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  • Title: Glutamine-based dipeptides are utilized in mammalian cell culture by extracellular hydrolysis catalyzed by a specific peptidase.
    Author: Christie A, Butler M.
    Journal: J Biotechnol; 1994 Nov 15; 37(3):277-90. PubMed ID: 7765576.
    Abstract:
    A murine hybridoma (CC9C10) was grown in media containing alanyl-glutamine (ala-gln) or glycyl-glutamine (gly-gln) as a substitute for glutamine. High cell yields were obtained in the presence of 6 mM ala-gln or 20 mM gly-gln. The final cell yield in gly-gln was 14% higher than in gln. Monoclonal antibody productivity was comparable in gln, ala-gln or gly-gln. Substrate utilisation and metabolism was affected by the presence of the dipeptides, particularly with gly-gln. The specific consumption rates of glucose and six amino acids were reduced. Also the accumulation of ammonia and lactate was significantly lower. The higher concentration of gly-gln was necessary for cell growth because of the presence of a peptidase with a lower affinity for gly-gln. The peptidase activity of the basal medium and serum was low. However, a substantial peptidase activity was found in the cytosolic fraction of the cells with an apparent Km of 1.2 mM with respect to ala-gln and 14 mM with respect to gly-gln. It is proposed that the mechanism of dipeptide utilisation involves extracellular hydrolysis by this peptidase following its release into the culture medium.
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