Improving the cost effectiveness of enhanced green fluorescent protein production using recombinant Escherichia coli BL21 (DE3): Decreasing the expression inducer concentration
Autor: | Nathalia Vieira dos Santos, Jana Dupont, Valéria de Carvalho Santos-Ebinuma, Camila Lopes, Danielle Biscaro Pedrolli, Sandro Roberto Valentini, Jorge F. B. Pereira |
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Rok vydání: | 2019 |
Předmět: |
0106 biological sciences
Cost effectiveness Globular protein Green Fluorescent Proteins Biomedical Engineering lac operon Bioengineering Enhanced green fluorescent protein medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology Green fluorescent protein 03 medical and health sciences 010608 biotechnology Drug Discovery medicine Escherichia coli Inducer 030304 developmental biology chemistry.chemical_classification 0303 health sciences Recombinant escherichia coli Process Chemistry and Technology General Medicine Recombinant Proteins Cell biology chemistry Molecular Medicine Biotechnology |
Zdroj: | Biotechnology and applied biochemistry5. References. 66(4) |
ISSN: | 1470-8744 |
Popis: | Green fluorescent protein (GFP) is a globular protein used as biosensor and biomarker in medical and industrial fields. However, due to the expensive production costs of expressing proteins using high-cost inducers like isopropyl-β-d-1-thiogalactopyranoside (IPTG), the number of GFP applications are still scarce. This work studied the production of enhanced GFP (EGFP) using Escherichia coli BL21 (DE3) [pLysS; pET28(a)], aiming to increase its yield and reduce costs. First, the influence of agitation rate, induction time, and concentration of IPTG in the production of EGFP was evaluated, but only the first two parameters were significant. Subsequently, aiming to reduce costs related to the use of inducer, the IPTG concentration (0.005, 0.010, and 0.025 mM) was decreased and, interestingly, the production levels were maintained or increased. These results show that a proper choice of production conditions, particularly through the decrease of inducer concentration, is effective to reduce the upstream production costs and guarantee high EGFP expression. |
Databáze: | OpenAIRE |
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