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重组类人胶原蛋白工程菌补料发酵过程建模

中国生物工程杂志 2006年第4期26卷 65-69页

作者：米钰花秀夫范代娣张兮西北大学化工学院西安710069

对产类人胶原蛋白的重组大肠杆菌Escherichia coli(***)的批式和分批-补料培养动力学进行了研究。通过检测发酵过程的基质浓度、菌体量和产物浓度,建立了一组反映发酵的动力学模型,并考虑了非工程菌存在的影响,分析了细胞生长、底物消... 详细信息

对产类人胶原蛋白的重组大肠杆菌Escherichia coli(***)的批式和分批-补料培养动力学进行了研究。通过检测发酵过程的基质浓度、菌体量和产物浓度,建立了一组反映发酵的动力学模型,并考虑了非工程菌存在的影响,分析了细胞生长、底物消耗、基因工程产物生成的过程, 结果显示该动力学模型可以很好地拟合发酵过程。

关键词：动力学发酵胶原蛋白

Process Control for Production of Human-like Collagen in Fed-batch Culture of Escherichia coli BL 21

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Chinese Journal of Chemical Engineering 2005年第2期13卷 276-279页

作者：骆艳娥范代娣马晓轩王德伟米钰花秀夫李稳宏 Department of Chemical Engineering Northwest University/Shanxi Source and Application Chemical Engineering Center Xi'an 710069 China

Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.

关键词： human-like collagen recombinant Escherichia coli fed-batch culture nitrogen oscillation oxygenenrichment temperature inducement

Synthesis and fluorescent properties of a novel europium(Ⅲ) complex with terpyridine-capped poly(ethylene glycol)

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Journal of Rare Earths 2012年第7期30卷 705-708页

作者：张红兴花秀夫庹新林王晓工 Department of Chemical Engineering Laboratory for Advanced MaterialsTsinghua University Office of Scientific Research&Develop-ment Tsinghua University

Terpyridine-capped poly（ethylene glycol）（PEG） was synthesized as a novel polymeric ligand and rare earth complex was prepared from Eu（NO3）3 and the telechelic macromolecules. The structure of the complex was characterized by 1H NMR, FT-IR, UV-vis and fluores-cence spectroscopy. Strong fluorescent emission of the complex was observed at 593 and 616 nm upon the excitation of 300 nm. The fluo-rescent emission was quenched upon addition of trace amount of diethyl chlorophosphate （DCP）. And the observed Ksv values for DCP were measured to be 0.568×10^3 and 0.89×10^3 L/mol for quenching at 593 and 616 nm, which indicated the detection limit for DCP was about 0.014 mmol/L. These results showed that the terpyridine-lanthanide complex could be an effective chemosensor with a potential application in the detection of organophosphates.

关键词： Terpyridine poly(ethylene glycol) (PEG) europium complex fluorescent properties organophosphates rare earths

浅析科研院所中试基地建设的必要性——以化工中试基地为例

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中国高校科技 2017年第S2期 70-71页

作者：张虎胡洋花秀夫清华大学科研院北京100084

当前,行业发展面临的挑战更加突出,技术创新能力弱严重制约着化工行业的可持续发展。限制我国化工行业新产品、新工艺产业化的主要因素有化工产业化需要中试、化工园区入园要求等,因此亟待加强化工中试基地的建设,努力探索企业、政府与... 详细信息

当前,行业发展面临的挑战更加突出,技术创新能力弱严重制约着化工行业的可持续发展。限制我国化工行业新产品、新工艺产业化的主要因素有化工产业化需要中试、化工园区入园要求等,因此亟待加强化工中试基地的建设,努力探索企业、政府与科研院所合作的新形式、新机制,从而推进我国科研院所化工方面原创性成果产业化进程,提升中小化工企业的自主创新能力,以期对现代新型化工中试基地创新模式提供理论依据。

关键词：产学研合作化工中试基地科研院所

Kinetics of High Cell Density Fed-batch Culture of Recombinant Escherichia coli Producing Human-like Collagen

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Chinese Journal of Chemical Engineering 2006年第2期14卷 242-247页

作者：花秀夫范代娣骆艳娥张兮施惠娟米钰马晓轩尚龙安赵桂仿 Shaanxi Source and Application Chemical Engineering Center Department of Chemical Engineering Northwest University Xi'an 710069 China State Education Ministry Key Laboratory of Resource Biotechnology in Western China Department of Biology Northwest University Xi'an 710069 China Department of Reproductive Biology Shanghai Institute of Planned Parenthood Research Shanghai 200032 China College of Biological and Chemical Engineering Ningbo Institute of Technology Zhejiang University Ningbo 315100 China

The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded without consideration of cell endogeneous metabolism. The cell lag time, the maximum specific growth rate and Yx/s were determined as 1.75h, 0.65h^-1 and 0.51g·g^-1, respectively. In the fed-batch culture, different specific growth rates were set at （0.15, 0.2, 0.25h^-1） by the method of pseudo-exponential feeding, and the expressions for the specific rate of substrate consumption, the growth kinetics and the product formation kinetics of each phase were obtained. The result shows that the concentrations of cell and product can reach 77.5g·L^-1 and 10.2g·L^-1 respectively. The modal predictions are in good agreement with the experimental data.

