The mechanistic target of rapamycin (mTOR) protein kinase exists in 2 different complexes. Genetic inhibition of mTORC2 in the liver disrupts whole body glucose homeostasis. The objective of this study was to determine the mechanism of mTORC2 regulation of hepatic glucose metabolism. We performed quantitative mass spectrometry analysis of cultivated primary hepatocytes from 9-wk-old C57BL/6 wild type mice or mice which Rictor, an essential protein of mTORC2, had been deleted exclusively in the liver. Using 5% false discovery rate, we detected 1703 proteins significantly affected by Rictor deletion. As previously described, we found that gluconeogenic enzymes were upregulated, while proteins involved in fatty acid synthesis were downregulated in Rictor knock out (RKO) hepatocytes. Additionally, we found that rate limiting glycolytic enzymes were downregulated, while enzymes involved in TCA cycle, mitochondrial fatty acid transport and oxidation, ketone bodies and cholesterol synthesis were upregulated in RKO hepatocytes. Wild type and RKO hepatocytes were cultivated and 24 h later oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), as a mean of glycolytic function, were measured in vitro. While OCR was not affected by Rictor deletion, ECAR and ECAR/OCR were significantly reduced, suggesting that RKO hepatocytes reprogram energy sources to sustain respiration rate. We did not detect changes in OCR in the presence of a mitochondrial fatty acid transport inhibitor. Therefore, we measured plasma levels of BHBA and cholesterol, 2 other potential metabolic fates of acetyl CoA, in addition to TCA cycle, derived from fatty acid oxidation. While plasma free cholesterol and cholesterol esters were not affected by hepatic Rictor deletion, fasting plasma BHBA was significantly increased. Our results suggest that to sustain energy levels under dysregulated gluconeogenesis, RKO hepatocytes increase fatty acid oxidation and ketone body production.