analyzed. The analysis results demonstrate that there are optimal values of HSRR and ODA available, so the hydraulic performance and the internal flow of mixed-flow pumps can be improved by selecting appropriate values for the meridional flow passage parameters. The research on these two parameters, HSRR and ODA, has further illustrated influences of the meridional flow passage shape on the hydraulic performance of the mixed-flow pump, and is beneficial to improving the design of the mixed-flow pump impeller.展开更多
The existing research on improving the hydraulic performance of centrifugal pumps mainly focuses on the design method and the parameter optimization. The traditional design method for centrifugal impellers relies more...The existing research on improving the hydraulic performance of centrifugal pumps mainly focuses on the design method and the parameter optimization. The traditional design method for centrifugal impellers relies more on experience of engineers that typically only satisfies the continuity equation of the fluid. In this study, on the basis of the direct and inverse iteration design method which simultaneously solves the continuity and motion equations of the fluid and shapes the blade geometry by controlling the wrap angle, three centrifugal pump impellers are designed by altering blade wrap angles while keeping other parameters constant. The three-dimensional flow fields in three centrifugal pumps are numerically simulated, and the simulation results illustrate that the blade with larger wrap angle has more powerful control ability on the flow pattern in impeller. The three pumps have nearly the same pressure distributions at the small flow rate, but the pressure gradient increase in the pump with the largest wrap angle is smoother than the other two pumps at the design and large flow rates. The pump head and efficiency are also influenced by the blade wrap angle. The highest head and efficiency are also observed for the largest angle. An experiment rig is designed and built to test the performance of the pump with the largest wrap angle. The test results show that the wide space of its efficiency area and the stability of its operation ensure the excellent performance of the design method and verify the numerical analysis. The analysis on influence of the blade wrap angle for centrifugal pump performance in this paper can be beneficial to the optimization design of the centrifugal pump.展开更多
In the mixed-flow pump design, the shape of the flow passage can directly affect the flow capacity and the internal flow, thus influencing hydraulic performance, cavitation performance and operation stability of the m...In the mixed-flow pump design, the shape of the flow passage can directly affect the flow capacity and the internal flow, thus influencing hydraulic performance, cavitation performance and operation stability of the mixed-flow pump. However, there is currently a lack of experimental research on the influence mechanism. Therefore, in order to analyze the effects of subtle variations of the flow passage on the mixed-flow pump performance, the frustum cone surface of the end part of inlet contraction flow passage of the mixed-flow pump is processed into a cylindrical surface and a test rig is built to carry out the hydraulic performance experiment. In this experiment, parameters, such as the head, the efficiency, and the shaft power, are measured, and the pressure fluctuation and the noise signal are also collected. The research results suggest that after processing the inlet flow passage, the head of the mixed-flow pump significantly goes down; the best efficiency of the mixed-flow pump drops by approximately 1.5%, the efficiency decreases more significantly under the large flow rate; the shaft power slightly increases under the large flow rate, slightly decreases under the small flow rate. In addition, the pressure fluctuation amplitudes on both the impeller inlet and the diffuser outlet increase significantly with more drastic pressure fluctuations and significantly lower stability of the internal flow of the mixed-flow pump. At the same time, the noise dramatically increases. Overall speaking, the subtle variation of the inlet flow passage leads to a significant change of the mixed-flow pump performance, thus suggesting a special attention to the optimization of flow passage. This paper investigates the influence of the flow passage variation on the mixed-flow pump performance by experiment, which will benefit the optimal design of the flow passage of the mixed-flow pump.展开更多
