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Under these conditions, pyruvate, an intermediate compound produced during the metabolism of glucose, is converted to lactate by lactate dehydrogenase rather than entering into the tricarboxylic acid cycle, where it is further oxidized through the activity of pyruvate carboxylase (Fig. 3.44).
To counteract the acidification of the me dium from lactate secretion, the human pyruvate carboxylase gene was cloned into an expression vector under the control of the cytomegalovirus (CMV) promoter and the SV40 polyadenylation signals and transfected into
When the pyruvate carboxylase gene was stably integrated into the CHO genome and expressed, the enzyme was detected in the mi tochondria, where glucose is degraded.
Figure 3.44 When oxygen is present, pyruvate, which is formed from glu cose during glycolysis, is converted by the enzyme pyruvate carboxylase to an intermediate compound in the tricar boxylic acid (TCA) cycle.
However, under lowoxygen conditions, such as those found in large bioreactors, pyruvate carboxylase has a low level of activity.
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