With worldwide attention on renewable energy and climate change,metabolic engineering of the fatty acid biosynthetic pathway has become an active area of research,with a view to enhance production of biofuels.Indeed,t...With worldwide attention on renewable energy and climate change,metabolic engineering of the fatty acid biosynthetic pathway has become an active area of research,with a view to enhance production of biofuels.Indeed,this pathway has already been extensively studied in Escherichia coli.Nevertheless,little is known about the absolute abundance of the enzymes involved,information that may be valuable for engineering,such as the optimal molar ratios of different proteins.In this study,we use protein standard absolute quantification(PSAQ)to measure the absolute abundance of proteins that catalyze fatty acid biosynthesis in E.coli.In addition,the changes of protein abundance were analyzed by comparing the differences between high-yield and the background strain.Our work highlights opportunities to enhance fatty acid production by measuring protein molar ratios and identifying catalytic and regulatory bottlenecks.More importantly,our results provide evidence that PSAQ is a generally valuable tool to investigate metabolic pathways.展开更多
基金grants from the 863(2012AA02A701)973(2011CBA00800,2012CB721000)Project from the Ministry of Science and Technology of the People’s Republic of China and the National Natural Science Foundation of China(31170096,31222002)as well as the project from Key Laboratory of Biofuels,Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences(CASKLB201301)Science and Technology Department of Hubei Province and J1 Biotech Co.Ltd.(2014091610010595).
文摘With worldwide attention on renewable energy and climate change,metabolic engineering of the fatty acid biosynthetic pathway has become an active area of research,with a view to enhance production of biofuels.Indeed,this pathway has already been extensively studied in Escherichia coli.Nevertheless,little is known about the absolute abundance of the enzymes involved,information that may be valuable for engineering,such as the optimal molar ratios of different proteins.In this study,we use protein standard absolute quantification(PSAQ)to measure the absolute abundance of proteins that catalyze fatty acid biosynthesis in E.coli.In addition,the changes of protein abundance were analyzed by comparing the differences between high-yield and the background strain.Our work highlights opportunities to enhance fatty acid production by measuring protein molar ratios and identifying catalytic and regulatory bottlenecks.More importantly,our results provide evidence that PSAQ is a generally valuable tool to investigate metabolic pathways.
