Improving the cost effectiveness of enhanced green fluorescent protein production using recombinant Escherichia coli BL21 (DE3): Decreasing the expression inducer concentration

Author(s)

Lopes, Camila ^[1] ; dos Santos, Nathalia Vieira ^[1] ; Dupont, Jana ^{[1, 2]} ; Pedrolli, Danielle Biscaro ^[1] ; Valentini, Sandro Roberto ^[3] ; Santos-Ebinuma, Valeria de Carvalho ^[1] ; Brandao Pereira, Jorge Fernando ^{[3, 1]}

Affiliation

^[1] Ao Paulo State Univ UNESP, Sch Pharmaceut Sci, Dept Bioproc & Biotechnol, Araraquara - Brazil

^[2] Univ Ghent, Fac Biosci Engn, Ghent - Belgium

^[3] Sao Paulo State Univ UNESP, Sch Pharmaceut Sci, Dept Biol Sci, Rodovia Araraquara Jau, Km 01 S-N, BR-14800903 Araraquara - Brazil

• Document type: Journal article
• Source: Biotechnology and Applied Biochemistry; v. 66, n. 4, p. 527-536, JUL 2019.
• Web of Science Citations: 0

Abstract

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-beta-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. (AU)

• FAPESP's process: 14/16424-7 - Optimization and scale-up of liquid-liquid extraction process with ionic liquids (ILs) as a sustainable tool for the separation of the anti-leukemia biopharmaceutical L-asparaginase (ASPase)
• Grantee: Jorge Pereira
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 18/50009-8 - Understanding the molecular interactions between ionic liquids and biopharmaceuticals: the key for a proper design of downstream processing
• Grantee: Jorge Pereira
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/19793-3 - Optimization and scale-up of novel Ionic-Liquid-based purification processes for recombinant green fluorescent protein produced by Escherichia coli "GFPurIL"
• Grantee: Sandro Roberto Valentini
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/07529-5 - Development of liquid-liquid purification process and incorporation in nanostructured polymeric matrix of recombinant green fluorescent protein produced by Escherichia coli
• Grantee: Nathalia Vieira Porphirio Veríssimo
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)