Viruses alter the metabolism of host cells for their own replicative advantage. This study aimed to determine if the SARS-CoV-2 Spike glycoprotein plays a role in this process by interacting with, and exerting a direct effect on, enzymes of central carbon metabolism, specifically human Lactate Dehydrogenase (HsLDH) and Malate Dehydrogenase (HsMDH). Fusions of the Spike protein, HsLDH and HsMDH to the C-terminal half and N-terminal half of ubiquitin were used to test for in vivo interactions in yeast using the Split-Ubiquitin system. Using proteins expressed in yeast and bacterial systems, the effect exerted by Spike could be determined in vitro through enzyme activity assays. The split-ubiquitin system detected the in vivo interaction of Spike with HsLDH and HsMDH, as well as the interactions between Spike and all three isoenzymes of Saccharomyces cerevisiae malate dehydrogenase (ScMDH). Spike could only be expressed in a yeast system, and exposure to the viral protein had no effect on HsLDH activity in vitro. However, exposure to Spike increased ScMDH activity. Therefore, this report provides some of the first evidence for Spike’s involvement in the metabolic reprogramming of HsMDH. Such new knowledge regarding the metabolic targets of Spike is useful in the discovery of therapeutic targets.