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Functional characterization of CYP81C16 involved in the tanshinone biosynthetic pathway in Salvia miltiorrhiza

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Chinese Journal of Natural Medicines 2023年第12期21卷 938-949页

作者： REN Li LUO Linglong HU Zhimin MA Ying WANG Jian CHENG Yatian jin baolong CHEN Tong TANG jinfu CUI Guanghong GUO Juan HUANG Luqi State Key Laboratory of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100000China

Danshen,the dried roots and rhizomes of Salvia miltiorrhiza Bunge(***),is widely used in the treatment of cardiovascular and cerebrovascular ***,the bioactive compounds from Danshen,exhibit a wide spectrum of pharmacological properties,suggesting their potential for future therapeutic *** 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 *** this study,CYP81C16,a member of the CYP71 clan,was identified in *** in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones,namely neocryptotanshinone,deoxyneocryptotanshinone,and danshenxinkuns A and ***81C16 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 *** overexpression of SmCYP81C16 promoted the accumulation of(iso)tanshinone in hairy root *** 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.

关键词： Salvia miltiorrhiza Tanshinone Biosynthetic pathway Cytochrome P450 Hydroxylation Neocryptotanshinone

Functional identification of the terpene synthase family involved in diterpenoid alkaloids biosynthesis in Aconitum carmichaelii

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Acta Pharmaceutica Sinica B 2021年第10期11卷 3310-3321页

作者： 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 College of Pharmacy Shandong University of Chinese MedicineJinan 250355China State Key Laboratory of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China

Aconitum carmichaelii is a high-value medicinal herb widely used across China,Japan,and other Asian ***-type diterpene alkaloids(DAs)are the characteristic compounds in *** 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 *** apply a combination of Pacbio isoform sequencing and RNA sequencing to provide a comprehensive view of the *** *** TPSs and five alternative splicing isoforms belonging to TPS-b,TPS-c,and TPS-e/f subfamilies were *** vitro enzyme reaction analysis functional identified two sesqui-TPSs and twelve di *** of the TPS-c subfamily genes reacted with GGPP to produce the intermediate ent-copalyl *** Ac KSLs separately reacted with ent-CPP to produce ent-kaurene,ent-atiserene,and ent-13-epi-sandaracopimaradie:a new diterpene found in *** 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,*** data clarified the molecular basis for the C20-DAs biosynthetic pathway in *** and pave the way for further exploration of C19-DAs biosynthesis in the Aconitum species.

关键词： Aconitum carmichaelii Full-length transcriptome Diterpene synthase Diterpenoid alkaloids

Identification of (-)-bornyl diphosphate synthase from Blumea balsamifera and its application for (-)-borneol biosynthesis in Saccharomyces cerevisiae

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Synthetic and Systems Biotechnology 2022年第1期7卷 490-497页

作者： Rui Ma Ping Su Qing Ma Juan Guo Suiqing Chen baolong jin Haiyan Zhang jinfu Tang Tao Zhou Chenghong Xiao Guanghong Cui Luqi Huang School of Pharmacy Henan University of Chinese MedicineZhengzhouChina State Key Laboratory Breeding Base of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijingChina School of Pharmacy Guizhou University of Traditional Chinese MedicineGuiyangChina

Borneol is a precious monoterpenoid with two chiral structures,(-)-borneol and(+)-*** diphosphate synthase is the key enzyme in the borneol biosynthesis ***(+)-bornyl diphosphate synthases have been reported,but no(-)-bornyl diphosphate synthases have been *** balsamifera leaves are rich in borneol,almost all of which is(-)-*** this study,we identified a high-efficiency(-)-bornyl diphosphate synthase(BbTPS3)from *** that converts geranyl diphosphate(GPP)to(-)-bornyl diphosphate,which is then converted to(-)-borneol after dephosphorylation in ***3 exhibited a K m value of 4.93±1.38μM for GPP,and the corresponding k cat value was 1.49 s−*** 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)(-)-*** 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 *** work is the first reported attempt to produce(-)-borneol by microbial fermentation.

