检索结果-南通市图书馆

Engineering of β-carotene hydroxylase and ketolase for astaxanthin overproduction in Saccharomyces cerevisiae

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Frontiers of Chemical Science and Engineering 2017年第1期11卷 89-99页

作者： Ruizhao Wang Xiaoli Gu mingdong yao Caihui Pan Hong Liu Wenhai Xiao Ying Wang Yingjin Yuan Key Laboratory of Systems Bioengineering (Ministry of Education) Tianjin University Tianjin 300072 China SynBio Research Platform Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) School of Chemical Engineering and Technology Tianjin University Tianjin 300072 China

The conversion of r-carotene to astaxanthin is a complex pathway network, in which two steps of hydroxylation and two steps ofketolation are catalyzed by β-carotene hydroxylase （CrtZ） and β-carotene ketolase （CrtW） respectively. Here, astaxanthin biosynthesis path- way was constructed in Saccharomyces cerevisiae by introducing heterologous CrtZ and CrtW into an existing high r-carotene producing strain. Both genes crtZ and crtW were codon optimized and expressed under the control of constitutive promoters. Through combinatorial expression of CrtZ and CrtW from diverse species, nine strains in dark red were visually chosen from thirty combinations. In all the selected strains, strain SyBE Scl 18060 with CrtW from Brevundimonas vesicu- laris DC263 and CrtZ from Alcaligenes sp. strain PC-1 achieved the highest astaxanthin yield of 3.1 mg/g DCW. Protein phylogenetic analysis shows that the shorter evolutionary distance of CrtW is, the higher astaxanthin titer is. Further, when the promoter of crtZ in strain SyBE_Scl 18060 was replaced from FBAlp to TEFlp, the astaxanthin yield was increased by 30.4% （from 3.4 to 4.5 mg/g DCW）. In the meanwhile, 33.5-fold increase on crtZ transcription level and 39.1-fold enhancement on the transcriptional ratio of crtZ to crtWwere observed at early exponential phase in medium with 4% （w/v） glucose. Otherwise, although the ratio of crtZ to crtW were increased at mid-, late-exponential phases in medium with 2% （w/v） glucose, the transcription level of both crtZ and crtW were actually decreased during the whole timecourse, consequently leading to no significant improve- ment on astaxanthin production. Finally, through high cell density fed-batch fermentation using a carbon source restriction strategy, the production of astaxanthin in a 5-L bioreactor reached to 81.0 mg/L, which was the highest astaxanthin titer reported in yeast. This study provides a reference to greatly enhance lation by employing the key desired compounds accumu- enzyme

关键词： Keywords synthetic biology astaxanthin β-carotenehydroxylase,β-carotene ketolase Saccharomyces cerevi-siae

Enhanced Poly(ethylene terephthalate)Hydrolase Activity by Protein Engineering

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Engineering 2018年第6期4卷 888-893页

作者： Yuan Ma mingdong yao Bingzhi Li Mingzhu Ding Bo He Si Chen Xiao Zhou Yingjin Yuan Key Laboratory of Systems Bioengineering(Ministry of Education) School of Chemical Engineering and TechnologyTianjin UniversityTianjin 300072China SynBio Research Platform Collaborative Innovation Center of Chemical Science and EngineeringTianjin UniversityTianjin 300072China

Poly(ethylene terephthalate)hydrolase(PETase)from Ideonella sakaiensis exhibits a strong ability to degrade poly(ethylene terephthalate)(PET)at room temperature,and is thus regarded as a potential tool to solve the issue of polyester plastic ***,we explored the interaction between PETase and the substrate(a dimer of the PET monomer ethylene terephthalate,2PET),using a model of PETase and its *** this study,we focused on six key residues around the substrate-binding groove in order to create novel high-efficiency PETase mutants through protein *** PETase mutants were designed and *** enzymatic activities of the R61A,L88F,and I179F mutants,which were obtained with a rapid cell-free screening system,exhibited 1.4 fold,2.1 fold,and 2.5 fold increases,respectively,in comparison with wild-type *** I179F mutant showed the highest activity,with the degradation rate of a PET film reaching 22.5 mg perμmol·L^-1 PETase per ***,this study has created enhanced artificial PETase enzymes through the rational protein engineering of key hydrophobic sites,and has further illustrated the potential of biodegradable plastics.

