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Exsolved materials for CO_(2)reduction in high-temperature electrolysis cells 被引量:1
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作者 Min Xu Ran Cao +5 位作者 Han Qin Nuoxi Zhang Wenle Yan liming liu John T.S.Irvine Di Chen 《Materials Reports(Energy)》 2023年第2期62-81,I0003,共21页
Electrochemical reduction of CO_(2)into valuable fuels and chemicals has become a contemporary research area,where the heterogeneous catalyst plays a critical role.Metal nanoparticles supported on oxides performing as... Electrochemical reduction of CO_(2)into valuable fuels and chemicals has become a contemporary research area,where the heterogeneous catalyst plays a critical role.Metal nanoparticles supported on oxides performing as active sites of electrochemical reactions have been the focus of intensive investigation.Here,we review the CO_(2)reduction with active materials prepared by exsolution.The fundamental of exsolution was summarized in terms of mechanism and models,materials,and driven forces.The advances in the exsolved materials used in hightemperature CO_(2)electrolysis were catalogued into tailored interfaces,synergistic effects on alloy particles,phase transition,reversibility and electrochemical switching. 展开更多
关键词 CO_(2)reduction EXSOLUTION Solid oxide electrolysis cells CATALYSTS
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Concentration-Dependent Effect of Nickel Ions on Amyloid Fibril Formation Kinetics of Hen Egg White Lysozyme:a Raman Spectroscopy Study
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作者 Xinfei Li Xiaodong Chen +3 位作者 Ning Chen liming liu Xiaoguo Zhou Shilin liu 《Chinese Journal of Chemical Physics》 SCIE EI CAS CSCD 2023年第5期517-525,I0001,共10页
Nickel,an important transi-tion metal element,is one of the trace elements for hu-man body and has a crucial impact on life and health.Some evidences show the excess exposure to metal ions might be associated with neu... Nickel,an important transi-tion metal element,is one of the trace elements for hu-man body and has a crucial impact on life and health.Some evidences show the excess exposure to metal ions might be associated with neurological diseases.Herein,we applied Raman spectroscopy to study the Ni(II)ion effect on kinetics of amyloid fibrillation of hen egg white lysozyme(HEWL)in thermal and acidic conditions.Using the well-known Raman indicators for protein tertiary and secondary structures,we monitored and analyzed the concentration effect of Ni(II)ions on the unfolding of tertiary structures and the transformation of sec-ondary structures.The experimental evidence validates the accelerator role of the metal ion in the kinetics.Notably,the additional analysis of the amide I band profile,combined with thioflavin-T fluorescence assays,clearly indicates the inhibitory effect of Ni(II)ions on the formation of amyloid fibrils with organizedβ-sheets structures.Instead,a more significant promotion influence is affirmed on the assembly into other aggregates with disordered struc-tures.The present results provide rich information about the specific metal-mediated protein fibrillation. 展开更多
关键词 Amyloid fibrillation Protein denaturation KINETICS Nickel ion LYSOZYME
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Hole‑Transport Management Enables 23%‑Efficient and Stable Inverted Perovskite Solar Cells with 84%Fill Factor
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作者 liming liu Yajie Ma +7 位作者 Yousheng Wang Qiaoyan Ma Zixuan Wang Zigan Yang Meixiu Wan Tahmineh Mahmoudi Yoon‑Bong Hahn Yaohua Mai 《Nano-Micro Letters》 SCIE EI CAS CSCD 2023年第8期154-166,共13页