关键词： Metabolic engineering (-)-borneol (-)-bornyl diphosphate synthase Blumea balsamifera Saccharomyces cerevisiae

永磁同步电机模糊遗忘因子最小二乘法参数辨识

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系统仿真学报 2018年第9期30卷 3404-3410,3419页

作者：沈艳霞靳保龙江南大学物联网工程学院电气自动化研究所江苏无锡214122

电机节能控制中损耗模型法要求电机参数尽可能稳定准确,为提高永磁同步电机参数在线辨识的稳定性及收敛速度,基于带遗忘因子的最小二乘算法,提出一种模糊遗忘因子最小二乘算法。首先利用帕德逼近法线性化技术建立永磁同步电机线性回归... 详细信息

电机节能控制中损耗模型法要求电机参数尽可能稳定准确,为提高永磁同步电机参数在线辨识的稳定性及收敛速度,基于带遗忘因子的最小二乘算法,提出一种模糊遗忘因子最小二乘算法。首先利用帕德逼近法线性化技术建立永磁同步电机线性回归数学模型,根据电流误差设计模糊控制器,进行遗忘因子的自适应调整,并将其应用于永磁同步电机定子电阻的在线辨识中,较好地解决了遗忘因子最小二乘算法中结果稳定性和收敛速度相互矛盾的问题,最后仿真验证了该方法的有效性。

关键词：遗忘因子最小二乘模糊控制参数辨识永磁同步电机

Identification of the cytochrome P450s responsible for the biosynthesis of two types of aporphine alkaloids and their de novo biosynthesis in yeast

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Journal of Integrative Plant Biology 2024年第8期66卷 1703-1717页

作者： Qishuang Li Xiang Jiao Xinyi Li Wenlong Shi Ying Ma Xiangmei Tan jingyi Gan Jimei Liu Jian Yang Jian Wang baolong jin Tong Chen Ping Su Yujun Zhao Yifeng Zhang jinfu Tang Guanghong Cui Yun Chen Juan Guo Luqi Huang School of Traditional Chinese Pharmacy China Pharmaceutical UniversityNanjing 211198China State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China Department of Life Sciences Chalmers University of TechnologyGothenburg SE-41296Sweden State Key Laboratory of Natural and Biomimetic Drugs School of Pharmaceutical SciencesPeking UniversityBeijing 100191China State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaThe Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China

Aporphine alkaloids have diverse pharmacological activities;however,our understanding of their biosynthesis is relatively *** studies have classified aporphine alkaloids into two categories based on the configuration and number of substituents of the D-ring and have proposed preliminary biosynthetic pathways for each *** this study,we identified two specific cytochrome P450 enzymes(CYP80G6 and CYP80Q5)with distinct activities toward(S)-configured and(R)-configured substrates from the herbaceous perennial vine Stephania tetrandra,shedding light on the biosynthetic mechanisms and stereochemical features of these two aporphine alkaloid ***,we characterized two CYP719C enzymes(CYP719C3 and CYP719C4)that catalyzed the formation of the methylenedioxy bridge,an essential pharmacophoric group,on the A-and D-rings,respectively,of aporphine *** the functional characterization of these crucial cytochrome P450 enzymes,we reconstructed the biosynthetic pathways for the two types of aporphine alkaloids in budding yeast(Saccharomyces cerevisiae)for the de novo production of compounds such as(R)-glaziovine,(S)-glaziovine,and *** study provides key insight into the biosynthesis of aporphine alkaloids and lays a foundation for producing these valuable compounds through synthetic biology.

关键词： aporphine alkaloids biosynthesis CYP719C CYP80 engineered yeast

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多机协同作业全覆盖路径规划

农机化研究 2024年第12期46卷 28-33页

作者：金宝龙夏长高韩江义江苏大学汽车与交通工程学院江苏镇江2120133

多机协同作业能够提升作业效率,节约作业时间,减少了农作物因收获不及时导致的粮食浪费。为此,以总的非工作距离和最长单车行驶距离为目标函数,建立虚拟的农田模型,将多机协同作业全覆盖路径规划问题转化为VRP问题,并使用改进的蚁群算... 详细信息

多机协同作业能够提升作业效率,节约作业时间,减少了农作物因收获不及时导致的粮食浪费。为此,以总的非工作距离和最长单车行驶距离为目标函数,建立虚拟的农田模型,将多机协同作业全覆盖路径规划问题转化为VRP问题,并使用改进的蚁群算法求解。仿真结果表明:改进后的算法有效;对比遗传算法和传统的蚁群算法,改进后的算法平均目标函数值分别降低了25.0%和11.25%;对比模拟退火算法,改进后的算法平均目标函数值降低了1.5%,算法稳定性更好,适用于更大规模农田的多机协同作业全覆盖路径规划问题求解。

