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Genetically Encoded Biosensor Engineering for Application in Directed Evolution
Mao Yin,Huang Chao,Zhou Xuan,Han Runhua,Deng Yu,Zhou Shenghu 한국미생물·생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.10
Although rational genetic engineering is nowadays the favored method for microbial strain improvement, building up mutant libraries based on directed evolution for improvement is still in many cases the better option. In this regard, the demand for precise and efficient screening methods for mutants with high performance has stimulated the development of biosensor-based highthroughput screening strategies. Genetically encoded biosensors provide powerful tools to couple the desired phenotype to a detectable signal, such as fluorescence and growth rate. Herein, we review recent advances in engineering several classes of biosensors and their applications in directed evolution. Furthermore, we compare and discuss the screening advantages and limitations of two-component biosensors, transcription-factor-based biosensors, and RNA-based biosensors. Engineering these biosensors has focused mainly on modifying the expression level or structure of the biosensor components to optimize the dynamic range, specificity, and detection range. Finally, the applications of biosensors in the evolution of proteins, metabolic pathways, and genome-scale metabolic networks are described. This review provides potential guidance in the design of biosensors and their applications in improving the bioproduction of microbial cell factories through directed evolution.
( Yanchen Yin ),( Youzhi Mao ),( Xiaolie Yin ),( Bei Gao ),( Dongzhi Wei ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.7
The filamentous fungus Aspergillus oryzae is a well-known expression host used to expresshomologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method.As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. ThePcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.
Mao, Yongle,Yang, Jiaqiang,Yin, Dejun,Chen, Yangsheng The Korean Institute of Power Electronics 2016 JOURNAL OF POWER ELECTRONICS Vol.16 No.6
Mechanical and electrical parameter uncertainties cause dynamic and static estimation errors of the rotor speed and position, resulting in performance deterioration of sensorless control systems. This paper applies an extended nonlinear observer to interior permanent magnet synchronous motors (IPMSM) for the simultaneous estimation of the rotor speed and position. Two compensation methods are proposed to improve the observer performance against parameter uncertainties: an on-line rotational inertia adjustment approach that employs the gradient descent algorithm to suppress dynamic estimation errors, and an equivalent flux error compensation approach to eliminate static estimation errors caused by inaccurate electrical parameters. The effectiveness of the proposed control strategy is demonstrated by experimental tests.
Synthesis method based on solution regions for planar four-bar straight-line linkages
Yin Lairong,Han Jianyou,Mao Cong,Huang Juan,Yang Tong 대한기계학회 2012 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.26 No.10
An analytical method for synthesizing and selecting desired four-bar straight-line mechanisms based on solution regions is presented. Given two fixed pivots, the point position and direction of the target straight-line, an infinite number of mechanism solutions can be produced by employing this method, both in the general case and all three special cases. Unifying the straight-line direction and the displacement from the given point to the instant center into the same form with different angles as parameters, infinite mechanism solutions can be expressed with different solution region charts. The mechanism property graphs have been computed to enable the designers to find out the involved mechanism information more intuitively and avoid aimlessness in selecting optimal mechanisms.
Yin, Yu-Yang,Liu, Yu-Jie,Zhu, Wei-Yun,Mao, Sheng-Yong Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.12
This study investigated the effects of acarbose addition on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro using batch cultures. Rumen fluid was collected from the rumens of three cannulated Holstein cattle fed forage ad libitum that was supplemented with 6 kg of concentrate. The batch cultures consisted of 8 mL of strained rumen fluid in 40 mL of an anaerobic buffer containing 0.49 g of corn grain, 0.21 g of soybean meal, 0.15 g of alfalfa and 0.15g of Leymus chinensis. Acarbose was added to incubation bottles to achieve final concentrations of 0.1, 0.2, and 0.4 mg/mL. After incubation for 24 h, the addition of acarbose linearly decreased (p<0.05) the total gas production and the concentrations of acetate, propionate, butyrate, total volatile fatty acids, lactate and lipopolysaccharide (LPS). It also linearly increased (p<0.05) the ratio of acetate to propionate, the concentrations of isovalerate, valerate and ammonia-nitrogen and the pH value compared with the control. Pyrosequencing of the 16S rRNA gene showed that the addition of acarbose decreased (p<0.05) the proportion of Firmicutes and Proteobacteria and increased (p<0.05) the percentage of Bacteroidetes, Fibrobacteres, and Synergistetes compared with the control. A principal coordinates analysis plot based on unweighted UniFrac values and molecular variance analysis revealed that the structure of the ruminal bacterial communities in the control was different to that of the ruminal microbiota in the acarbose group. In conclusion, acarbose addition can affect the composition of the ruminal microbial community and may be potentially useful for preventing the occurrence of ruminal acidosis and the accumulation of LPS in the rumen.