The effects of trace yttrium(Y)element on the microstructure,mechanical properties,and corrosion resistance of Mg-2Zn-0.1Mn-0.3Ca-xY(x=0,0.1,0.2,0.3)biological magnesium alloys are investigated.Results show that grain...The effects of trace yttrium(Y)element on the microstructure,mechanical properties,and corrosion resistance of Mg-2Zn-0.1Mn-0.3Ca-xY(x=0,0.1,0.2,0.3)biological magnesium alloys are investigated.Results show that grain size decreases from 310 to 144µm when Y content increases from 0wt%to 0.3wt%.At the same time,volume fraction of the second phase increases from 0.4%to 6.0%,yield strength of the alloy continues to increase,and ultimate tensile strength and elongation decrease initially and then increase.When the Y content increases to 0.3wt%,Mg_(3)Zn_(6)Y phase begins to precipitate in the alloy;thus,the alloy exhibits the most excellent mechanical property.At this time,its ultimate tensile strength,yield strength,and elongation are 119 MPa,69 MPa,and 9.1%,respectively.In addition,when the Y content is 0.3wt%,the alloy shows the best corrosion resistance in the simulated body fluid(SBF).This investigation has revealed that the improvement of mechanical properties and corrosion resistance is mainly attributed to the grain refinement and the precipitated Mg_(3)Zn_(6)Y phase.展开更多
Spontaneous rupture of hepatocellular carcinoma(HCC) is a life-threatening complication and its prognosis is significantly poor because of the high recurrence rate after initial hepatectomy. Resection of isolated extr...Spontaneous rupture of hepatocellular carcinoma(HCC) is a life-threatening complication and its prognosis is significantly poor because of the high recurrence rate after initial hepatectomy. Resection of isolated extrahepatic metastasis of HCC has been advocated to obtain a possibility of long-term survival. However, it is a challenge for clinicians to detect implantation metastasis of spontaneously ruptured HCC. Accurate re-staging plays the most important role in making a decision on isolated metastasis resection. 18F-fluorodeoxyglucose(18F-FDG) positron emission tomography/computed tomography(PET/CT) is useful in detecting intraabdominal implantation metastasis from a variety of malignancies and shows superior accuracy to conventional imaging modalities in determining the location of metastasis. We present one patient with a new isolated pelvic implantation metastasis detected by 18F-FDG PET/CT and pathologically confirmed by PET/CT-guided percutaneous biopsy, who had a history of resection of spontaneously ruptured HCC two years ago. The patient's condition was stable at the 6-mo follow-up after resection of the isolated pelvic metastasis.展开更多
Objective: To study the evaluation value of three-dimensional finite element model analysis for bone mineral density (BMD) and bone metabolism activity in patients with osteoporosis. Methods: A total of 218 patients w...Objective: To study the evaluation value of three-dimensional finite element model analysis for bone mineral density (BMD) and bone metabolism activity in patients with osteoporosis. Methods: A total of 218 patients who were diagnosed with osteoporosis in the hospital between February 2014 and January 2017 were collected as observation group, and 100 healthy volunteers who received physical examination in the hospital during the same period were selected as normal control group. The femoral head of the two groups was analyzed by three-dimensional finite element model, and the femoral head BMD levels and serum bone metabolism index contents were measured. Pearson test was used to evaluate the evaluation value of femoral head three-dimensional finite element model for osteoporosis. Results: The cancellous bone and cortical bone Von Mises stress value of observation group were lower than those of normal control group, and femoral neck BMD value of observation group was lower than that of normal control group;serum bone metabolism index BGP content was lower than that of normal control group while NBAP, TRACP-5b and CTX-1 contents were higher than those of normal control group. Pearson test showed that the cancellous bone and cortical bone Von Mises stress value of patients with osteoporosis were directly correlated with BMD value and bone metabolism index contents. Conclusion: The three-dimensional finite element model analysis resultsof patients with osteoporosis can objectively reflect the femoral headBMD value and bone metabolism activity, and is a reliable way to evaluate the risk of long-term fractures.展开更多
Background Chemotherapy can have a negative impact on the growth of children with different cancers.However,few studies have examined whether intravenous chemotherapy(IVC)affects the growth of children with retinoblas...Background Chemotherapy can have a negative impact on the growth of children with different cancers.However,few studies have examined whether intravenous chemotherapy(IVC)affects the growth of children with retinoblastoma(RB).The present study evaluated the height differences(actual height compared to the age standardized value)and survival of pediatric RB patients treated with IVC.Methods This was an observational cohort study.A total of 87 pediatric RB patients were included.The study population was stratified into two groups based on the number of chemotherapy cycles