关键词：多机协同全覆盖路径规划蚁群算法优化算法

Functional characterization of seven terpene synthases from Ophiopogon japonicus via engineered Escherichia coli

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Science of Traditional Chinese Medicine 2024年第4期2卷 312-318页

作者： Wen Xu Qing Ma Tong Chen Haiyan Zhang Guanglei Ma Sibongile Mafu Juan Guo Xiaohui Fan Guanghong Cui baolong jin State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijingChina College of Pharmaceutical Sciences Zhejiang UniversityHangzhouChina National Key Laboratory of Chinese Medicine Modernization Innovation Center of Yangtze River DeltaZhejiang UniversityJiaxingChina Department of Biochemistry and Molecular Biology University of Massachusetts-AmherstAmherstMAUnited States

Background:Ophiopogon japonicus(L.f)*** in Zhejiang is recognized as the Dao-di medicinal herb for the production of Ophiopogonis ***-7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside,a prominent pharmacologically active compound,serves as a marker distinguishing *** in Zhejiang from those in other geographical *** is synthesized from borneol through glycosylation,with terpene synthase(TPS)being the critical enzyme catalyzing the conversion of terpene precursors into ***:The aim of the study was to define key genes involved in biosynthesis of borneol in ***:The candidate terpene synthase genes were identified from the root and leaf transcriptome data of *** in Zhejiang and the functions of these enzymes were identified using engineered Escherichia ***:This study developed a rapid expression system for monoterpene and sesquiterpene synthases based on engineered *** terpene synthase genes(OjTPS1 to OjTPS7)were identified in different terpene synthase subfamilies,including 2 from TPS-a,4 from TPS-b,and 1 from *** analysis using an engineered system *** demonstrated that all the 7 terpene synthases produced monoterpenes,and OjTPS3,OjTPS5,and OjTPS6 also yielded ***:These 7 terpene synthases produced 13 monoterpenes and 8 ***,OjTPS1 produced borneol establishing the groundwork for elucidating the biosynthetic pathways of borneol-7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside and other volatile oil components.

关键词： Ophiopogon japonicus Ophiopogon japonicus in Zhejiang Terpene synthase Borneol

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一种新的小波阈值去噪算法

弹箭与制导学报 2011年第1期31卷 167-169,176页

作者：金宝龙李辉赵乃杰何海峰西北工业大学电子信息学院西安710129 骊山微电子公司西安710065

雷达一维高分辨距离像对噪声极为敏感。研究一种新的小波去噪算法得到高信噪比的雷达一维高分辨距离像是实现精确目标识别的关键。文中对比分析了小波模极大值去噪算法和小波阈值去噪算法,并在经典小波阈值去噪算法的基础上改进了阈值函... 详细信息

雷达一维高分辨距离像对噪声极为敏感。研究一种新的小波去噪算法得到高信噪比的雷达一维高分辨距离像是实现精确目标识别的关键。文中对比分析了小波模极大值去噪算法和小波阈值去噪算法,并在经典小波阈值去噪算法的基础上改进了阈值函数,提出了一种新小波阈值去噪算法。实验证明新的算法能够更好的提高雷达一维高分辨距离像的信噪比,有效抑制噪声敏感性。新算法结合了软、硬阈值算法的优点,灵活度高。

关键词：小波去噪阈值模极大值雷达一维高分辨距离像

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混凝土腐蚀破坏的反应机理分析

江西建材 2018年第10期 22-23,8页

作者：靳宝龙范建军东方雨虹民用建材有限责任公司北京100176

海洋及盐碱地等恶劣环境对钢筋混凝土建筑物或构筑物的腐蚀问题日益严重,防止钢筋混凝土结构的腐蚀和增强其耐久性越加重要。本文对混凝土腐蚀破坏机理进行分析,对混凝土结构采取有效的防护技术,对保证并提高混凝土结构的耐久性与使用... 详细信息

海洋及盐碱地等恶劣环境对钢筋混凝土建筑物或构筑物的腐蚀问题日益严重,防止钢筋混凝土结构的腐蚀和增强其耐久性越加重要。本文对混凝土腐蚀破坏机理进行分析,对混凝土结构采取有效的防护技术,对保证并提高混凝土结构的耐久性与使用寿命具有重要的现实意义。

关键词：防腐阻锈硫酸盐侵蚀反应机理