Statistical medium formulation and process modeling by mixture design of experiment for peptide overexpression in recombinant Escherichia coli.
The medium formulation and robust process modeling for anti-HIV peptide (T-20) production by recombinant Escherichia coli overexpression were studied by employing a crossed experimental design. The crossed design, a mixture design combined with process factor (induction duration), was used to find the optimal medium formulation and process time. The optimal settings for three major components (7.75 mL of NPK sources, 5.5 mL of glucose, and 11.75 mL of MgSO4) characterized by %T-20 (14.45%), the proportion of peptide to the total protein, were observed in a total of 100 mL of medium inducted at an optical density of 0.67 with 0.7 mM isopropyl-beta-D-thiogalactopyranoside) for a 3-h induction duration at shake-flask scale. These conditions were further investigated to find robust process conditions (8.2 mL of NPK sources, 5.6 mL of glucose, and 11.3 mL of MgSO4, and a 3.5-h induction duration time) for T-20 production (13.9%) by applying propagation of error.
Lee
Kwang-Min
KM
Cellular Metabolic Engineering Lab., Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong Yuseong-gu Daejeon, 305-701, South Korea. Klee0922@yahoo.co.kr
Rhee
Chang-Hoon
CH
Kang
Choong-Kyung
CK
Kim
Jung-Hoe
JH
D019380:Anti-HIV Agents / Q000737:chemistry; D001709:Biotechnology / Q000379:methods*; D002626:Chemistry, Pharmaceutical / Q000379:methods; D003470:Culture Media; D015195:Drug Design; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015233:Models, Statistical; D010455:Peptides / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins; D012044:Regression Analysis; D013997:Time Factors
D019380:Anti-HIV Agents; D003470:Culture Media; D010455:Peptides; D011994:Recombinant Proteins