NiO_(x)-based inverted perovskite solar cells(PSCs)havepresented great potential toward low-cost,highly efficient and stablenext-generation photovoltaics.However,the presence of energy-levelmismatch and contact-interf... NiO_(x)-based inverted perovskite solar cells(PSCs)havepresented great potential toward low-cost,highly efficient and stablenext-generation photovoltaics.However,the presence of energy-levelmismatch and contact-interface defects between hole-selective contacts(HSCs)and perovskite-active layer(PAL)still limits device efficiencyimprovement.Here,we report a graded configuration based on bothinterface-cascaded structures and p-type molecule-doped compositeswith two-/three-dimensional formamidinium-based triple-halideperovskites.We find that the interface defects-induced non-radiativerecombination presented at HSCs/PAL interfaces is remarkably suppressedbecause of efficient hole extraction and transport.Moreover,astrong chemical interaction,halogen bonding and coordination bondingare found in the molecule-doped perovskite composites,whichsignificantly suppress the formation of halide vacancy and parasitic metallic lead.As a result,NiO_(x)-based inverted PSCs present a power-conversion-efficiency over 23%with a high fill factor of 0.84 and open-circuit voltage of 1.162 V,which are comparable to the best reported around 1.56-electron volt bandgap perovskites.Furthermore,devices with encapsulation present high operational stability over 1,200 h during T_(90) lifetime measurement(the time as a function of PCE decreases to 90%of its initial value)under 1-sun illumination in ambient-air conditions. 展开更多
关键词 Inverted NiO_(x)-based perovskite solar cells Hole-transport management Interface-induced defect passivation High performance and stability
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Distributed Virtual Inertia Based Control of Multiple Photovoltaic Systems in Autonomous Microgrid 被引量:3
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作者 Won-Sang Im Cheng Wang +2 位作者 Wenxin liu liming liu Jang-Mok Kim 《IEEE/CAA Journal of Automatica Sinica》 SCIE EI CSCD 2017年第3期512-519,共8页
The large inertia of a traditional power system slows down system's frequency response but also allows decent time for controlling the system.Since an autonomous renewable microgrid usually has much smaller inerti... The large inertia of a traditional power system slows down system's frequency response but also allows decent time for controlling the system.Since an autonomous renewable microgrid usually has much smaller inertia,the control system must be very fast and accurate to fight against the small inertia and uncertainties.To reduce the demanding requirements on control,this paper proposes to increase the inertia of photovoltaic(PV) system through inertia emulation.The inertia emulation is realized by controlling the charging/discharging of the direct current(DC)-link capacitor over a certain range and adjusting the PV generation when it is feasible and/or necessary.By well designing the inertia,the DC-link capacitor parameters and the control range,the negative impact of inertia emulation on energy efficiency can be reduced.The proposed algorithm can be integrated with distributed generation setting algorithms to improve dynamic performance and lower implementation requirements.Simulation studies demonstrate the effectiveness of the proposed solution. 展开更多
关键词 Inertia emulation MICROGRID photovoltaic system renewable energy voltage source converter
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Ultrasonic Welding of Magnesium–Titanium Dissimilar Metals:A Study on Thermo-mechanical Analyses of Welding Process by Experimentation and Finite Element Method 被引量:2
