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Establishing Modular Cell-Free Expression System for the Biosynthesis of Bicyclomycin from a Chemically Synthesized Cyclodipeptide

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Chinese Journal of Chemistry 2024年第4期42卷 384-390页

作者： Yi-Pei Liu Yu-Heng Zhao Wen-Qi Zhang Lian Wu linjuan huang Gong-Li Tang Jun-Bin He Shaanxi Natural Carbohydrate Resource Engineering Research Center College of Food Science and TechnologyNorthwest UniversityXi'anShaanxi710069 China State Key Laboratory of Chemical Biology Shanghai Institute of Organic ChemistryUniversity of Chinese Academy of Sciences(CAS)CASShanghai200032 China School of Chemistry and Materials Science Hangzhou Institute for Advanced StudyUniversity of CAS1 Sub-lane XiangshanHangzhouZhejiang310024 China

Cell-free expression systems have emerged as a versatile and powerful platform for metabolic engineering,biosynthesis and synthetic biology ***,successful examples of the synthesis of complex natural products using this system are still ***,a structurally unique and complex diketopiperazine alkaloid,is a clinically promising antibiotic that selectively inhibits the transcription termination factor ***,we established a modular cell-free expression system with cascade catalysis for the biosynthesis of bicyclomycin from a chemically synthesized *** six cell-free expressed biosynthetic enzymes,including five iron-andα-ketoglutarate-dependent dioxygenases and one cytochrome P450 monooxygenase,were active in converting their substrates to the corresponding *** co-expressed enzymes in the cell-free module were able to complete the related partial *** vitro biosynthesis of bicyclomycin was also achieved by reconstituting the entire biosynthetic pathways(i.e.,six enzymes)using the modular cell-free expression *** study demonstrates that the modular cell-free expression system can be used as a robust and promising platformforthe biosynthesis of complex antibiotics.

关键词： Natural products Bicyclomycin Biosynthesis Iron-andα-ketoglutarate-dependent dioxygenases Cytochrome P450 monooxygenase Cell-freeprotein synthesis Enzyme catalysis

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An innovative method used for the identification of N-glycans on soybean allergenβ-conglycinins

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Food Science and Human Wellness 2023年第3期12卷 842-850页

作者： Cheng Li Yang Tian Jianli Han Yu Lu Meiyi Zou Yue Jia Chengjian Wang linjuan huang Zhongfu Wang Key Laboratory of Glycobiology and Glycoengineering of Xi’an College of Food Science and TechnologyNorthwest UniversityXi’an 710069China College of Life Sciences Northwest UniversityXi’an 710069China

β-Conglycinin is one of the major allergens existed in soybean.N-Glycans attached to theβ-conglycinin influenced the immunoreactivity and antigen presenting efficiency ofβ-*** this study,we described a new method used to release and collect the N-glycans fromβ-conglycinin,and the N-glycans existed in linear epitopes ofβ-conglycinin were *** hydrolyzed fromβ-conglycinin were purified by cotton hydrophilic *** acid was then used to release glycans from glycopeptides,and new glycopeptides containing one single N-acety1-D-glucosamine(G1 cNAc)moiety were then utilized for mass *** glycosylation sites(Asn-199,Asn-455,Asn-215,Asn-489 and Asn-326)and 22 kinds of glycopeptides were *** is noteworthy that the peptide VVN^(#)ATSNL(where^(#)represents for the glycosylation site)was analyzed to be both glycopeptide and linear *** results provided a new method for the N-glycoform analysis of food allergens,and laid a foundation for understanding the relationship between glyco sylation and food allergy.

关键词： Soybean allergenβ-conglycinin Glycopeptide Mass spectrometry N-Glycan Glycosylation site

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Progress in The Treatment of Silicosis（Review）

Progress in The Treatment of Silicosis（Review）

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第七届全国呼吸系统重大疾病转化医学学术论坛

作者： huang linjuan Xu Jianping Luo ziqiang Central South University School of Basia Medicine

Silicosis is one of the most serious occupational diseases in the *** lavage and cell transplantation are expected to be effective ways to cure silicosis,repair pulmonary fibrosis and reconstruct lung *** article revi... 详细信息

关键词： Silicosis Treatment Irrigation Stem cell therapy

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Chemical synthesis of UDP-2-acetonyl-Gal from its corresponding natural saccharide

Chemical synthesis of UDP-2-acetonyl-Gal from its correspond...