administered(≤4 versus>4).The height at baseline(before IVC),height after IVC and overall survival were compared between the two groups.Results Before IVC,no height differences were observed between the two groups(P=0.585).After IVC,all of the patients had a reduced height compared to the age standardized height(P=0.035).Patients who underwent more cycles of chemotherapy had a greater height difference compared to those who received fewer cycles(P=0.008).For those who had reduced height,the difference was positively associated with the number of chemotherapy cycles(r=0.279,P=0.043).Among the patients who exhibited a greater height difference,those who underwent more than four cycles of chemotherapy had a decreased overall survival(P=0.042).Conclusions Pediatric RB patients who underwent more cycles of chemotherapy were more likely to have a reduced height.Further studies are needed to determine the optimal treatment strategy to prevent the reduced growth while maintaining the benefits of chemotherapy.展开更多
Cucurbitacin IIa is a triterpenoid isolated exclusively from Hemsleya plants and a non-steroidal anti-inflammatory drug that functions as the main ingredient of prescription Hemslecin capsules and tablets in China.Syn...Cucurbitacin IIa is a triterpenoid isolated exclusively from Hemsleya plants and a non-steroidal anti-inflammatory drug that functions as the main ingredient of prescription Hemslecin capsules and tablets in China.Synthetic biology provides new strategies for production of such valuable cucurbitacins at a large scale;however,the biosynthetic pathway of cucurbitacin IIa has been unknown,and the heterologous production of cucurbitacins in galactose medium has been expensive and low yielding.In this study,we characterized the functions of genes encoding two squalene epoxidases(HcSE1-2),six oxidosqualene cyclases(HcOSC1-6),two CYP450s(HcCYP87D20 and HcCYP81Q59),and an acyltransferase(HcAT1)in cucurbitacin IIa biosynthesis by heterologous expression in Saccharomyces cerevisiae and Nicotiana benthamiana.We achieved high-level production of the key cucurbitacin precursor 11-carbonyl-20b-hydroxy-Cuol from glucose in yeast via modular engineering of the mevalonate pathway and optimization of P450 expression levels.The resulting yields of 46.41 mg/l 11-carbonyl-20b-hydroxy-Cuol and 126.47 mg/l total cucurbitacin triterpenoids in shake flasks are the highest yields yet reported from engineered microbes.Subsequently,production of 11-carbonyl-20b-hydroxy-Cuol by transient gene expression in tobacco resulted in yields of 1.28 mg/g dry weight in leaves.This work reveals the key genes involved in biosynthesis of prescription cucurbitacin IIa and demonstrates that engineered yeast cultivated with glucose can produce high yields of key triterpenoid intermediates.We describe a low-cost and highly efficient platform for rapid screening of candidate genes and high-yield production of pharmacological triterpenoids.展开更多
基金the National Natural Science Foundation of China(Grant Nos.U21A20174 and 52001222)the Science and Technology Innovation Talent Team Project of Shanxi Province(Grant No.202304051001010)+3 种基金the Key National Scientific and Technological Co-operation Projects of Shanxi Province(Grant No.202104041101008)the Natural Science Foundation of Shanxi Province(Grant No.202303021221045)the Program for the Innovative Talents of Higher Education Institutions of Shanxi(PTIT)and the Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi(STIP)(Grant No.2022L036).
文摘Electrocatalytic water splitting for hydrogen production is an appealing strategy to reduce carbon emissions and generate renewable fuels.This promising process,however,is limited by its sluggish reaction kinetics and high-cost catalysts.The two-dimensional(2D)transition metal dichalcogenides(TMDCs)have presented great potential as electrocatalytic materials due to their tunable bandgaps,abundant defective active sites,and good chemical stability.Consequently,phase engineering,defect engineering and interface engineering have been adopted to manipulate the electronic structure of TMDCs for boosting their exceptional catalytic performance.Particularly,it is essential to clarify the local structure of catalytically active sites of TMDCs and their structural evolution in catalytic reactions using atomic resolution electron microscopy and the booming in situ technologies,which is beneficial for exploring the underlying reaction mechanism.In this review,the growth regulation,characterization,particularly atomic configurations of active sites in TMDCs are summarized.The significant role of electron microscopy in the understanding of the growth mechanism,the controlled synthesis and functional optimization of 2D TMDCs are discussed.This review will shed light on the design and synthesis of novel electrocatalysts with high performance,as well as prompt the application of advanced electron microscopy in the research of materials science.