关键词： Polyesterase PET degradation Cell-free protein synthesis Polyester PETase

Simulating androgen receptor selection in designer yeast

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Synthetic and Systems Biotechnology 2022年第4期7卷 1108-1116页

作者： Haoran Zhang Lu Zhang Yipeng Xu Shaoyong Chen Zhenyi Ma mingdong yao Fangyin Li Bo Li Yingjin Yuan Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering(Ministry of Education) Tianjin UniversityTianjin300072PR China School of Chemical Engineering and Technology Tianjin UniversityTianjin300072PR China Department of Urology The Cancer Hospital of the University of Chinese Academy of Sciences(Zhejiang Cancer Hospital)HangzhouChina Hematology-Oncology Division Department of MedicineBeth Israel Deaconess Medical Center and Harvard Medical SchoolBostonMA02215USA School of Basic Medical Sciences Tianjin Medical UniversityTianjin300070PR China

Androgen receptor(AR)mutation is closely associated with prostate cancer(PCa)and is one of the mechanisms of resistance to PCa therapies such as AR *** sequencing technologies like next-generation sequencing(NGS)contributes to the high-throughput and precise detection of AR mutations carried by PCa patients,the lack of interpretations of these clinical genetic variants has still been a roadblock for PCa-targeted precision ***,we established a designer yeast reporter assay to simulate natural androgen receptor(AR)selection using AR *** HIS3 gene transactivation was associated with the ligand-induced recruitment of steroid receptor coactivator-1(SRC-1)by AR mutants,where yeast growth in histidine-free medium was determined as the *** assay is applicable to determine a wide range of clinical AR mutants including those with loss of function relating to androgen insensitivity syndrome(AIS),and those associated with PCa conferring resistance to AR antagonists such as enzalutamide(ENZ),bicalutamide(BIC),and cyproterone acetate(CPA).One clinical AR mutant previously reported to confer ENZ-resistance,F877L,was found to confer partial resistance to CPA as well using designer *** simple and efficient assay can enable precise one-pot screening of AR mutants,providing a reference for tailored medicine.

关键词： Androgen receptor mutations Androgen receptor antagonists Prostate cancer Saccharomyces cerevisiae

Elucidating the Molecular Mechanism of Dynamic Photodamage of Photosystem II Membrane Protein Complex by Integrated Proteomics Strategy

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

CCS Chemistry 2022年第1期4卷 182-193页

作者： Ye Zhou Zheyi Liu mingdong yao Jun Chen Yanan Xiao Guangye Han Jian-Ren Shen Fangjun Wang CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023 State Key Laboratory of Catalysis Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023 Key Laboratory of Systems Bioengineering Ministry of EducationTianjin UniversityTianjin 300072 Science and Technology on Surface Physics and Chemistry Laboratory Jiangyou 621908 University of Chinese Academy of Sciences Beijing 100049 Photosynthesis Research Center Key Laboratory of PhotobiologyInstitute of BotanyChinese Academy of SciencesBeijing 100093 Research Institute for Interdisciplinary Science Graduate School of Natural Science and TechnologyOkayama UniversityOkayama 700-8530

Photosystem II(PSII),as a multiple-subunit chloroplast membrane-associated pigment-protein complex on the thylakoid membrane,is a primary target of light-induced ***,the overall molecular details of the conformation and composition dynamics of PSII photodamage are still *** this study,we investigated systematically the dynamic conformation,degradation,and oxidation processes of PSII photodamage by integrating chemical cross-linking and top-down proteomics strategies.