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第二届糖合成化学青年学术研讨会

作者：张英 Mingming Yang Hongdan Shi linjuan huang Zhongfu Wang 西北大学 Key Laboratory of Resource Biology and Biotechnology in Western China Ministry of Education College of Life Sciences Northwest University

O-GlcNAcylations,one of the most universal and important post-translational modifications of protein,take charge of plenty of cellular-biological processes and are associated with variety of diseases such as diabetes and tumor and Alzheimer’s disease when their glycosylations are at abnormal expression ***,the detection of intracellular protein O-Glc NAc glycosylated modifications,can be used as a research reference for the study of occurrence and development of disease and pathogenesis[1].Due to the structure complexity and micro-scale expression within the cell,the detecting and analyzing O-Glc NAc glycosylation have suffered from some drawbacks including lack of selective assay for detection or capturing the low-abundance O-Glc NAcylated ***,the comprehensive studies on function-structure mechanism of protein O-Glc NAcylation that involve in disease development progress have been seriously hindered these years.A chemical-enzymic reaction that could enable to add active acetonyl-containing Galactose（UDP-2-keto-Gal）to C-4 position of O-Glc NAc moiety on OGlc NAcylation protein,provide an effective avenue for selectively and efficiently detecting the target glycoprotein by using hydrazide containing chemicals[2].However,the ketone-substrate analogues are not commercialized,leading to that the detection of the O-Glc NAcylated modification is severely *** present study focused on the establishment of a universal chemical method for the synthesis of ketone substrate *** our work,we mainly made these innovations as follows:（1）The synthesis of glycal from Gal was conducted by an one-pot method without the need of purification,which could reduce the loss of sample and simplify the reaction steps;（2）Osmium tetroxide and sodium periodate method is used to replace ozone oxidation of carbon-carbon double bond,to generate keto sugar under room temperature;（3）In the synthesis of UDP-ketone-sugar,the C-1 was converted to be the trichloroac

关键词： O-GlcNAc UDP-2-acetonyl-Gal chemical synthesis

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Niclosamide(NA)overcomes cisplatin resistance in human ovarian cancer

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Genes & Diseases 2023年第4期10卷 1687-1701页

作者： linjuan huang Jing Zhang Youling Deng Hao Wang Piao Zhao Guozhi Zhao Wei Zeng Yonghui Wang Connie Chen William Wagstaff Rex C.Haydon Russell R.Reid Tong-Chuan He Le Shen Hue H.Luu Ling Zhao Departments of Obstetrics and Gynecology Orthopaedic Surgery and UrologyThe First Affiliated Hospital of Chongqing Medical UniversityChongqing 400046China Molecular Oncology Laboratory Department of Orthopaedic Surgery and Rehabilitation MedicineThe University of Chicago Medical CenterChicagoIL 60637USA Ministry of Education Key Laboratory of Diagnostic Medicine and Department of Clinical BiochemistryThe School of Laboratory MedicineChongqing Medical UniversityChongqing 400016China Department of Neurology The Second Affiliated Hospital of Jianghan UniversityWuhanHubei 430050China Department of Clinical Laboratory Medicine Shanghai Jiaotong University School of MedicineShanghai 200000China Laboratory of Craniofacial Suture Biology and Development Department of Surgery Section of Plastic SurgeryThe University of Chicago Medical CenterChicagoIL 60637USA Department of Surgery The University of Chicago Medical CenterChicagoIL 60637USA

Ovarian cancer(OC)is one of the most lethal malignancies of the female reproduc-tive *** patients are usually diagnosed at advanced stages due to the lack of early *** standard treatment for OC includes a combination of debulking surgery and platinum-taxane chemotherapy,while several targeted therapies have recently been approved for maintenance *** vast majority of OC patients relapse with chemoresistant tu-mors after an initial ***,there is an unmet clinical need to develop new therapeu-tic agents to overcome the chemoresistance of *** anti-parasite agent niclosamide(NA)has been repurposed as an anti-cancer agent and exerts potent anti-cancer activities in human cancers including ***,we investigated whether NA could be repurposed as a therapeutic agent to overcome cisplatin-resistant(CR)in human OC *** this end,we first established two CR lines SKOV3CR and OVCAR8CR that exhibit the essential biological characteristics of cisplatin resistance in human *** showed that NA inhibited cell proliferation,sup-pressed cell migration,and induced cell apoptosis in both CR lines at a low micromole ***,NA inhibited multiple cancer-related pathways including AP1,ELK/SRF,HIF1,and TCF/LEF,in SKOV3CR and OVCAR8CR *** was further shown to effectively inhibit xenograft tumor growth of SKOV3CR ***,our findings strongly suggest that NA may be repurposed as an efficacious agent to combat cisplatin resistance in chemoresistant hu-man OC,and further clinical trials are highly warranted.