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作者 Dewang Zhao Daxin Ren +3 位作者 Kunmin Zhao Pan Sun Xinglin Guo liming liu 《Chinese Journal of Mechanical Engineering》 SCIE EI CAS CSCD 2019年第6期181-191,共11页
Ultrasonic welding is an effective ways to achieve a non-reactive/immiscible heterogeneous metal connection, such as the connection of magnesium alloy and titanium alloy. But the thermal mechanism of magnesium alloy/t... Ultrasonic welding is an effective ways to achieve a non-reactive/immiscible heterogeneous metal connection, such as the connection of magnesium alloy and titanium alloy. But the thermal mechanism of magnesium alloy/titanium alloy ultrasonic welding has not been defined clearly. In this paper, the experimental and the finite element analysis were adopted to study the thermal mechanism during welding. Through the test, the temperature variation law during the welding process is obtained, and the accuracy of the finite element model is verified. The microscopic analysis indicates that at the welding time of 0.5 s, the magnesium alloy in the center of the solder joint is partially melted and generates the liquid phase. Through the finite element analysis, the friction coefficient of the magnesium–titanium ultrasonic welding interface can be considered as an average constant value of 0.28. The maximum temperature at the interface can exceed 600 ℃ to reach the melting point temperature of the magnesium alloy. The plastic deformation begins after 0.35 s and occurs at the magnesium side at the center of the interface. 展开更多
关键词 Ultrasonic welding Magnesium alloys Titanium alloys Thermo-mechanical analyses Finite elementanalysis
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Enhancement of α-ketoisovalerate production by relieving the product inhibition of L-amino acid deaminase from Proteus mirabilis 被引量:2
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作者 Shanshan Pei Xiaobo Ruan +5 位作者 Jia liu Wei Song Xiulai Chen Qiuling Luo liming liu Jing Wu 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2020年第8期2190-2199,共10页
L-Amino acid deaminase(LAAD) is a key enzyme in the deamination of L-valine(L-val) to produce α-ketoisovalerate(KIV). However, the product inhibition of LAAD is a major hindrance to industrial KIV production.In the p... L-Amino acid deaminase(LAAD) is a key enzyme in the deamination of L-valine(L-val) to produce α-ketoisovalerate(KIV). However, the product inhibition of LAAD is a major hindrance to industrial KIV production.In the present study, a combination strategy of modification of flexible loop regions around the product binding site and the avoidance of dramatic change of main-chain dynamics was reported to reduce the product inhibition.The four mutant PM-LAAD^(M4)(PM-LAAD^(S98A/T105A/S106A/L341A)) achieved a 6.2-fold higher catalytic efficiency and an almost 6.7-fold reduction in product inhibition than the wild-type enzyme. Docking experiments suggested that weakened interactions between the product and enzyme, and the flexibility of the "lid" structure relieved LAAD product inhibition. Finally, the whole-cell biocatalyst PM-LAAD^(M4) has been applied to KIV production,the titer and conversion rate of KIV from L-val were 98.5 g·L^-1 and 99.2% at a 3-L scale, respectively. These results demonstrate that the newly engineered catalyst can significantly reduce the product inhibition, that making KIV a prospective product by bioconversion method, and also provide the understanding of the mechanism of the relieved product inhibition of PM-LAAD. 展开更多
关键词 Product inhibition L-Amino acid deaminases α-Ketoisovalerate Bio-catalysis Protein engineering
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Preparation and performance analysis of a coking coal dust suppressant spray 被引量:1
