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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| Format: | Article |
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Taylor & Francis Group
2012
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| Online Access: | http://umpir.ump.edu.my/id/eprint/9310/ |
| _version_ | 1848818459312390144 |
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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 |
| building | UMP Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T01:38:00Z |
| format | Article |
| id | ump-9310 |
| institution | Universiti Malaysia Pahang |
| institution_category | Local University |
| last_indexed | 2025-11-15T01:38:00Z |
| publishDate | 2012 |
| publisher | Taylor & Francis Group |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | ump-93102018-05-14T06:41:59Z 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 |
| url | http://umpir.ump.edu.my/id/eprint/9310/ http://umpir.ump.edu.my/id/eprint/9310/ http://umpir.ump.edu.my/id/eprint/9310/ |