关键词： Chemotherapy resistance Cisplatin Drug repurposing Niclosamide Ovarian cancer

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糖蛋白的凝胶电泳和电印迹染色鉴定技术

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化学进展 2008年第7期20卷 1158-1164页

作者：张英黄琳娟王仲孚西北大学生命科学学院西部资源生物与现代生物技术省部共建教育部重点实验室陕西省生物技术重点实验室西安710069

糖基化是蛋白质翻译后修饰最重要的方式之一,是必需的生理过程,对蛋白质生物学功能的正确发挥起着至关重要的作用。二维凝胶电泳(two-dimensional gel electrophoresis,2-DE)是常用的分离蛋白质的方法之一,糖蛋白在凝胶上的检测是分离... 详细信息

糖基化是蛋白质翻译后修饰最重要的方式之一,是必需的生理过程,对蛋白质生物学功能的正确发挥起着至关重要的作用。二维凝胶电泳(two-dimensional gel electrophoresis,2-DE)是常用的分离蛋白质的方法之一,糖蛋白在凝胶上的检测是分离鉴定糖蛋白的关键之一。本文对近年来凝胶电泳、电印迹技术中鉴定糖蛋白的衍生试剂的研究和应用进行了综述,对多种衍生试剂衍生糖蛋白的机理作了介绍。

关键词：糖蛋白凝胶电泳电印迹染色

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Highly Sensitive MALDI-TOF-MS Analysis of Reducing Glycan by On-target Derivatization with Hydrazines

Highly Sensitive MALDI-TOF-MS Analysis of Reducing Glycan by...

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中国生物工程学会第十一届学术年会暨2017年全国生物技术大会

作者： Ying Zhang Bo Wang Wanjun Jin Chengjian Wang linjuan huang Zhongfu Wang Key Laboratory of Resource Biology and Biotechnology in Western China Ministry of Education and Provincial Key Laboratory of Biotechnology College of Life Sciences Northwest University

Glycan have been found to participate in many important biological *** analysis of glycans by matrixassisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) is of great importance but significantly hampered by their low ionization efficiency or sialic acid *** derivatization offer an viable way to improve both the ionization efficiency and analytical sensitivity of the sugar sample in MS analysis by altering theirhydrophobicity or charge ability.

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荧光辅助糖电泳用于果胶水解产物中寡糖的分析研究

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化学学报 2009年第9期67卷 951-956页

作者：孙静李健军黄琳娟王仲孚西北大学生命科学学院西部资源生物与现代生物技术省部共建教育部重点实验室西北大学糖生物学与糖工程研究中心西安710069 西安交通大学理学院化学系西安710049

荧光辅助糖电泳(FACE)是首先用荧光衍生化试剂对糖类分子的还原端进行衍生化标记,然后在一定浓度的聚丙烯酰胺凝胶上进行分离的分析方法,可同时分析中性糖与酸性糖.将该方法用于果胶寡糖的分离分析,对影响FACE的诸多因素如荧光试剂的种... 详细信息

荧光辅助糖电泳(FACE)是首先用荧光衍生化试剂对糖类分子的还原端进行衍生化标记,然后在一定浓度的聚丙烯酰胺凝胶上进行分离的分析方法,可同时分析中性糖与酸性糖.将该方法用于果胶寡糖的分离分析,对影响FACE的诸多因素如荧光试剂的种类、用量,荧光衍生化时间、温度,分离胶浓度及盐、酸等进行了考察,对果胶寡糖的FACE条件进行了优化,结果显示:每1.2mg无酸且不含盐的果胶寡糖中,加入0.2mol/L的ANTS溶液3.75μL,1.0mol/L的NaBH3CN溶液5μL,40℃衍生化反应16h后,在浓度为38%的分离胶上电泳分离,取得良好的分离效果.在该实验条件下,果胶多糖酸水解后得到聚合度为2～16的果胶寡糖混合物,与质谱分析结果基本一致.该方法快速、简捷、灵敏、分辨率高,费用低,为果胶多糖可控性降解的监测和果胶寡糖的分离分析提供了技术手段.