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作者 Jiuhong Jiang Pengfei Wang +3 位作者 Ye Pei Ronghua liu liming liu Yunchu He 《International Journal of Coal Science & Technology》 EI CAS CSCD 2021年第5期1003-1014,共12页
Coking coal dust is extremely hydrophobic;therefore,combination with droplets in the air is difficult and dust suppression is challenging.Here,a dust suppressant spray for coking coal dust was studied in order to impr... Coking coal dust is extremely hydrophobic;therefore,combination with droplets in the air is difficult and dust suppression is challenging.Here,a dust suppressant spray for coking coal dust was studied in order to improve of the combination of droplets and coking coal dust.Based on monomer optimization and compounding analysis,two surfactant monomers,fatty alcohol ether sodium sulfate(AES)and sodium dodecyl benzene sulfonate(SDBS)were selected as the surfactant components of the dust suppressant.The surfactant monomers were combined with four inorganic salts and the reverse osmosis moisture absorption of each solution was determined.By combining the reverse osmosis moisture absorption values with the water retention experimental results,CaCl_(2)was identified as the optimal inorganic salt additive for the dust suppressant.Finally,the optimal concentration of each component was obtained using orthogonal experimental design i.e.,AES(0.03%),SDBS(0.05%),and CaCl_(2)(0.4%).The dust suppressant solution formulated using this method had a high moisture absorption capacity and excellent performance. 展开更多
关键词 Coking coal DUST SPRAY Dust suppressant SURFACTANT Inorganic salt additive
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Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli 被引量:1
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作者 Xiaobo Ruan Sheng Zhang +4 位作者 Wei Song Jia liu Xiulai Chen liming liu Jing Wu 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2022年第7期18-30,共13页
For the efficient conversion of L-tyrosine(L-Tyr)to tyrosol,which is an aromatic compound widely used in the pharmaceutical and chemical industries,a novel four-enzyme cascade pathway based on the Ehrlich pathway of S... For the efficient conversion of L-tyrosine(L-Tyr)to tyrosol,which is an aromatic compound widely used in the pharmaceutical and chemical industries,a novel four-enzyme cascade pathway based on the Ehrlich pathway of Saccharomyces cerevisiae was designed and reconstructed in Escherichia coli.Then,the expression levels of the relevant enzymes were coordinated using a modular approach and gene duplication after the identification of the pyruvate decarboxylase from Candida tropicalis(CtPDC)as the rate-limiting enzymatic step.In situ product removal(ISPR)strategy with XAD4 resins was explored to avoid product inhibition and further improve tyrosol yield.As a result,the titer and conversion rate of tyrosol obtained were 35.7 g·L^(-1) and 93.6%,respectively,in a 3-L bioreactor.Results presented here provide a potential enzymatic process for industrial production of tyrosol from cheap amino acids. 展开更多
关键词 TYROSOL L-TYROSINE Ehrlich pathway Enzyme cascade In situ product removal
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Enhanced spin-dependent thermopower in a double-quantum-dot sandwiched between two-dimensional electron gases
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作者 迟锋 付振国 +1 位作者 刘黎明 张平 《Chinese Physics B》 SCIE EI CAS CSCD 2019年第10期523-530,共8页
