Department of Chemical Engineering, University of California, Berkeley, California 94720, USA.
Biotechnol Bioeng. 1989 Jan 20;33(4):477-86. doi: 10.1002/bit.260330413.
Glucose and glutamine are the main nutrients used by mammalian cells in culture. Each provides unique biosynthetic precursors but are complementary for production of other metabolites and energy. The transient and steady-state responses of hybridoma growth and metabolism to glucose pulse and step changes have been examined. Metabolic quotients are reported for oxygen, glucose, lactate, ammonia, glutamine, alanine, and other amino acids. The glucose consumption rate increased by 100-200% immediately after glucose was added to the reactor, and the increased glycolytic ATP production appears to be responsible for the concurrent rapid decrease in the oxygen consumption rate. The effects on glutamine consumption were delayed, probably due to buffering by the TCA cycle and interrelated pathways. A period of increased biosynthetic activity, as evidenced by an increase in the estimated specific ATP production rate and lower by-product yields from glutamine, preceded the increase in cell concentration after the glucose step change. The biosynthetic yield of cells from ATP was calculated, and it was estimated that maintenance accounted for about 60% of the energy used by the cells at a specific growth rate of 0.66 day(-1). The estimated 22% ATP production due to glycoysis was twice as great as that before the step change.
葡萄糖和谷氨酰胺是培养的哺乳动物细胞的主要营养物质。两者都提供独特的生物合成前体，但对于其他代谢物和能量的产生是互补的。已经研究了杂交瘤生长和代谢对葡萄糖脉冲和阶跃变化的瞬时和稳态响应。报告了氧、葡萄糖、乳酸盐、氨、谷氨酰胺、丙氨酸和其他氨基酸的代谢系数。葡萄糖添加到反应器后，葡萄糖的消耗率立即增加了 100-200％，增加的糖酵解 ATP 产生似乎是导致耗氧量同时快速下降的原因。对谷氨酰胺消耗的影响是延迟的，可能是由于 TCA 循环和相关途径的缓冲作用。在细胞浓度增加之前，存在一段时间的生物合成活性增加，这表现为估计的特定 ATP 产生率增加，以及来自谷氨酰胺的副产物产量降低。从 ATP 计算细胞的生物合成产率，并估计在特定生长速率为 0.66 天（-1）时，维持大约占细胞使用的能量的 60％。由于糖酵解而产生的估计 22％的 ATP 是阶跃变化之前的两倍。