关键词： photosystem II dynamic photodamage chemical cross-linking top-down proteomics mass spectrometry

Engineering the Biosynthesis of Caffeic Acid in Saccharomyces cerevisiae with Heterologous Enzyme Combinations

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Engineering 2019年第2期5卷 287-295页

作者： Lanqing Liu Hong Liu Wei Zhang mingdong yao Bingzhi Li Duo Liu Yingjin Yuan Key Laboratory of Systems Bioengineering (Ministry of Education) School of Chemical Engineering and Technology Tianjin University Tianjin 300072 China SynBio Research Platform Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) Tianjin University Tianjin 300072 China

Engineering the biosynthesis of plant-derived natural products in microbes presents several challenges, especially when the expression and activation of the plant cytochrome P450 enzyme is required. By recruiting two enzymes—HpaB and HpaC—from several bacteria, we constructed functional 4- hydroxyphenylacetate 3-hydroxylase (4HPA3H) in Saccharomyces cerevisiae to take on a role similar to that of the plant-derived cytochrome P450 enzyme and produce caffeic acid. Along with a common tyrosine ammonia lyase (TAL), the different combinations of HpaB and HpaC presented varied capabilities in producing the target product, caffeic acid, from the substrate, L-tyrosine. The highest production of caffeic acid was obtained with the enzyme combination of HpaB from Pseudomonas aeruginosa and HpaC from Salmonella enterica, which yielded up to (289.4 ± 4.6) mg-L1 in shake-flask cultivation. The compatibility of heterologous enzymes within a yeast chassis was effectively improved, as the caffeic acid production was increased by 40 times from the initial yield. Six key amino acid residues around the flavin adenine dinucleotide (FAD) binding domain in HpaB from Pseudomonas aeruginosa were differentiate from those other HpaBs, and might play critical roles in affecting enzyme activity. We have thus established an effective approach to construct a highly efficient yeast system to synthesize non-native hydroxylated phenylpropanoids.

关键词： Saccharomyces cerevisiae Caffeic acid Heterologous enzyme Cytochrome P450 Synthetic biology

在线全文

学校读者我要写书评

暂无评论

生态多样性城市——印度尼西亚馆

建筑学报 2010年第6期 56-59页

作者：付睿姚鸣东上海市建工设计研究院有限公司上海200050

中国2010年上海世博会的主题是“城市，让生活更美好”，设计师将之阐释为“和谐的城市——即可持续发展的生态城市”。为了呼应本次世博会的主题，同时又体现印度尼西亚植物茂密、种类繁多，被誉为“赤道上的翡翠”的特点，印度尼西亚... 详细信息

中国2010年上海世博会的主题是“城市，让生活更美好”，设计师将之阐释为“和谐的城市——即可持续发展的生态城市”。为了呼应本次世博会的主题，同时又体现印度尼西亚植物茂密、种类繁多，被誉为“赤道上的翡翠”的特点，印度尼西亚馆根据展览建筑独特的坡道式形式，

关键词：印度尼西亚生态城市生态多样性 2010年上海世博会可持续发展展览建筑设计师坡道式

在线全文

学校读者我要写书评

暂无评论

维生素A生物合成的研究进展

化工学报 2022年第10期73卷 4311-4323页

作者：王欣慧王颖姚明东肖文海天津大学化工学院天津300072 天津大学合成生物学前沿科学中心和系统生物工程教育部重点实验室天津300072 天津大学前沿技术研究院天津301700

维生素A是维持人体代谢的必需维生素,在食品、药品及护肤品等多个领域占有重要地位,是三大维生素支柱产品之一,应用前景广阔。市场上的维生素A主要来源于化学合成和天然提取。近年来,随着绿色生物制造的发展,维生素A的生物合成研究取得... 详细信息

