Step 1: Understanding ATP investment in glycolysis.
The glycolytic pathway begins with glucose being phosphorylated to form glucose-6-phosphate. This process requires energy in the form of ATP. A second ATP molecule is used to phosphorylate fructose-6-phosphate into fructose-1,6-bisphosphate.
Step 2: ATP molecules used.
Two ATP molecules are consumed during the energy investment phase of glycolysis:
One ATP is used in the conversion of glucose to glucose-6-phosphate.
One ATP is used in the conversion of fructose-6-phosphate to fructose-1,6-bisphosphate.
\[
\therefore \text{The correct answer is: 2.}
\]