Εξειδίκευση τύπου :	Άρθρο σε επιστημονικό περιοδικό
Τίτλος:	Optimization of Recombinant Membrane Protein Production in the Engineered Escherichia coli Strains SuptoxD and SuptoxR
Δημιουργός/Συγγραφέας:	Michou M. Kapsalis C. Pliotas C. [EL] Σκρέτας, Γιώργος[EN] Skretas, George
Εκδότης:	American Chemical Society
Ημερομηνία:	2019
Γλώσσα:	Αγγλικά
ISSN:	2161-5063
DOI:	10.1021/acssynbio.9b00120
Άλλο:	PubMed ID: 31243979
Περίληψη:	Membrane proteins (MPs) execute a wide variety of critical biological functions in all living organisms and constitute approximately half of current targets for drug discovery. As in the case of soluble proteins, the bacterium Escherichia coli has served as a very popular overexpression host for biochemical/structural studies of membrane proteins as well. Bacterial recombinant membrane protein production, however, is typically hampered by poor cellular accumulation and severe toxicity for the host, which leads to low levels of final biomass and minute volumetric yields. In previous work, we generated the engineered E. coli strains SuptoxD and SuptoxR, which upon coexpression of the effector genes djlA or rraA, respectively, can suppress the cytotoxicity caused by MP overexpression and produce enhanced MP yields. Here, we systematically looked for gene overexpression and culturing conditions that maximize the accumulation of membrane-integrated and well-folded recombinant MPs in these strains. We have found that, under optimal conditions, SuptoxD and SuptoxR achieve greatly enhanced recombinant production for a variety of MP, irrespective of their archaeal, eubacterial, or eukaryotic origin. Furthermore, we demonstrate that the use of these engineered strains enables the production of well-folded recombinant MPs of high quality and at high yields, which are suitable for functional and structural studies. We anticipate that SuptoxD and SuptoxR will become broadly utilized expression hosts for recombinant MP production in bacteria.
Τίτλος πηγής δημοσίευσης:	ACS Synthetic Biology
Τόμος/Κεφάλαιο:	8
Τεύχος:	7
Σελίδες:	1631-1641
Θεματική Κατηγορία:	[EL] Βιολογία (Γενικά)[EN] Biology (General) [EL] Βιοτεχνολογία[EN] Biotechnology
Λέξεις-Κλειδιά:	DjlA E. coli SuptoxD E. coli SuptoxR recombinant membrane protein production RraA toxicity
Αξιολόγηση από ομότιμους (peer reviewed):	Ναι
Κάτοχος πνευματικών δικαιωμάτων:	© 2019 American Chemical Society.
Όροι και προϋποθέσεις δικαιωμάτων:	All Open Access, Green
Σημειώσεις:	MIS 5002636; Royal Society of Edinburgh, RSE; Carnegie Dunfermline Trust: OS000256; Tenovus: T15/41; University of St Andrews; European Commission, EC; State Scholarships Foundation, IKY; European Regional Development Fund, FEDER. This work was supported by a Greek State Scholarships Foundation (Idryma Kratikon Ypotrofion – IKY) scholarship, funded by the action “Strengthening human research potential through doctoral research” of the Partnership Agreement “Development of human potential, education and lifelong learning” 2014–2020, which is cofinanced by the European Structural and Investment Fund (ESIF) and the Greek State. We also acknowledge support by the project “Synthetic Biology: From omics technologies to genomic engineering (OMIC-ENGINE)” (MIS 5002636), which is implemented under the Action “Reinforcement of the Research and Innovation Infrastructure”, funded by the Operational Programme “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014-2020) and cofinanced by Greece and the European Union (European Regional Development Fund).
Εμφανίζεται στις συλλογές:	Ινστιτούτο Χημικής Βιολογίας - Επιστημονικό έργο