维生素A是维持人体代谢的必需维生素,在食品、药品及护肤品等多个领域占有重要地位,是三大维生素支柱产品之一,应用前景广阔。市场上的维生素A主要来源于化学合成和天然提取。近年来,随着绿色生物制造的发展,维生素A的生物合成研究取得了重大的进展。总结了维生素A生物合成的研究现状,并对维生素A生物合成的优化改造方案进行了分析归纳,概述了维生素A发酵生产以及产品组分调控和储存策略,最后对生物合成维生素A的现状进行总结与展望。

关键词：维生素A 细胞工厂合成生物学视黄醇酶代谢

同方期刊数据库博看期刊评论

在线全文

学校读者我要写书评

暂无评论

重组肌氨酸氧化酶的制备和鉴定

生物医学工程学杂志 2014年第5期31卷 1090-1096页

作者：蒲静王瑞姚明东何中杰赵明孟尧成都医学院第一附属医院成都610083 成都医学院检验医学院成都610500

测定血清中肌酐的浓度是临床上诊断肾功能的一项重要指标。在利用偶联酶法测定的过程中,一种关键酶——肌氨酸氧化酶(SOX)(EC 1.***.3.1)的质量控制是我们首先要解决的一个重要问题。本文利用乳糖诱导重组肌氨酸氧化酶(r-SOX)基因在***中... 详细信息

测定血清中肌酐的浓度是临床上诊断肾功能的一项重要指标。在利用偶联酶法测定的过程中,一种关键酶——肌氨酸氧化酶(SOX)(EC 1.***.3.1)的质量控制是我们首先要解决的一个重要问题。本文利用乳糖诱导重组肌氨酸氧化酶(r-SOX)基因在***中高效表达,经300L发酵罐发酵培养,菌体终密度达到OD600值22,菌体湿重达30g/L,湿菌体含活性20U/g。重组肌氨酸氧化酶表达量占菌体可溶性蛋白的25%左右。菌体破碎液经55℃选择性热变性处理和Ni-Sepharose FF层析二步纯化,经十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析酶的纯度达97%以上,比活力达25U/mg,纯化倍数为14.4,活性收率为92.4%。纯化的酶经SDS-PAGE和分子筛层析法测定,其分子量分别为53kD和55kD,提示该酶结构为单一肽链。纯化的酶液和冻干粉均呈黄色,经三氯乙酸和热变性处理,发现其与核黄素以共价键的方式结合。此外,对该酶的稳定性和其中的干扰酶过氧化氢酶也做了进一步的考察。以上所取得的研究结果为此酶的开发和利用奠定了基础。

关键词：重组肌氨酸氧化酶诱导表达高密度发酵

教育政策运行机制优化研究:超越理性与私利的视角

在线全文

学校读者我要写书评

暂无评论

现代教育管理 2017年第5期 69-74页

作者：黄明东姚宇华武汉大学湖北武汉430072

教育政策运行机制指教育政策的构成、功能、各组成部分的相互关系及其工作原理等多方面的内容,包括教育政策运行的所有阶段和环节的运行机制。考察理性和私利理论的发展历程,理性有工具理性和价值理性之分;私利理论的发展则证明人类不... 详细信息

教育政策运行机制指教育政策的构成、功能、各组成部分的相互关系及其工作原理等多方面的内容,包括教育政策运行的所有阶段和环节的运行机制。考察理性和私利理论的发展历程,理性有工具理性和价值理性之分;私利理论的发展则证明人类不仅有利己的一面,也有利他的另外一面。从理性和私利的角度看,我国教育政策运行机制存在理念上的"人性"缺失、模式上的科层性特征以及私利下的"公利性"偏离等困境。为了优化教育政策运行机制,在超越理性和私利的指导下,应不断平衡工具理性和价值理性,加强公共部门及其成员私利追求的规制和完善社会参与机制。

关键词：教育政策运行机制优化超越理性和私利工具理性价值理性