Final answer:
The substrate used in the last step of glycolysis is pyruvate, which is converted from phosphoenolpyruvate (PEP) by the enzyme pyruvate kinase. This step produces ATP through substrate-level phosphorylation.
Step-by-step explanation:
The substrate used in the last step of glycolysis is pyruvate. In the final step of glycolysis, the enzyme pyruvate kinase catalyzes the conversion of phosphoenolpyruvate (PEP) to pyruvate. This step produces a second molecule of ATP by substrate-level phosphorylation, along with pyruvic acid or pyruvate, which can be further catabolized or converted into other molecules.