Εξειδίκευση τύπου :	Άρθρο σε επιστημονικό περιοδικό
Τίτλος:	Escherichia coli strains with precise domain deletions in the ribonuclease RNase E can achieve greatly enhanced levels of membrane protein production
Δημιουργός/Συγγραφέας:	Vasilopoulou, Eleni Chroumpi, Tania [EL] Σκρέτας, Γιώργος[EN] Skretas, George
Ημερομηνία:	2024-02
Γλώσσα:	Αγγλικά
ISSN:	0961-8368 1469-896X
DOI:	10.1002/pro.4864
Άλλο:	38073126
Περίληψη:	Escherichia coli is one of the most widely utilized hosts for production of recombinant membrane proteins (MPs). Bacterial MP production, however, is usually accompanied by severe toxicity and low-level volumetric accumulation. In previous work, we had discovered that co-expression of RraA, an inhibitor of the RNA-degrading activity of RNase E, can efficiently suppress the cytotoxicity associated with the MP overexpression process and, simultaneously, enhance significantly the cellular accumulation of membrane-incorporated recombinant MPs in bacteria. Based on this, we constructed the specialized MP-producing E. coli strain SuptoxR, which can achieve dramatically enhanced volumetric yields of well-folded recombinant MPs. Ιn the present work, we have investigated whether domain deletions in the E. coli RNase E, which exhibit reduced ribonucleolytic activity, can result in suppressed MP-induced toxicity and enhanced recombinant MP production, in a manner resembling the conditions of rraA overexpression in E. coli SuptoxR. We have found that some strains encoding specific RNase E truncation variants can achieve significantly enhanced levels of recombinant MP production. Among these, we have found a single RNase E variant strain, which can efficiently suppress MP-induced toxicity and achieve greatly enhanced levels of recombinant MP production for proteins of both prokaryotic and eukaryotic origin. Based on its properties, and in analogy to the original SuptoxR strain, we have termed this strain SuptoxRNE22. E. coli SuptoxRNE22 can perform better than commercially available bacterial strains, which are frequently utilized for recombinant MP production. We anticipate that SuptoxRNE22 will become a widely utilized host for recombinant MP production in bacteria.
Τίτλος πηγής δημοσίευσης:	Protein science : a publication of the Protein Society
Τόμος/Κεφάλαιο:	33
Τεύχος:	2
Θεματική Κατηγορία:	[EL] Βιοτεχνολογία[EN] Biotechnology [EL] Βιοχημεία[EN] Biochemistry [EL] Μικροβιολογία[EN] Microbiology
Λέξεις-Κλειδιά:	Escherichia coli RNase E RraA SuptoxR Membrane protein Recombinant protein production
EU Grant:	HORIZON EUROPE Framework
EU Grant identifier:	819934 101079363 101087471
Κάτοχος πνευματικών δικαιωμάτων:	© 2023 The Authors.Protein Sciencepublished by Wiley Periodicals LLC on behalf of The Protein Society.
Όροι και προϋποθέσεις δικαιωμάτων:	This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
Ηλεκτρονική διεύθυνση στον εκδότη (link):	https://doi.org/10.1002/pro.4864
Εμφανίζεται στις συλλογές:	Ινστιτούτο Χημικής Βιολογίας - Επιστημονικό έργο