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过表达果糖脱氢酶促进氧化葡萄糖酸杆菌高效合成5-酮果糖

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食品与生物技术学报 2024年第6期43卷 58-66页

作者：钱雪赵晨休林金萍魏东芝华东理工大学生物工程学院上海200237 华东理工大学生物反应器工程国家重点实验室上海200237 华东理工大学中国轻工业生物催化与智能制造重点实验室上海200237

5-酮果糖(5-KetoFructose,5-KF)具有与D-果糖相似的甜度和自然甜味,且不能被人体代谢而具有低热量的优点,是一种具有发展前景的非营养型天然甜味剂。为了获得5-酮果糖高产菌株,作者将来源于Gluconobacter japonicus的膜结合的果糖脱氢... 详细信息

5-酮果糖(5-KetoFructose,5-KF)具有与D-果糖相似的甜度和自然甜味,且不能被人体代谢而具有低热量的优点,是一种具有发展前景的非营养型天然甜味剂。为了获得5-酮果糖高产菌株,作者将来源于Gluconobacter japonicus的膜结合的果糖脱氢酶基因在Gluconobacter oxydans中过表达,通过对不同的宿主、表达载体和启动子的筛选,获得一株能够高效合成5-KF的工程菌QT-10。在3 L发酵罐中进行补料发酵优化,最终采用恒速连续流加果糖的方式,该工程菌可在82 h内消耗质量浓度为650 g/L的D-果糖,5-KF的产量高达608 g/L,产率为94.6%,时空产率为7.41 g/(L·h)。

关键词： 5-酮果糖果糖脱氢酶氧化葡萄糖酸杆菌重组表达发酵优化

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“酶工程”国家一流本科课程的建设与探索

生物工程学报 2024年第3期40卷 943-952页

作者：王华磊高伟杰许萌林金萍王学东魏东芝华东理工大学生物工程学院上海200237

生物技术产业是我国战略性新兴产业,在整个国民经济中占有重要地位,创新型的高素质专业人才的培养意义重大。“酶工程”作为生物技术专业核心课程,教学质量的好坏直接影响学生质量及行业发展。本课程针对目前课程教学中存在的课程内容... 详细信息

生物技术产业是我国战略性新兴产业,在整个国民经济中占有重要地位,创新型的高素质专业人才的培养意义重大。“酶工程”作为生物技术专业核心课程,教学质量的好坏直接影响学生质量及行业发展。本课程针对目前课程教学中存在的课程内容优化不足、过分依赖教材、教学方法单一等问题,以调整大纲、更新案例素材库、调动学生积极性为抓手,通过立体式教学-线上线下结合、互动式教学-翻转课堂、启发式教学-基于问题的学习(problem-based learning,PBL)教学模式、专题式教学-案例教学和完善考核机制等多种手段,激发学生的学习热情与创新能力,以培养厚基础、强实践的高素质专业人才。通过教学探索与实践,教学效果改善明显,学生的工程实践能力和创新能力得到了极大程度的提高。

关键词：酶工程生物技术教学内容教学方法

A CRISPRi mediated self-inducible system for dynamic regulation of TCA cycle and improvement of itaconic acid production in Escherichia coli

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Synthetic and Systems Biotechnology 2022年第3期7卷 982-988页

作者： Ming Zhao Yuting Li Fengqing Wang Yuhong Ren dongzhi wei State Key Lab of Bioreactor Engineering Newworld Institute of BiotechnologyEast China University of Science and TechnologyShanghai200237China Anhui Engineering Laboratory for Industrial Microbiology Molecular Breeding College of Biology and Food EngineeringAnhui Polytechnic UniversityWuhu241000China

Itaconic acid(ITA),an effective alternative fossil fuel,derives from the bypass pathway of the tricarboxylic acid(TCA)***,the imbalance of metabolic flux between TCA cycle and ITA biosynthetic pathway seriously limits the production of *** optimization of flux distribution between biomass and production has the potential to the productivity of *** on the previously constructed strain Escherichia coli MG1655Δ1-SAS-3(ITA titer:1.87 g/L),a CRISPRi-mediated self-inducible system(CiMS),which contained a responsive module based on the ITA biosensor YpItcR/Pccl and a regulative CRISPRi-mediated interferential module,was developed to regulate the flux of the TCA cycle and to enhance the capacity of the strain to produce ***,a higher ITA-yielding strain,Δ4-Prmd-SAS-3(ITA titer:3.20 g/L),derived fromΔ1-SAS-3,was constructed by replacing the promoter PJ23100,for the expression of ITA synthesis genes,with Prmd and knocking out the three bypass genes poxB,pflB,and ***,the CiMS was used to inhibit the expression of key genes icd,pykA,and sucCD to dynamically balance the metabolic flux between TCA cycle and ITA biosynthetic pathway during the ITA production *** constructed strainΔ4-Prmd-SAS-3 under the dynamic regulation of the CiMS,showed a 23%increase in the ITA titer,which reached 3.93 g/*** study indicated that CiMS was a practical strategy to dynamically and precisely regulated the metabolic flux in microbial cell factories.