基金supported by National Natural Science Foundation of China (Grant No. 51176088)
文摘During the process of designing the mixed-flow pump impeller, the meridional flow passage shape directly affects the obtained meridional flow field, which then has an influence on the three-dimensional impeller shape. However, the meridional flow passage shape is too complicated to be described by a simple formula for now. Therefore, reasonable parameter selection for the meridional flow passage is essential to the investigation. In order to explore the effects of the meridional flow passage shape on the impeller design and the hydraulic performance of the mixed-flow pump, the hub and shroud radius ratio (HSRR) of impeller and the outlet diffusion angle (ODA) of outlet zone are selected as the meridional flow passage parameters. 25 mixed-flow pump impellers, with specific speed of 496 under the design condition, are designed with various parameter combinations. Among these impellers, one with HSRR of 1.94 and ODA of 90° is selected to carry out the model test and the obtained experimental results are used to verify accuracies of the head and the hydraulic efficiency predicted by numerical simulation. Based on SIMPLE algorithm and standard k-ε two-equation turbulence model, the three-dimensional steady incompressible Reynolds averaged Navier-Stokes equations are solved and the effects of different parameters on hydraulic performance of mixed-flow pump impellers are analyzed. The analysis results demonstrate that there are optimal values of HSRR and ODA available, so the hydraulic performance and the internal flow of mixed-flow pumps can be improved by selecting appropriate values for the meridional flow passage parameters. The research on these two parameters, HSRR and ODA, has further illustrated influences of the meridional flow passage shape on the hydraulic performance of the mixed-flow pump, and is beneficial to improving the design of the mixed-flow pump impeller.
基金supported by National Natural Science Foundation of China(Grant Nos.51176088,51179090)National Basic Research Program of China(973 Program,Grant No.2009CB724304)+1 种基金General Financial Grant from the China Postdoctoral Science Foundation(Grant No.2011M500315)Open Research Fund Program of State Key Laboratory of Hydroscience and Engineering of China(Grant No.sklhse-2012-E-02)
文摘The existing research on improving the hydraulic performance of centrifugal pumps mainly focuses on the design method and the parameter optimization. The traditional design method for centrifugal impellers relies more on experience of engineers that typically only satisfies the continuity equation of the fluid. In this study, on the basis of the direct and inverse iteration design method which simultaneously solves the continuity and motion equations of the fluid and shapes the blade geometry by controlling the wrap angle, three centrifugal pump impellers are designed by altering blade wrap angles while keeping other parameters constant. The three-dimensional flow fields in three centrifugal pumps are numerically simulated, and the simulation results illustrate that the blade with larger wrap angle has more powerful control ability on the flow pattern in impeller. The three pumps have nearly the same pressure distributions at the small flow rate, but the pressure gradient increase in the pump with the largest wrap angle is smoother than the other two pumps at the design and large flow rates. The pump head and efficiency are also influenced by the blade wrap angle. The highest head and efficiency are also observed for the largest angle. An experiment rig is designed and built to test the performance of the pump with the largest wrap angle. The test results show that the wide space of its efficiency area and the stability of its operation ensure the excellent performance of the design method and verify the numerical analysis. The analysis on influence of the blade wrap angle for centrifugal pump performance in this paper can be beneficial to the optimization design of the centrifugal pump.
基金Supported by National Natural Science Foundation of China(Grant No.51176088)
文摘In the mixed-flow pump design, the shape of the flow passage can directly affect the flow capacity and the internal flow, thus influencing hydraulic performance, cavitation performance and operation stability of the mixed-flow pump. However, there is currently a lack of experimental research on the influence mechanism. Therefore, in order to analyze the effects of subtle variations of the flow passage on the mixed-flow pump performance, the frustum cone surface of the end part of inlet contraction flow passage of the mixed-flow pump is processed into a cylindrical surface and a test rig is built to carry out the hydraulic performance experiment. In this experiment, parameters, such as the head, the efficiency, and the shaft power, are measured, and the pressure fluctuation and the noise signal are also collected. The research results suggest that after processing the inlet flow passage, the head of the mixed-flow pump significantly goes down; the best efficiency of the mixed-flow pump drops by approximately 1.5%, the efficiency decreases more significantly under the large flow rate; the shaft power slightly increases under the large flow rate, slightly decreases under the small flow rate. In addition, the pressure fluctuation amplitudes on both the impeller inlet and the diffuser outlet increase significantly with more drastic pressure fluctuations and significantly lower stability of the internal flow of the mixed-flow pump. At the same time, the noise dramatically increases. Overall speaking, the subtle variation of the inlet flow passage leads to a significant change of the mixed-flow pump performance, thus suggesting a special attention to the optimization of flow passage. This paper investigates the influence of the flow passage variation on the mixed-flow pump performance by experiment, which will benefit the optimal design of the flow passage of the mixed-flow pump.
