Εξειδίκευση τύπου :	Άρθρο σε επιστημονικό περιοδικό
Τίτλος:	Second-Generation Escherichia coli SuptoxR Strains for High-Level Recombinant Membrane Protein Production
Δημιουργός/Συγγραφέας:	Vasilopoulou, Eleni Giannakopoulou, Artemis Kapsalis, Charalampos Michou, Myrsini Michoglou-Sergiou, Aristeidis [EL] Κολίσης, Φραγκίσκος Ν.[EN] Kolisis, Fragiskos N.‏ [EL] Σκρέτας, Γιώργος[EN] Skretas, George
Εκδότης:	American Chemical Society
Ημερομηνία:	2022-08-19
Γλώσσα:	Αγγλικά
ISSN:	2161-5063 2161-5063
DOI:	10.1021/acssynbio.1c00598
Άλλο:	35922033
Περίληψη:	Escherichia coli is one of the most widely utilized hosts for recombinant protein production, including that of membrane proteins (MPs). We have recently engineered a specialized E. coli strain for enhanced recombinant MP production, termed SuptoxR. By appropriately co-expressing the effector gene rraA, SuptoxR can suppress the high toxicity, which is frequently observed during the MP-overexpression process, and, at the same time, enhance significantly the cellular accumulation of membrane-incorporated and properly folded recombinant MP. The combination of these two beneficial effects results in dramatically enhanced volumetric yields for various prokaryotic and eukaryotic MPs. Here, we engineered second-generation SuptoxR strains with further improved properties, so that they can achieve even higher levels of recombinant MP production. We searched for naturally occurring RraA variants with similar or improved MP toxicity-suppressing and production-promoting effects to that of the native E. coli RraA of the original SuptoxR strain. We found that the RraA proteins from Proteus mirabilis and Providencia stuartii can be even more potent enhancers of MP productivity than the E. coli RraA. By exploiting these two newly identified RraAs, we constructed two second-generation SuptoxR strains, termed SuptoxR2.1 and SuptoxR2.2, whose MP-production capabilities often surpass those of the original SuptoxR significantly. SuptoxR2.1 and SuptoxR2.2 are expected to become widely useful expression hosts for recombinant MP production in bacteria.
Τίτλος πηγής δημοσίευσης:	ACS synthetic biology
Τόμος/Κεφάλαιο:	11
Τεύχος:	8
Σελίδες:	2599 - 2609
Θεματική Κατηγορία:	[EL] Βιοτεχνολογία[EN] Biotechnology [EL] Χημική Βιολογία[EN] Chemical Biology [EL] Γενετική[EN] Genetics [EL] Μικροβιολογία[EN] Microbiology
Λέξεις-Κλειδιά:	Escherichia coli RraA SuptoxR Membrane protein Recombinant production Toxicity
EU Grant:	HFRI PhD Fellowship Development of second-generation specialized bacterial strains for high-level recombinant membrane protein production ProMiDis Synthetic Biology: From omics technologies to genomic engineering (OMIC-ENGINE)
EU Grant identifier:	Fellowship Number: 961 MIS code 5050546 819934 MIS 5002636
Κάτοχος πνευματικών δικαιωμάτων:	© 2022 The Authors
Ηλεκτρονική διεύθυνση στον εκδότη (link):	https://pubs.acs.org/doi/10.1021/acssynbio.1c00598
Σημειώσεις:	The Supporting Information is available at: https://pubs.acs.org/doi/10.1021/acssynbio.1c00598
Εμφανίζεται στις συλλογές:	Ινστιτούτο Χημικής Βιολογίας - Επιστημονικό έργο