关键词： CRISPRi Dynamic regulation Itaconic acid Biosensor

A screening analysis of the GJB2 c.176 del 16 mutation responsible for hereditary deafness in a Chinese family

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Journal of Otology 2016年第3期11卷 134-137页

作者： Hao Jiang Xi Shi Shiwei Qiu Yanfen Dong Yuehua Qiao dongzhi wei New World Institute of Biotechnology State Key Laboratory of Bioreactor EngineeringEast China University of Science and Technology The Institute of Audiology and Balance Science of Xuzhou Medical University Clinical Hearing Center of Affiliated Hospital of Xuzhou Medical University

Objective:To determine whether a new-born child from a family carrying a deafness gene needs cochlear implantation to avoid dysphonia by screening and sequencing a deafness-related ***:Both screening and sequencing results confirmed that the new born child had a normal GJB2 gene despite the fact that she has a brother suffering from hearing loss triggered by an allelic GJB2 c.176 del 16 *** cloned the GJB2 genes derived from their respective blood genomic DNA into GFP fused plasmids and transfected those plasmids into the 293 T cell line to test for gene *** the mutated GJB2gene(GJB2 c.176 del 16) of her deaf brother was found to be unable to form the gap junction structure between two adjacent cells,the baby girl’s GJB2 gene ran into no such ***:The screening and sequencing as well as the GJB2 gene function tests invariably showed results consistent with the ABR tested hearing phenotype,which means that the child,with a normal wild type GJB2 gene,does not need early intervention to prevent her from developing hearing loss and dysphonia at a later stage in life.

关键词： Deafness gene screening GJB2 Gap junction

COMPLEX REACTIONS OF ETHYL-GLUCOSIDES SYNTHESIS OVER ION EXCHANGE RESIN

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化工学报 2000年第S1期51卷 163-167页

作者： Tu Maobing and wei dongzhi (The State Key Laboratory of Bioreactor Engineering ECUST, Shanghai 200237)

The glycosidation reactions of D-glucose with ethanol have been carried out over a reusable and separable heterogeneous catalyst, namely, ion exchange resin. Detailed kinetic data for these reactions are reported. A complex reaction model has been developed for interpreting the data. The reactions were found to be global second-order reactions and first -order with respect to each component. Meanwhile, a new regression method is applied to determine the rate constant from time-dependent profiles.

关键词： Ethyl-glucoside solid catalyst kinetic complex reactions

定制酶分子机器/细胞工厂,引领生物制造产业未来——2017年第十一届中国酶工程学术研讨会杜邦-杰能科中国酶工程杰出贡献奖获得者特邀报告

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生物工程学报 2018年第7期34卷 1024-1032页

作者：姜水琴魏东芝华东理工大学生物工程学院生物反应器工程国家重点实验室鲁华生物技术研究所上海200237

工业酶制剂研发与应用已经渗透到各大工业领域,但中国作为用酶大国、产酶小国面临重大挑战,鉴于以化学催化为核心的基础物质加工业面临资源、能源和环境三大危机,酶工程与生物催化已被列入许多国家的科技与产业发展战略,应用高效、清洁... 详细信息

工业酶制剂研发与应用已经渗透到各大工业领域,但中国作为用酶大国、产酶小国面临重大挑战,鉴于以化学催化为核心的基础物质加工业面临资源、能源和环境三大危机,酶工程与生物催化已被列入许多国家的科技与产业发展战略,应用高效、清洁的生物催化技术是实现化学工业可持续发展以及发酵工业产业升级的重要途径之一。文中以2017年第十一届中国酶工程学术研讨会杜邦-杰能科中国酶工程杰出贡献奖获得者特邀报告为基础整理编写而成,从自主酶库构建、酶分子机器/细胞工厂创制及产业化应用等角度概述当前酶工程与生物催化发展现状及前景。

关键词：酶分子机器细胞工厂生物制造产业未来催化剂定制

分枝杆菌Mycobacterium ***-01 3-甾酮-9α-羟基化酶基因的克隆、异源表达及分离纯化

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生物工程学报 2009年第12期25卷 2014-2021页