关键词： human-like collagen kinetics high density fermentation Escherichia coli

重组大肠杆菌分批-补料培养生产类人胶原蛋白的过程控制

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西北大学学报（自然科学版） 2005年第2期35卷 180-183页

作者：骆艳娥范代娣花秀夫张兮王小刚西北大学化工学院陕西西安710069

目的优化重组大肠杆菌高密度发酵的调控工艺。方法通过考察比生长速率、3种控氧方式以及诱导强度对细胞和类人胶原蛋白产量的影响,确定最佳的发酵工艺。结果比生长速率显著影响类人胶原蛋白的产量,且最适比生长速率为0.15～0.20h-1;在... 详细信息

目的优化重组大肠杆菌高密度发酵的调控工艺。方法通过考察比生长速率、3种控氧方式以及诱导强度对细胞和类人胶原蛋白产量的影响,确定最佳的发酵工艺。结果比生长速率显著影响类人胶原蛋白的产量,且最适比生长速率为0.15～0.20h-1;在提高罐压的条件下,细胞和类人胶原蛋白的浓度均显著高于其他两种控氧方式,充入富氧空气的细胞和类人胶原蛋白的浓度均最低;诱导强度对细胞生长和蛋白质表达的影响非常大,当细胞浓度达47±2g/L(DCW)时,开始升温至42℃进行诱导,最佳条件为42℃保温2～3h,然后降温至39℃继续培养5～6h。结论通过优化重组大肠杆菌分批补料培养生产类人胶原蛋白的工艺,使得细胞和类人胶原蛋白浓度分别达68.94g/L(DCW)和13.02g/L。

关键词：类人胶原蛋白重组大肠杆菌半分批.补料培养比生长速率控氧方式温度诱导

Polystyrene microsphere-based lanthanide luminescent chemosensor for detection of organophosphate pesticides

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Journal of Rare Earths 2012年第12期30卷 1203-1207页

作者：张红兴花秀夫庹新林陈传治王晓工 Laboratory for Advanced Materials Department of Chemical EngineeringTsinghua University Office of Scientific Research&Develop-ment Tsinghua University Department of Chemical Defense Institute of Chemical Defense

Fluorescent microspheres of polystyrene-based Eu（Ⅲ） complexes were prepared from TentaGel resin, 2,6-bisbenzimidazolylpyridine and europium nitrate. The microspheres were characterized by FTIR spectroscopy, elemental analysis, XPS measurements and fluorescence spectroscopy. Characteristic red emission under irradiation of 365 nm light from a hand-held UV lamp was observed for the microspheres either in a solution or solid-state. Fluorescent quenching was observed when the microspheres were exposed to a trace amount of diethyl chlorophosphate （DCP） in the dispersion. The material and property can be potentially used to fabricate chemosensor in the detection of organophosphates.

关键词： Eu(Ⅲ) complex 2,6-bisbenzimidazolylpyridine TentaGel resin organophosphates fluorescent microbeads rare earths

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石油污染土壤的生物修复技术

生物产业技术 2008年第4期 32-35页

作者：刘铮张坤花秀夫王君韩慧龙清华大学化工系北京100084

简要综述了土壤生物修复技术及其在石油污染土壤修复中的应用现状，着重讨论了生物强化修复技术及其应用过程中所涉及的关键技术问题；介绍了真菌-细菌协同修复石油污染耕地以及该技术与秸秆填埋发酵相集成修复油-盐混合污染耕地的研究... 详细信息

简要综述了土壤生物修复技术及其在石油污染土壤修复中的应用现状，着重讨论了生物强化修复技术及其应用过程中所涉及的关键技术问题；介绍了真菌-细菌协同修复石油污染耕地以及该技术与秸秆填埋发酵相集成修复油-盐混合污染耕地的研究成果。

关键词：土壤修复生物强化修复石油污染土壤

构筑全方位多层次的产学研合作新模式——清华大学与无锡市合作介绍

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中国高校科技 2017年第S2期 106-107页

作者：张虎谢旭东花秀夫胡洋清华大学科技开发部北京100084

自清华大学与无锡市人民政府于2005年3月签署全面合作协议,成立清华大学—无锡产学研合作办公室以来,双方在科技项目对接、企业需求调研、企业现场诊断、国际科技交流、人才进修培训、毕业学生招聘、毕业学生挂职、设立实践基地等多方... 详细信息

自清华大学与无锡市人民政府于2005年3月签署全面合作协议,成立清华大学—无锡产学研合作办公室以来,双方在科技项目对接、企业需求调研、企业现场诊断、国际科技交流、人才进修培训、毕业学生招聘、毕业学生挂职、设立实践基地等多方面实现了卓有成效的合作。尤其是近年来,双方围绕共建科研机构、开展多层次的全方位深度科技合作开展了有益的探索,取得了良好的效果。

关键词：校地科技合作产学研合作科技创新