We study the spin-dependent thermopower in a double-quantum-dot(DQD) embedded between the left and right two-dimensional electron gases(2DEGs) in doped quantum wells under an in-plane magnetic field. When the separati... We study the spin-dependent thermopower in a double-quantum-dot(DQD) embedded between the left and right two-dimensional electron gases(2DEGs) in doped quantum wells under an in-plane magnetic field. When the separation between the DQD is smaller than the Fermi wavelength in the 2DEGs, the asymmetry in the dots' energy levels leads to pronounced quantum interference effects characterized by the Dicke line-shape of the conductance, which are sensitive to the properties of the 2DEGs. The magnitude of the thermopower, which denotes the generated voltage in response to an infinitesimal temperature difference between the two 2DEGs under vanishing charge current, will be obviously enhanced by the Dicke effect. The application of the in-plane magnetic field results in the polarization of the spin-up and spin-down conductances and thermopowers, and enables an efficient spin-filter device in addition to a tunable pure spin thermopower in the absence of its charge counterpart. 展开更多
关键词 quantum DOT spin-dependent THERMOPOWER TWO-DIMENSIONAL ELECTRON GASES
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Two-dimensional hexagonal Zn3Si2 monolayer:Dirac cone material and Dirac half-metallic manipulation
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作者 官雨柔 宋玲玲 +4 位作者 赵慧 杜仁君 刘力铭 闫翠霞 蔡金明 《Chinese Physics B》 SCIE EI CAS CSCD 2020年第8期418-423,共6页
The fascinating Dirac cone in honeycomb graphene,which underlies many unique electronic properties,has inspired the vast endeavors on pursuing new two-dimensional(2D)Dirac materials.Based on the density functional the... The fascinating Dirac cone in honeycomb graphene,which underlies many unique electronic properties,has inspired the vast endeavors on pursuing new two-dimensional(2D)Dirac materials.Based on the density functional theory method,a 2D material Zn3Si2 of honeycomb transition-metal silicide with intrinsic Dirac cones has been predicted.The Zn3Si2 monolayer is dynamically and thermodynamically stable under ambient conditions.Importantly,the Zn3Si2 monolayer is a room-temperature 2D Dirac material with a spin-orbit coupling energy gap of 1.2 meV,which has an intrinsic Dirac cone arising from the special hexagonal lattice structure.Hole doping leads to the spin polarization of the electron,which results in a Dirac half-metal feature with single-spin Dirac fermion.This novel stable 2D transition-metal-silicon-framework material holds promises for electronic device applications in spintronics. 展开更多
关键词 two-dimensional(2D)Dirac cone material Dirac half-metal first-principles calculation spin-orbit coupling
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Risk Factors for Prognosis after the Maze Ⅳ Procedure in Patients with Atrial Fibrillation Undergoing valve Surgery
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作者 Hui Yang Pu Zou +5 位作者 Yuhu He Lingzhi Huang Xinmin Zhou liming liu Zhenjiang liu Shenghua Zhou 《Cardiovascular Innovations and Applications》 2022年第1期135-145,共11页
The present study evaluated risk factors related to persistent atrial fibrillation(AF)at discharge(AF-d)and recurrent atrial fibrillation(rAF)and all-cause death after the maze IV procedure.Two hundred nineteen patien... The present study evaluated risk factors related to persistent atrial fibrillation(AF)at discharge(AF-d)and recurrent atrial fibrillation(rAF)and all-cause death after the maze IV procedure.Two hundred nineteen patients(63 female,aged 52.5±8.8 years)with valve disease and persistent AF undergoing valve surgery and the maze IV procedure in our center between 2015 and 2016 were included.Baseline demographic and clinical data were obtained by review of medical records.The median follow-up period was 27 