作者：范书玥魏巍王风清魏东芝华东理工大学生物反应器工程国家重点实验室鲁华生物技术研究所上海200237

3-甾酮-9α-羟基化酶(KSH)是微生物甾体降解途径中的关键酶,在甾体药物制备中有重要价值。以本实验室从土壤中自行筛选的分枝杆菌Mycobacterium ***-01为出发菌株,利用红平红球菌Rhodococcus erythropolis SQ1已报道的ksh序列与已全基... 详细信息

3-甾酮-9α-羟基化酶(KSH)是微生物甾体降解途径中的关键酶,在甾体药物制备中有重要价值。以本实验室从土壤中自行筛选的分枝杆菌Mycobacterium ***-01为出发菌株,利用红平红球菌Rhodococcus erythropolis SQ1已报道的ksh序列与已全基因组测序的分枝杆菌序列数据库进行比对分析,根据同源基因设计简并引物获得部分ksh序列,通过染色体步移扩增出全长ksh(命名为M.S.-ksh),该基因与耻垢分枝杆菌*** mc2155的ksh同源性为85%。构建pET32-ksh表达载体,转化大肠杆菌BL21(DE3),获得高表达重组转化子菌株,经IPTG低温诱导,SDS-PAGE电泳分析,目的蛋白主要为可溶性表达,表达量占菌体总蛋白的30%以上,用Ni2+亲和层析柱纯化,纯度达90%以上。本研究为利用基因工程菌进行工业化生产甾体药物奠定了基础。

关键词：分枝杆菌NwIB-01 3-甾酮-9α-羟基化酶甾体降解异源表达分离纯化

A simple fabrication method of germanium-based metasurfaces

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Chinese Optics Letters 2024年第7期22卷 117-120页

作者： dongzhi Shan Jinsong Gao Nianxi Xu Naitao Song Hongda wei Qiang Li Yi Zhao Hai Liu Qiao Sun Xiaonan Jiang Chao Li Zhiyu Zhang Key Laboratory of Optical System Advanced Manufacturing Technology Changchun Institute of OpticsFine Mechanics and PhysicsChinese Academy of SciencesChangchun 130033China State Key Laboratory of Applied Optics Changchun Institute of OpticsFine Mechanics and PhysicsChinese Academy of SciencesChangchun 130033China Jilin Provincial Key Laboratory of Advanced Optoelectronic Equipment and Instrument Manufacturing Technology Changchun 130033China

In this work,a simple fabrication method of germanium-based metasurfaces is proposed,where the deposited Al_(2)O_(3) layer with high selectivity is chosen as the hard mask and retained after the dry etching *** simulation and experimental characterization results verify the feasibility of the fabrication *** experimental study on the fabrication methods of germanium-based metasurfaces is very significant as the meta-atoms with a higher refractive index can achieve 0 to 2πtransmission phase variation with a smaller period under the same thickness-to-period ratio,which is consistent with the requirement of the period miniaturization in some cases.

关键词： metasurfaces period miniaturization refractive index.

汽车用DP800双相钢电阻点焊温度场数值模拟研究

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热加工工艺 2016年第5期45卷 243-246页

作者：张梅红魏冬至河南工业贸易职业学院机电工程系河南郑州450000 郑州市技师学院河南郑州450000

采用SORPAS专用电阻焊有限元软件,对1.4 mm的DP800双相钢电阻点焊温度场进行了模拟,并将熔核形貌模拟结果与实验结果进行对比。结果表明,在参考工艺下,通电加热阶段电极和焊件峰值均升高,工件温度升高速度明显高于电极,通电过程结束时... 详细信息

采用SORPAS专用电阻焊有限元软件,对1.4 mm的DP800双相钢电阻点焊温度场进行了模拟,并将熔核形貌模拟结果与实验结果进行对比。结果表明,在参考工艺下,通电加热阶段电极和焊件峰值均升高,工件温度升高速度明显高于电极,通电过程结束时电极峰值温度达最高值566℃,焊件峰值温度达2120℃;随着焊接电流的增加,热循环曲线逐渐上移,加热速度和最高温度逐渐升高,高温停留时间延长;随着焊接时间的增加,加热速度没有明显变化,最高温度逐渐升高,高温停留时间延长;在没有飞溅的条件下模拟熔核形貌与实验结果基本吻合,但在大电流条件下由于飞溅的产生,造成模拟熔核尺寸大于实际结果。

关键词： DP800双相钢电阻点焊数值模拟