基金financially supported by the National Natural Science Foundation of China(Nos.52005034 and 52027805)the China Postdoctoral Science Foundation Funded Project(No.2021M691860)+2 种基金the Beijing Postdoctoral Research Foundation(No.2021-ZZ-073)the Zhuhai Industry-University-Research Cooperation Project(No.ZH22017001200176PWC)the Tai’an City Science and Technology Innovation Major Project(No.2021ZDZX011)。
文摘The effects of trace yttrium(Y)element on the microstructure,mechanical properties,and corrosion resistance of Mg-2Zn-0.1Mn-0.3Ca-xY(x=0,0.1,0.2,0.3)biological magnesium alloys are investigated.Results show that grain size decreases from 310 to 144µm when Y content increases from 0wt%to 0.3wt%.At the same time,volume fraction of the second phase increases from 0.4%to 6.0%,yield strength of the alloy continues to increase,and ultimate tensile strength and elongation decrease initially and then increase.When the Y content increases to 0.3wt%,Mg_(3)Zn_(6)Y phase begins to precipitate in the alloy;thus,the alloy exhibits the most excellent mechanical property.At this time,its ultimate tensile strength,yield strength,and elongation are 119 MPa,69 MPa,and 9.1%,respectively.In addition,when the Y content is 0.3wt%,the alloy shows the best corrosion resistance in the simulated body fluid(SBF).This investigation has revealed that the improvement of mechanical properties and corrosion resistance is mainly attributed to the grain refinement and the precipitated Mg_(3)Zn_(6)Y phase.
基金Supported by National Science Foundation for Yong Scholars of China,No.81101067
文摘Spontaneous rupture of hepatocellular carcinoma(HCC) is a life-threatening complication and its prognosis is significantly poor because of the high recurrence rate after initial hepatectomy. Resection of isolated extrahepatic metastasis of HCC has been advocated to obtain a possibility of long-term survival. However, it is a challenge for clinicians to detect implantation metastasis of spontaneously ruptured HCC. Accurate re-staging plays the most important role in making a decision on isolated metastasis resection. 18F-fluorodeoxyglucose(18F-FDG) positron emission tomography/computed tomography(PET/CT) is useful in detecting intraabdominal implantation metastasis from a variety of malignancies and shows superior accuracy to conventional imaging modalities in determining the location of metastasis. We present one patient with a new isolated pelvic implantation metastasis detected by 18F-FDG PET/CT and pathologically confirmed by PET/CT-guided percutaneous biopsy, who had a history of resection of spontaneously ruptured HCC two years ago. The patient's condition was stable at the 6-mo follow-up after resection of the isolated pelvic metastasis.
基金National Science Foundation of China No:81301292.
文摘Objective: To study the evaluation value of three-dimensional finite element model analysis for bone mineral density (BMD) and bone metabolism activity in patients with osteoporosis. Methods: A total of 218 patients who were diagnosed with osteoporosis in the hospital between February 2014 and January 2017 were collected as observation group, and 100 healthy volunteers who received physical examination in the hospital during the same period were selected as normal control group. The femoral head of the two groups was analyzed by three-dimensional finite element model, and the femoral head BMD levels and serum bone metabolism index contents were measured. Pearson test was used to evaluate the evaluation value of femoral head three-dimensional finite element model for osteoporosis. Results: The cancellous bone and cortical bone Von Mises stress value of observation group were lower than those of normal control group, and femoral neck BMD value of observation group was lower than that of normal control group;serum bone metabolism index BGP content was lower than that of normal control group while NBAP, TRACP-5b and CTX-1 contents were higher than those of normal control group. Pearson test showed that the cancellous bone and cortical bone Von Mises stress value of patients with osteoporosis were directly correlated with BMD value and bone metabolism index contents. Conclusion: The three-dimensional finite element model analysis resultsof patients with osteoporosis can objectively reflect the femoral headBMD value and bone metabolism activity, and is a reliable way to evaluate the risk of long-term fractures.