months(interquartile range 21-34 months)in our patient cohort.The primary end point was all-cause death.The secondary end point was AF-d or rAF.rAF is defined as AF recurrence at 3 months or later after the procedure.Twenty-eight patients(12.8%)died during follow-up.Multiple logistic regression analysis showed that thrombocytopenia,elevated serum total bilirubin level,a larger right atrium,AF-d,and rAF were independent determinants for all-cause death after the maze IV procedure after adjustment for age,sex,and clinical covariates,including New York Heart Association class III/IV disease,hypertension,and aortic regurgitation,while valvular disease duration and left atrial diameter greater than 80.5 mm were independent determinants for AF-d,and thrombocytopenia,elevated serum total bilirubin level,higher mean pulmonary artery pressure,and AF-d were independent predictors for rAF.In conclusion,thrombocytopenia,elevated serum total bilirubin level,an enlarged right atrium,AF-d,and rAF are independent predictors of all-cause death in patients undergoing the maze IV procedure. 展开更多
关键词 Atrial fibrillation maze IV valve disease THROMBOCYTOPENIA
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Photoacoustic molecular imaging with functional nanoparticles
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作者 liming liu Huan Qin 《Journal of Innovative Optical Health Sciences》 SCIE EI CAS 2017年第4期17-28,共12页
Photoacoustic imaging(PAI)breaks through the optical di®usion limit by making use of the PA e®ect.By converting incident photons into ultrasonic waves,PAI combines high contrast of optical imaging and high s... Photoacoustic imaging(PAI)breaks through the optical di®usion limit by making use of the PA e®ect.By converting incident photons into ultrasonic waves,PAI combines high contrast of optical imaging and high spatial resolution in depth tissue of ultrasound imaging in a single imaging modality.This imaging modality has now shown potential for molecular imaging,which enables visualization of biological processes with systemically introduced functional nanoparticles.In the current review,the potentials of di®erent optical nanoprobes as PAI contrast agents were elucidated and discussed. 展开更多
关键词 Photoacoustic imaging optical nanoprobes molecular imaging
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Metabolic engineering strategies for microbial utilization of C1 feedstocks
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作者 Jian Zhang Liang Guo +4 位作者 Cong Gao Wei Song Jing Wu liming liu Xiulai Chen 《Systems Microbiology and Biomanufacturing》 2023年第1期122-136,共15页
The use of abundant and cheap one carbon(C1)feedstocks to produce value-added chemicals is an important approach for achieving carbon neutrality and tackling environmental problems.The conversion of C1 feedstocks to h... The use of abundant and cheap one carbon(C1)feedstocks to produce value-added chemicals is an important approach for achieving carbon neutrality and tackling environmental problems.The conversion of C1 feedstocks to high-value chemicals is dependent on efficient C1 assimilation pathways and microbial chassis adapted for efficient incorporation.Here,we opted to summarize the natural and synthetic C1 assimilation pathways and their key factors for metabolizing C1 feedstock.Accordingly,we discussed the metabolic engineering strategies for enabling the microbial utilization of C1 feedstocks for the bioproduction of value-added chemicals.In addition,we highlighted future perspectives of C1-based biomanufacturing for achieving a low-carbon footprint for the biosynthesis of chemicals. 展开更多
关键词 C1-based biomanufacturing C1 assimilation pathways Metabolic engineering strategies C1 feedstocks Chemicals biosynthesis
