Targeted expression, purification, and cleavage of fusion proteins from inclusion bodies in Escherichia coli

摘要

Therearemanyfusionproteinsystemsnowavailableforinsolubleexpression:TrpLE,ketosteroidisomerase,PurF,andPagP,forexample.Theidealfusionpartneriseffectiveatdirectingawidevarietyoftargetproteinsintoinclusionbodies,accumulatesinlargequantitiesinahighlypureform,andisreadilysolubilizedandpurifiedincommonlyuseddenaturants.Fusionpartnerremovalunderdenaturingconditionsisbiochemicallychallenging,requiringharshconditions(e.g.,cyanogenbromidein70%formicacid)thatcanresultinunwantedproteinmodifications.Recentadvancesinmetalion‐catalyzedpeptidebondcleavageallowformoremildconditions,andsomemethodsinvolvingnickelorpalladiumwilllikelysoonappearinmorebiologicalapplications...