关键词：果胶寡糖衍生物荧光辅助糖电泳 8-氨基萘-1, 3 ,6-三磺酸 2-氨基吖啶酮

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学龄前营养不良儿童血清25-(OH)D_(3)、白蛋白、铁蛋白与STAMP评分的相关性及对临床结局的预测价值

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中国儿童保健杂志 2024年第1期32卷 39-44页

作者：宋捷黄林娟郭碧丹福建省福州儿童医院临床营养科福建福州350005 福建省福州儿童医院儿保科福建福州350005

目的探讨学龄前营养不良儿童血清25-(OH)D_(3)、白蛋白(ALB)、铁蛋白(SF)与儿科营养不良评估筛查工具(STAMP)评分的相关性,并分析对临床结局的预测价值,为改善营养不良的预后提供参考。方法选取2021年8月—2022年8月福州儿童医院90例学... 详细信息

目的探讨学龄前营养不良儿童血清25-(OH)D_(3)、白蛋白(ALB)、铁蛋白(SF)与儿科营养不良评估筛查工具(STAMP)评分的相关性,并分析对临床结局的预测价值,为改善营养不良的预后提供参考。方法选取2021年8月—2022年8月福州儿童医院90例学龄前营养不良患儿作为观察组,另选取同期学龄前健康体检儿童60例作为对照组。学龄前儿童予以营养指导,连续3个月,随访统计治疗后3个月内营养不良及缺铁性贫血发生情况。使用STAMP对所有研究对象进行评估,分析血清25-(OH)D_(3),ALB、SF与学龄前营养不良患儿STAMP评分的关联性,并评价三者联合检测对营养不良临床结局的预测价值。结果观察组血清25-(OH)D_(3)、ALB、SF水平均低于对照组,STAMP评分高于对照组(t=9.700、6.047、4.771、26.976,P<0.05);学龄前营养不良患儿血清25-(OH)D_(3)、ALB、SF水平与STAMP评分均呈负相关(r=-0.813、-0.792、-0.829,P<0.05);治疗3个月后,临床结局不良患儿血清25-(OH)D_(3)、ALB、SF及其治疗前后差值均低于临床结局良好患儿(t=3.622、2.189、2.163、8.704、4.354、5.296,P<0.05);学龄前营养不良患儿血清25-(OH)D_(3)、ALB、SF治疗前后差值处于低水平时,其临床结局不良风险分别是高水平的7.250倍(95%CI:3.008~17.476)、9.167倍(95%CI:2.954~28.444)、5.730倍(95%CI:2.364~13.890);血清25-(OH)D_(3)、ALB、SF治疗前后差值联合预测的AUC值高于单一指标,为0.927;DCA曲线结果显示血清25-(OH)D_(3)、ALB、SF治疗前后差值联合预测方案预测学龄前营养不良患儿临床结局具有临床净获益。结论低水平血清25-(OH)D_(3)、ALB、SF可增加学龄前营养不良患儿临床结局不良风险,联合检测对临床结局具有较高预测价值。

关键词：营养不良铁蛋白 25-(OH)D_(3) 白蛋白儿科营养不良评估筛查工具学龄前儿童

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基于区块链技术的高校财务共享模式探究

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中阿科技论坛（中英文） 2023年第11期 80-84页

作者：黄林娟莆田学院审计室福建莆田351100

新政府会计制度在高校的全面实施和“双一流”建设总体方案的持续推进,为高校财务共享服务模式的创新变革提供了良好发展契机。以信息共享、可追溯性为特征的区块链技术近年来快速发展,使得财务共享服务的智能化优化得到了更可靠的技术... 详细信息

新政府会计制度在高校的全面实施和“双一流”建设总体方案的持续推进,为高校财务共享服务模式的创新变革提供了良好发展契机。以信息共享、可追溯性为特征的区块链技术近年来快速发展,使得财务共享服务的智能化优化得到了更可靠的技术支持。文章以高校财务工作为例,基于区块链技术和财务共享模式的基本特征,分析了高校财务共享模式引入区块链技术的必要性,并从财务平台底层应用、财务系统构建以及去中心化理念实施等方面具体探讨了财务共享服务组织架构完善思路以及相关功能调整优化的路径。

关键词：区块链技术高校管理财务共享模式

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