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Advances in microbial engineering for the production of value‑added products in a biorefinery
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作者 Cong Gao Liang Guo +3 位作者 Wei Song Jing Wu Xiulai Chen liming liu 《Systems Microbiology and Biomanufacturing》 2023年第2期246-261,共16页
Microbial biorefineries to produce chemicals from renewable feedstock provides attractive advantages,including mild reaction conditions and sustainable manufacturing.However,low-efficiency biorefineries always result ... Microbial biorefineries to produce chemicals from renewable feedstock provides attractive advantages,including mild reaction conditions and sustainable manufacturing.However,low-efficiency biorefineries always result in an uncompetitive biological process compared to the current petrochemical process.Thus,improving microbial capacity to maximize product yield,productivity,and titer has been recognized as a central goal for bioengineers and biochemists.The knowledge of cellular biochemistry has enabled the regulation of microbial physiology to couple with chemical production.The rapid development in metabolic engineering provides diverse strategies to enhance the efficiency of chemical biosynthesis pathways.New synthetic biology tools as well as novel regulatory targets also offer the opportunity to improve biorefinery environmental adaptivity.In this review,the recent advances in building efficient biorefineries were showcased.In addition,the challenges and future perspectives of microbial host engineering for increased microbial capacity of a biorefinery were discussed. 展开更多
关键词 Microbial cell factory Metabolic engineering Synthetic biology BIOREFINERY
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Metabolic engineering strategies for microbial utilization of methanol
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作者 Yamei Gan Xin Meng +3 位作者 Cong Gao Wei Song liming liu Xiulai Chen 《Engineering Microbiology》 2023年第3期37-50,共14页
The increasing shortage of fossil resources and environmental pollution has renewed interest in the synthesis of value-added biochemicals from methanol.However,most of native or synthetic methylotrophs are unable to a... The increasing shortage of fossil resources and environmental pollution has renewed interest in the synthesis of value-added biochemicals from methanol.However,most of native or synthetic methylotrophs are unable to assimilate methanol at a sufficient rate to produce biochemicals.Thus,the performance of methylotrophs still needs to be optimized to meet the demands of industrial applications.In this review,we provide an in-depth discussion on the properties of natural and synthetic methylotrophs,and summarize the natural and synthetic methanol assimilation pathways.Further,we discuss metabolic engineering strategies for enabling microbial utilization of methanol for the bioproduction of value-added chemicals.Finally,we highlight the potential of microbial engineering for methanol assimilation and offer guidance for achieving a low-carbon footprint for the biosynthesis of chemicals. 展开更多
关键词 METHANOL METHYLOTROPHS Methanol assimilation pathways Metabolic engineering strategies
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支持向量机的关键问题和展望 被引量:12
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作者 邵元海 刘黎明 +1 位作者 黄凌伟 邓乃扬 《中国科学:数学》 CSCD 北大核心 2020年第9期1233-1248,共16页
作为机器学习的主要方法之一,支持向量机不仅有坚实的统计学习理论基础,而且在众多领域中表现出优秀的泛化性能,因此受到了广泛关注.然而近几年来,相比于深度学习的蓬勃发展,支持向量机的研究进展缓慢.本文从支持向量机的本质出发,探讨... 作为机器学习的主要方法之一,支持向量机不仅有坚实的统计学习理论基础,而且在众多领域中表现出优秀的泛化性能,因此受到了广泛关注.然而近几年来,相比于深度学习的蓬勃发展,支持向量机的研究进展缓慢.本文从支持向量机的本质出发,探讨支持向量机的理论方法与深度学习等机器学习热点研究的交叉与融合,提出一些新的思路.具体地,包括3个方面:支持向量机的大间隔原则及其带来的低密度性、核映射的高维划分技巧及其统计学习理论,以及支持向量机的浅层学习模式向深度学习和广度学习的拓展.同时,从这3个方面分别提出支持向量机研究中可以进一步挖掘的优良性质,并展望未来可能诱导出的理论和方法. 展开更多
