Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3)

An optimized cultivation condition is needed to maximize the functional green fluorescent protein (GFP) production. Six process variables (agitation rate, temperature, initial medium pH, concentration of inducer, time of induction, and inoculum density) were screened using the fractional factorial d...

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Main Authors: Chew, Few Ne, Tan, Siang Wen, Boo, Chern Huey, Tey, Ti Beng
Format: Article
Published: Taylor & Francis Group 2012
Subjects:
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author Chew, Few Ne
Tan, Siang Wen
Boo, Chern Huey
Tey, Ti Beng
author_facet Chew, Few Ne
Tan, Siang Wen
Boo, Chern Huey
Tey, Ti Beng
author_sort Chew, Few Ne
collection UMP
description An optimized cultivation condition is needed to maximize the functional green fluorescent protein (GFP) production. Six process variables (agitation rate, temperature, initial medium pH, concentration of inducer, time of induction, and inoculum density) were screened using the fractional factorial design. Three variables (agitation rate, temperature, and time of induction) exerted significant effects on functional GFP production in E. coli shake flask cultivation and were optimized subsequently using the Box-Behnken design. An agitation rate of 206 rpm at 31°C and induction of the protein expression when the cell density (OD(600nm)) reaches 1.04 could enhance the yield of functional GFP production from 0.025 g/L to 0.241 g/L, which is about ninefold higher than the unoptimized conditions. Unoptimized cultivation conditions resulted in protein aggregation and hence reduced the quantity of functional GFP. The model and regression equation based on the shake flask cultivation could be applied to a 2-L bioreactor for maximum functional GFP production.
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spelling UMPir93102018-05-14T06:41:59Z http://umpir.ump.edu.my/id/eprint/9310/ Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3) Chew, Few Ne Tan, Siang Wen Boo, Chern Huey Tey, Ti Beng TP Chemical technology An optimized cultivation condition is needed to maximize the functional green fluorescent protein (GFP) production. Six process variables (agitation rate, temperature, initial medium pH, concentration of inducer, time of induction, and inoculum density) were screened using the fractional factorial design. Three variables (agitation rate, temperature, and time of induction) exerted significant effects on functional GFP production in E. coli shake flask cultivation and were optimized subsequently using the Box-Behnken design. An agitation rate of 206 rpm at 31°C and induction of the protein expression when the cell density (OD(600nm)) reaches 1.04 could enhance the yield of functional GFP production from 0.025 g/L to 0.241 g/L, which is about ninefold higher than the unoptimized conditions. Unoptimized cultivation conditions resulted in protein aggregation and hence reduced the quantity of functional GFP. The model and regression equation based on the shake flask cultivation could be applied to a 2-L bioreactor for maximum functional GFP production. Taylor & Francis Group 2012 Article PeerReviewed Chew, Few Ne and Tan, Siang Wen and Boo, Chern Huey and Tey, Ti Beng (2012) Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3). Preparative Biochemistry and Biotechnology, 42 (6). pp. 535-550. ISSN 1082-6068. (Published) http://dx.doi.org/10.1080/10826068.2012.660903 DOI: Taylor & Francis Group, Taylor & Francis Group
spellingShingle TP Chemical technology
Chew, Few Ne
Tan, Siang Wen
Boo, Chern Huey
Tey, Ti Beng
Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3)
title Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3)
title_full Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3)
title_fullStr Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3)
title_full_unstemmed Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3)
title_short Statistical Optimization of Green Fluorescent Protein Production from Escherichia coli BL21(DE3)
title_sort statistical optimization of green fluorescent protein production from escherichia coli bl21 de3
topic TP Chemical technology
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AT tansiangwen statisticaloptimizationofgreenfluorescentproteinproductionfromescherichiacolibl21de3
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