Molecular Epidemiology and Prevalence of Echovirus 30 in Zhejiang Province, China, from 2002 to 2015
( Yin Chen ),( Yi Sun ),( Juying Yan ),( Ziping Miao ),( Changping Xu ),( Yanjun Zhang ),( Haiyan Mao ),( Liming Gong ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.12
Echovirus serotype 30 (ECHO30) has been responsible for several recent worldwide outbreaks of viral meningitis. In Zhejiang Province, China, ECHO30 has been one of the main causes of viral meningitis for years. This study, using phylogenetic analysis of the VP1 gene, was performed to investigate the general molecular epidemiology and genetic patterns of ECHO30 circulating in Zhejiang Province between the years 2002 and 2015. The nucleotide sequences of ECHO30 VP1 showed that they were 64.8% identical with the prototype strain, Bastianni, while the amino acids were 84.9% identical. Phylogenetic analyses showed that ECHO30 in the Zhejiang area has diverged into two genotypes. Genotype I consists of strains isolated since 2002, whereas genotype II includes strains that were mainly isolated during the 2002 to 2004 outbreak. ECHO30 has been endemically circulating in both humans and the environment for a long period of time. Additionally, we evaluated the significance of recombination presented during the years 2005 to 2007 to demonstrate that recombination plays an important role in the prevalence of ECHO30 in the Zhejiang area.
Feasibility study of equal type, experienced and compound robots based on reliability theory
Zhe YIN,Yun-fei GUO,Mao-sheng LAI 제어로봇시스템학회 2011 제어로봇시스템학회 국제학술대회 논문집 Vol.2011 No.10
Through the introduction of the concept of reliability, events encountered by the robot are divided into three cases: the first is the one that the robot to can be sure to judge and finish the event, the second is that the robot can certainly not be able to determine if the events should be done. In both cases above the robot will be able to make a positive assessment; however, there is a fuzzy event, namely: the robot can not make sure, that this is the need to issue a request to the controller to get the new instruction, and then give a specific judge.
Yongle Mao,Jiaqiang Yang,Dejun Yin,Yangsheng Chen 전력전자학회 2016 JOURNAL OF POWER ELECTRONICS Vol.16 No.6
Mechanical and electrical parameter uncertainties cause dynamic and static estimation errors of the rotor speed and position, resulting in performance deterioration of sensorless control systems. This paper applies an extended nonlinear observer to interior permanent magnet synchronous motors (IPMSM) for the simultaneous estimation of the rotor speed and position. Two compensation methods are proposed to improve the observer performance against parameter uncertainties: an on-line rotational inertia adjustment approach that employs the gradient descent algorithm to suppress dynamic estimation errors, and an equivalent flux error compensation approach to eliminate static estimation errors caused by inaccurate electrical parameters. The effectiveness of the proposed control strategy is demonstrated by experimental tests.
Highly heat-resistant NF membrane modified by quinoxaline diamines for Li+ extraction from the brine
Huan-Yin Xie,Run-Hua Tang,Gui-E Chen,Zhen-Liang Xu,Haifang Mao 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.112 No.-
The Functional layers with excellent heat-resistence nanofiltration (HRN) property were designed by fixingthermal stabilized quinoxaline diamines (QHDA) on poly (m-phenyleneisophthalamide) (PMIA) substratethrough interfacial polymerization. QHDA was used as aqueous monomer while cinnamoylchloride (CNC), isophthaloyl chloride (IPC) and trimesoyl chloride (TMC) were served as organic monomersto realize the immobilization of thermally stable N heterocycles. The experimental results statedclearly that the compact package reaction of QHDA with TMC lead a raise of thermal stability and saltsrejection at high temperature stream. With the optimal TMC and 0.5 wt.% QHDA, the as-developed membraneachieved the excellent NF performance and thermal stability. The rejection of MgSO4 reached to94.6 % at 30 C and hardly decrease at 90 C. The reduction in rejection was satisfactory at high temperatures. The functional layer still showed highly stability after 10 h long-term operation at 90 C with only2.8 % reduction in MgSO4 rejection. Besides, the excellent Mg2+ and poor Li+ rejection made this membranehave huge potential in the application of extracting Li+ from brine. The separation factor ofMg2+-Li+ at high temperature was satisfactory. Over all, this study offered a prospect technology toexploit heat-resistant membranes for extraction lithium at high temperature.