关键词 支持向量机 统计学习 核学习 机器学习 最优化 深度学习
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一种可促进重组蛋白表达量和稳定性的多功能纯化标签的开发与利用 被引量:1
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作者 赵伟欣 刘松 +2 位作者 刘立明 陈坚 堵国成 《生物工程学报》 CAS CSCD 北大核心 2019年第4期626-635,共10页
自组装双亲短肽(Self-assembling amphipathic peptides,SAPs)是一类亲疏水氨基酸按一定规律分布、具有自聚合效应的短肽,融合在酶蛋白N端时,具有促进表达和稳定化的功能。根据前期研究结论,设计一条全新的基于SAPs (S1vw,HNANARARHNANA... 自组装双亲短肽(Self-assembling amphipathic peptides,SAPs)是一类亲疏水氨基酸按一定规律分布、具有自聚合效应的短肽,融合在酶蛋白N端时,具有促进表达和稳定化的功能。根据前期研究结论,设计一条全新的基于SAPs (S1vw,HNANARARHNANARARHNANARARHNARARAR)的可促进融合蛋白表达和稳定性,并可用于镍柱亲和层析的多功能短肽标签,在大肠杆菌表达系统中,将S1vw以PT-linker(PTPPTTPTPPTTPTP)融合在碱性果胶酶(Alkaline polygalacturonate lyase,PGL)、脂肪氧合酶(Lipoxygenase,LOX)及绿色荧光蛋白(Green fluorescent protein,GFP)的N末端时,与对应的野生型相比,PGL及LOX的粗酶活分别提高了3.1倍和1.89倍,GFP的荧光强度提高了16.22倍。S1vw的3种融合酶均可用镍柱进行亲和纯化,并具有较高的回收率。PGL及LOX在对应的热处理条件下,与野生型相比,半衰期分别提高了2.16倍和3.2倍。将GFP-S1vw在枯草芽孢杆菌及毕赤酵母表达系统中表达,发现在枯草芽孢杆菌中融合蛋白表达量提高明显,但在毕赤酵母中表达量几乎没有改变。说明在原核表达体系中,S1vw可作为一种新型的促表达、稳定化及可纯化的多功能标签。 展开更多
关键词 自组装双亲短肽 融合蛋白 表达量 热稳定性 纯化 表达宿主
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重组大肠杆菌全细胞催化L-苏氨酸合成2,5-二甲基吡嗪 被引量:1
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作者 于海波 徐建中 +1 位作者 刘立明 张伟国 《生物工程学报》 CAS CSCD 北大核心 2021年第1期228-241,共14页
2,5-二甲基吡嗪(2,5-dimethylpyrazine,2,5-DMP)在食品香料与医药方面具有重要的经济价值,工业上普遍采用环境不友好且反应条件苛刻的化学合成法来生产。文中结合代谢工程和辅因子工程策略设计高效催化L-苏氨酸合成2,5-DMP的全细胞催化... 2,5-二甲基吡嗪(2,5-dimethylpyrazine,2,5-DMP)在食品香料与医药方面具有重要的经济价值,工业上普遍采用环境不友好且反应条件苛刻的化学合成法来生产。文中结合代谢工程和辅因子工程策略设计高效催化L-苏氨酸合成2,5-DMP的全细胞催化剂,实现微生物转化法合成2,5-DMP。本研究首先分析了不同微生物来源的苏氨酸脱氢酶(Threonine dehydrogenase,TDH)对2,5-DMP合成的影响,发现来源于大肠杆菌Escherichia coli中EcTDH具有最佳的催化能力,2,5-DMP产量达到(438.3±23.7)mg/L。随后结合辅因子工程,通过引入乳脂链球菌Lactococcus cremoris中NADH氧化酶(NADH oxidase,LcNox E)并优化其表达方式发现通过融合表达EcTDH和Lc Nox E可平衡胞内NADH/NAD+水平,维持较高细胞存活率,进一步提高2,5-DMP产量。最后,通过阻断合成2,5-DMP的支路代谢途径,可以显著减少副产物积累,增加2,5-DMP产量,同时提高L-苏氨酸转化率。最终获得的重组菌EcΔkΔAΔBΔA/TDHEcNoxELc-PSst T在含有5 g/L L-苏氨酸的转化体系中于37℃、200 r/min孵化24 h,可积累(1095.7±81.3)mg/L的2,5-DMP,L-苏氨酸转化率达到76%,产物得率为0.288 g/(g L-苏氨酸)。因此,文中构建的重组菌可以实现高效催化L-苏氨酸合成2,5-DMP,具有一定的工业应用潜力。 展开更多
关键词 2 5-二甲基吡嗪 L-苏氨酸 全细胞催化 EcTDH-LcNoxE融合蛋白 L-苏氨酸转运蛋白SstT 辅因子工程
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Pharmacokinetics of monoclonal antibodies ]nd Fc-fusion proteins 被引量:13
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作者 liming liu 《Protein & Cell》 SCIE CAS CSCD 2018年第1期15-32,共18页
关键词 蛋白质 FC 抗体 熔化 MABS ND 相互作用能 再循环
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NADPH缺陷型Corynebacterium glutamicum重组菌株的构建及其性能分析
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作者 阮浩哲 刘立明 +1 位作者 张伟国 徐建中 《微生物学报》 CAS CSCD 北大核心 2021年第8期2442-2456,共15页
【目的】改造谷氨酸棒杆菌(Corynebacterium glutamicum)中NADPH合成途径,阻断胞内NADPH的合成,获得1株NADPH营养缺陷型菌株。【方法】通过失活L-赖氨酸高产菌C.glutamicum Lys-χ中葡萄糖-6-磷酸脱氢酶(Zwf)和苹果酸酶(MalE)并将NADP^... 【目的】改造谷氨酸棒杆菌(Corynebacterium glutamicum)中NADPH合成途径,阻断胞内NADPH的合成,获得1株NADPH营养缺陷型菌株。【方法】通过失活L-赖氨酸高产菌C.glutamicum Lys-χ中葡萄糖-6-磷酸脱氢酶(Zwf)和苹果酸酶(MalE)并将NADP^(+)依赖型异柠檬酸脱氢酶(NADP^(+)-Icdcg)替换成变形链球菌(Streptococcus mutans)中的NAD^(+)-Icdsm,阻断胞内NADPH的合成。随后结合辅因子工程,引入大肠杆菌(Escherichia coli)中膜结合吡啶核苷酸转氢酶(PntAB)并通过不同强度启动子控制PntAB的表达水平。最后,分析不同重组菌中胞内氧化还原水平和L-赖氨酸生产强度的变化。【结果】重组菌C.glutamicum Lys-χΔZMICg::ISm(即Lys-x1)胞内检测不到NADPH,为1株NADPH营养缺陷型菌株。该重组菌只在以葡萄糖酸为碳源的基础培养基中生长和积累L-赖氨酸,而以葡萄糖、丙酮酸、α-酮戊二酸和草酰乙酸为碳源时无法生长。此外,表达E.coli中的PntAB可回补重组菌Lys-χ1胞内NADPH的水平,但由于不同强度启动子控制PntAB表达水平不同,重组菌胞内NADPH水平也不同,并影响L-赖氨酸的生产强度。【结论】重组菌Lys-χ1可作为有效的底盘细胞,用于考察不同的NADPH再生策略,获得不同胞内NADPH水平的重组菌株,为进一步阐明NADPH调控微生物细胞生理代谢功能的机制提供研究基础。 展开更多
关键词 谷氨酸棒杆菌 NADPH缺陷体 NADPH再生 L-赖氨酸合成 氧化还原水平 启动子工程
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