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| 1 | Functional identification of the terpene synthase family involved in diterpenoid alkaloids biosynthesis in Aconitum carmichaelii显示文摘Aconitum carmichaelii is a high-value medicinal herb widely used across China,Japan,and other Asian countries.Aconitine-type diterpene alkaloids(DAs)are the characteristic compounds in Aconitum.Although six transcriptomes,based on short-read next generation sequencing technology,have been reported from the Aconitum species,the terpene synthase(TPS)corresponding to DAs biosynthesis remains unidentified.We apply a combination of Pacbio isoform sequencing and RNA sequencing to provide a comprehensive view of the A.carmichaelii transcriptome.Nineteen TPSs and five alternative splicing isoforms belonging to TPS-b,TPS-c,and TPS-e/f subfamilies were identified.In vitro enzyme reaction analysis functional identified two sesqui-TPSs and twelve di TPSs.Seven of the TPS-c subfamily genes reacted with GGPP to produce the intermediate ent-copalyl diphosphate.Five Ac KSLs separately reacted with ent-CPP to produce ent-kaurene,ent-atiserene,and ent-13-epi-sandaracopimaradie:a new diterpene found in Aconitum.Ac TPSs gene expression in conjunction DAs content analysis in different tissues validated that ent-CPP is the sole precursor to all DAs biosynthesis,with Ac KSL1,Ac KSL2 s and Ac KSL3-1 responsible for C20 atisine and napelline type DAs biosynthesis,respectively.These data clarified the molecular basis for the C20-DAs biosynthetic pathway in A.carmichaelii and pave the way for further exploration of C19-DAs biosynthesis in the Aconitum species. | Liuying Mao Baolong Jin Lingli Chen Mei Tian Rui Ma Biwei Yin Haiyan Zhang Juan Guo Jinfu Tang Tong Chen Changjiangsheng Lai Guanghong Cui Luqi Huang | 2021 | Acta Pharmaceutica Sinica B2021，11，10： | 6 |
| 2 | The rapid uncertainty prediction of the ocean-acoustic coupled model显示文摘 | CUI Baolong DA Lianglong GUO Wuhong ZHAO Jianxin | 2018 | Chinese Journal of Acoustics2018，37，4： | 2 |
| 3 | Calcium ions promote primary renal epithelial cell differentiation intocells with bone-associated phenotypes via transforming growth factor-β1-inducedepithelial-mesenchymal transition in idiopathic hypercalciuria patients显示文摘 | Deng He Shaogang Wang Zhaohui Jia Lei Cui Yuchao Lu Henglong Hu Baolong Qin | 2015 | Molecular Medicine Reports2015，，3： | 1 |
| 4 | Identification of the sensitive area for targeted observation to improve vertical thermal structure prediction in summer in the Yellow Sea显示文摘The sensitive area of targeted observations for short-term(7 d)prediction of vertical thermal structure(VTS)in summer in the Yellow Sea was investigated.We applied the Conditional Nonlinear Optimal Perturbation(CNOP)method and an adjoint-free algorithm with the Regional Ocean Modeling System(ROMS).We used vertical integration of CNOP-type temperature errors to locate the sensitive areas,where reduction of initial errors is expected to yield the greatest improvement in VTS prediction for the selected verification area.The identified sensitive areas were northeast−southwest orientated northeast to the verification area,which were possibly related to the southwestward background currents.Then,we performed a series of sensitivity experiments to evaluate the effectiveness of the identified sensitive areas.Results show that initial errors in the identified sensitive areas had the greatest negative effect on VTS prediction in the verification area compared to errors in other areas(e.g.,the verification area and areas to its east and northeast).Moreover,removal of initial errors through deploying simulated observations in the identified sensitive areas led to more refined prediction than correction of initial conditions in the verification area itself.Our results suggest that implementation of targeted observation in the CNOP-based sensitive areas is an effective method to improve short-term prediction of VTS in summer in the Yellow Sea. | Huiqin Hu Jingyi Liu Lianglong Da Wuhong Guo Kun Liu Baolong Cui | 2021 | Acta Oceanologica Sinica2021，40，7： | 1 |
| 5 | Identification of (-)-bornyl diphosphate synthase from Blumea balsamifera and its application for (-)-borneol biosynthesis in Saccharomyces cerevisiae显示文摘Borneol is a precious monoterpenoid with two chiral structures,(-)-borneol and(+)-borneol.Bornyl diphosphate synthase is the key enzyme in the borneol biosynthesis pathway.Many(+)-bornyl diphosphate synthases have been reported,but no(-)-bornyl diphosphate synthases have been identified.Blumea balsamifera leaves are rich in borneol,almost all of which is(-)-borneol.In this study,we identified a high-efficiency(-)-bornyl diphosphate synthase(BbTPS3)from B.balsamifera that converts geranyl diphosphate(GPP)to(-)-bornyl diphosphate,which is then converted to(-)-borneol after dephosphorylation in vitro.BbTPS3 exhibited a K m value of 4.93±1.38μM for GPP,and the corresponding k cat value was 1.49 s−1.Multiple strategies were applied to obtain a high-yielding(-)-borneol producing yeast strain.A codon-optimized BbTPS3 protein was introduced into the GPP high-yield strain MD,and the resulting MD-B1 strain produced 1.24 mg⋅L^(-1)(-)-borneol.After truncating the N-terminus of BbTPS3 and adding a Kozak sequence,the(-)-borneol yield was further improved by 4-fold to 4.87 mg⋅L^(-1).Moreover,the(-)-borneol yield was improved by expressing the fusion protein module of ERG20 F96W-N127W-YRSQI-t14-BbTPS3K2,resulting in a final yield of 12.68 mg⋅L^(-1) in shake flasks and 148.59 mg⋅L^(-1) in a 5-L bioreactor.This work is the first reported attempt to produce(-)-borneol by microbial fermentation. | Rui Ma Ping Su Qing Ma Juan Guo Suiqing Chen Baolong Jin Haiyan Zhang Jinfu Tang Tao Zhou Chenghong Xiao Guanghong Cui Luqi Huang | 2022 | Synthetic and Systems Biotechnology2022，7，1： | 0 |
| 6 | Functional characterization of CYP81C16 involved in the tanshinone biosynthetic pathway in Salvia miltiorrhiza显示文摘Danshen,the dried roots and rhizomes of Salvia miltiorrhiza Bunge(S.miltiorrhiza),is widely used in the treatment of cardiovascular and cerebrovascular diseases.Tanshinones,the bioactive compounds from Danshen,exhibit a wide spectrum of pharmacological properties,suggesting their potential for future therapeutic applications.Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized,most of which belong to the CYP76 and CYP71 families.In this study,CYP81C16,a member of the CYP71 clan,was identified in S.miltiorrhiza.An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones,namely neocryptotanshinone,deoxyneocryptotanshinone,and danshenxinkuns A and B.SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%.Kinetic evaluations and in vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates.The overexpression of SmCYP81C16 promoted the accumulation of(iso)tanshinone in hairy root lines.The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones,either through microbial or plant metabolic engineering. | REN Li LUO Linglong HU Zhimin MA Ying WANG Jian CHENG Yatian JIN Baolong CHEN Tong TANG Jinfu CUI Guanghong GUO Juan HUANG Luqi | 2023 | Chinese Journal of Natural Medicines2023，21，12： | 0 |
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