基金supported by the Special Fund of Science and Technology Innovation for Social Livelihood in Chongqing(no.cstc2015shmszxl20107,Hongfeng Yuan).
文摘Background Chemotherapy can have a negative impact on the growth of children with different cancers.However,few studies have examined whether intravenous chemotherapy(IVC)affects the growth of children with retinoblastoma(RB).The present study evaluated the height differences(actual height compared to the age standardized value)and survival of pediatric RB patients treated with IVC.Methods This was an observational cohort study.A total of 87 pediatric RB patients were included.The study population was stratified into two groups based on the number of chemotherapy cycles administered(≤4 versus>4).The height at baseline(before IVC),height after IVC and overall survival were compared between the two groups.Results Before IVC,no height differences were observed between the two groups(P=0.585).After IVC,all of the patients had a reduced height compared to the age standardized height(P=0.035).Patients who underwent more cycles of chemotherapy had a greater height difference compared to those who received fewer cycles(P=0.008).For those who had reduced height,the difference was positively associated with the number of chemotherapy cycles(r=0.279,P=0.043).Among the patients who exhibited a greater height difference,those who underwent more than four cycles of chemotherapy had a decreased overall survival(P=0.042).Conclusions Pediatric RB patients who underwent more cycles of chemotherapy were more likely to have a reduced height.Further studies are needed to determine the optimal treatment strategy to prevent the reduced growth while maintaining the benefits of chemotherapy.
基金supported by the Major Science and Technology Projects in Yunnan Province(2019ZF011-1)the Fundamental Research Project of Yunnan(202101AS070037)+4 种基金the Science and Technology Innovation team of Yunnan(202105AE160011)the Major Science and Technique Programs in Yunnan Province(202102AE090042)the Yunnan Characteristic Plant Extraction Laboratory(2022YKZY001)the First Projects of Science and Technology Plan in the Biomedical field in 2021(202102AA310048)the National Natural Science Foundation of China(grant nos.81960691 and 82160727).
文摘Cucurbitacin IIa is a triterpenoid isolated exclusively from Hemsleya plants and a non-steroidal anti-inflammatory drug that functions as the main ingredient of prescription Hemslecin capsules and tablets in China.Synthetic biology provides new strategies for production of such valuable cucurbitacins at a large scale;however,the biosynthetic pathway of cucurbitacin IIa has been unknown,and the heterologous production of cucurbitacins in galactose medium has been expensive and low yielding.In this study,we characterized the functions of genes encoding two squalene epoxidases(HcSE1-2),six oxidosqualene cyclases(HcOSC1-6),two CYP450s(HcCYP87D20 and HcCYP81Q59),and an acyltransferase(HcAT1)in cucurbitacin IIa biosynthesis by heterologous expression in Saccharomyces cerevisiae and Nicotiana benthamiana.We achieved high-level production of the key cucurbitacin precursor 11-carbonyl-20b-hydroxy-Cuol from glucose in yeast via modular engineering of the mevalonate pathway and optimization of P450 expression levels.The resulting yields of 46.41 mg/l 11-carbonyl-20b-hydroxy-Cuol and 126.47 mg/l total cucurbitacin triterpenoids in shake flasks are the highest yields yet reported from engineered microbes.Subsequently,production of 11-carbonyl-20b-hydroxy-Cuol by transient gene expression in tobacco resulted in yields of 1.28 mg/g dry weight in leaves.This work reveals the key genes involved in biosynthesis of prescription cucurbitacin IIa and demonstrates that engineered yeast cultivated with glucose can produce high yields of key triterpenoid intermediates.We describe a low-cost and highly efficient platform for rapid screening of candidate genes and high-yield production of pharmacological triterpenoids.