Dissolved oxygen concentration affects the accumulation of HIV-1 recombinant proteins in Escherichia coli.
A central problem in aerobic growth of any culture is the maintenance of dissolved oxygen concentration (DOC) above growth-limiting levels especially in high-cell density fermentations that are usually of the fed-batch type. Fermentor studies have been conducted to determine the influence of DOC on the production of heterologous proteins in Escherichia coli. The results demonstrated that there is a significant degree of product-to-product variation in the response of heterologous protein accumulation to DOC. For translational fusions of the human immunodeficiency virus-1 (HIV-1) proteins p24Gag and Env41, the imposition of a dissolved oxygen (DO) limitation resulted in 100 and 15% increases in the respective product yields. On the other hand, the imposition of a DO limitation had no effect on the production of a similar translational fusion of the HIV-1 protein p55Gag, and a large negative effect on the production of an influenza protein (C13). The stimulatory effects of DOC on p24Gag production were investigated further. The results of my studies suggested that the stimulatory effect observed at reduced agitation rates on p24Gag accumulation was owing to an oxygen effect and not a shear effect. Furthermore, the results of my investigations indicated that the effect a DOC had on the production of p24Gag was strongly influenced by the cell density at which the culture was induced.
Qoronfleh
M W
MW
SAIC Frederick, National Cancer Institute, Frederick Cancer Research and Development Center, MD 21702-1201, USA. ooronfle@ncifcrf.gov
D003001:Cloning, Molecular / Q000379:methods*; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D015686:Gene Products, env / Q000096:biosynthesis / Q000235:genetics; D016655:HIV Core Protein p24 / Q000096:biosynthesis* / Q000235:genetics; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis* / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D010100:Oxygen; D011994:Recombinant Proteins / Q000096:biosynthesis
D003470:Culture Media; D015686:Gene Products, env; D016655:HIV Core Protein p24; D015700:HIV Envelope Protein gp41; D011994:Recombinant Proteins; D010100:Oxygen