Development of a minimal defined medium for recombinant human interleukin-3 production by Streptomyces lividans 66

• Published in: Biotechnology and bioengineering Vol. 99; no. 1; pp. 214 - 222
• Main Authors: Nowruzi, Keyvan, Elkamel, Ali, Scharer, Jeno M, Cossar, Doug, Moo-Young, Murray
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 2008; Wiley Subscription Services, Inc
• Subjects: Amino acids; Bacteria; Biochemistry; Cell Culture Techniques - methods; Cell Proliferation; Cell Survival; Culture Media - metabolism; Cytokines; distance-based multivariate analysis; Humans; Interleukin-3 - genetics; Interleukin-3 - metabolism; medium optimization; mixture experiments; Multivariate analysis; Protein Engineering - methods; recombinant human interleukin-3; Recombinant Proteins - metabolism; Streptomyces lividans; Streptomyces lividans - genetics; Streptomyces lividans - metabolism
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.21566

A systematic approach was developed to identify and optimize the essential amino acids in defined minimal medium for the production of recombinant human interleukin 3 (rHuIL-3) by Streptomyces lividans. Starvation trials were carried out initially to narrow down the number of probable essential amino acids from an initial number of 20 to 8. Then a screening mixture experiment was designed and performed with the eight identified amino acids and distance-based multivariate analysis was employed to rank the probable essential amino acids regarding both growth and product formation. Following this procedure, the search was narrowed to four amino acids (Asp, Leu, Met, and Phe). Finally, a mixture design experiment known as the simplex lattice design was carried out and the composition of the optimum minimal medium was found (Asp 53%, Met 5%, and Phe 42%). Biotechnol. Bioeng. 2008;99: 214-222. © 2007 Wiley Periodicals, Inc.