Εξειδίκευση τύπου :	Άρθρο σε επιστημονικό περιοδικό
Τίτλος:	SuptoxD2.0: A second-generation engineered Escherichia coli strain achieving further enhanced levels of recombinant membrane protein production
Δημιουργός/Συγγραφέας:	Michou, Myrsini Stergios, Angelos [EL] Σκρέτας, Γιώργος[EN] Skretas, George
Ημερομηνία:	2020-08
Γλώσσα:	Αγγλικά
ISSN:	0006-3592 1097-0290
DOI:	10.1002/bit.27378
Άλλο:	32383198
Περίληψη:	The bacterium Escherichia coli is among the most popular hosts for recombinant protein production, including that of membrane proteins (MPs). We have recently generated the specialized MP-producing E. coli strain SuptoxD, which upon co-expression of the effector gene djlA, is capable of alleviating two major bottlenecks in bacterial recombinant MP production: it suppresses the toxicity that frequently accompanies the MP-overexpression process and it markedly increases the cellular accumulation of membrane incorporated and properly folded recombinant MP. Combined, these two positive effects result in dramatically enhanced volumetric yields for various recombinant MPs of both prokaryotic and eukaryotic origin. Based on the observation that djlA is found in the genomes of various pathogenic bacteria, the aim of the present work was to investigate (a) whether other naturally occurring DjlA variants can exert the MP toxicity-suppressing and production-promoting effects similarly to the E. coli DjlA and (b) if we can identify a DjlA variant whose efficiency surpasses that of the E. coli DjlA of SuptoxD. We report that a quite surprisingly broad variety of homologous DjlA proteins exert beneficial effects on recombinant MP when overexpressed in E. coli. Furthermore, we demonstrate that the Salmonella enterica DjlA is an even more potent enhancer of MP productivity compared with the E. coli DjlA of SuptoxD. Based on this, we constructed a second-generation SuptoxD strain, termed SuptoxD2.0, whose MP-production capabilities surpass significantly those of the original SuptoxD, and we anticipate that SuptoxD2.0 will become a broadly utilized expression host for recombinant MP production in bacteria.
Τίτλος πηγής δημοσίευσης:	Biotechnology and bioengineering
Τόμος/Κεφάλαιο:	117
Τεύχος:	8
Σελίδες:	2434-2445
Θεματική Κατηγορία:	[EL] Βιοτεχνολογία[EN] Biotechnology [EL] Χημική Βιολογία[EN] Chemical Biology
Λέξεις-Κλειδιά:	DjlA Escherichia coli SuptoxD Recombinant membrane protein production Toxicity Escherichia coli Proteins HSP40 Heat-Shock Proteins Metabolic Engineering Escherichia coli Membrane Proteins Recombinant Proteins
Κάτοχος πνευματικών δικαιωμάτων:	Copyright © 2020 John Wiley & Sons, Inc. All rights reserved
Ηλεκτρονική διεύθυνση στον εκδότη (link):	https://onlinelibrary.wiley.com/doi/pdf/10.1002/bit.27378
Ηλεκτρονική διεύθυνση περιοδικού (link) :	https://onlinelibrary.wiley.com/journal/10970290
Εμφανίζεται στις συλλογές:	Ινστιτούτο Χημικής Βιολογίας - Επιστημονικό έργο