As glucose is introduced into the glycolytic pathway, it is first phosphorylated to create glucose-6-phosphate. That will then be converted to fructose-6-phosphate via phosphoglucose isomerase. That product will then be phosphorylated once more via phosphofructokinase-1 to create fructose-1,6-bisphosphate. Glucose-1,6-bisphosphate is never an observed intermediate in glycolysis.

Cellular respiration is a long process, and so it is easiest to break it into the following steps:

Step 1: Glycolysis

Step 2: Pyruvate decarboxylation

Step 3: Krebs cycle

Step 4: Oxidative phosphorylation

In the above steps, ATP is only produced by substrate-level phosphorylation in glycolysis and during the Krebs cycle.

In glycolysis, two molecules of pyruvate are produced for every molecule of glucose oxidized. During this process, two ATP molecules are consumed, but four are produced via substrate-level phosphorylation.

In the Krebs cycle, each pass of pyruvate through the cycle generates one molecule of GTP, which is subsequently used to generate a molecule of ATP via substrate-level phosphorylation. Thus, one molecule of ATP is produced via substrate-level phosphorylation per molecule of pyruvate oxidized. But remember that glycolysis produces two molecules of pyruvate for each molecule of glucose oxidized. Hence, the Krebs cycle will contribute a total of two molecules of ATP per glucose molecule oxidized.

Since we have a total of four moles ATP from glycolysis and two moles of ATP from the Krebs cycle (one per pyruvate), we have a cumulative production of six moles of ATP generated by substrate-level phosphorylation per mole of glucose oxidized.

The first and third steps of glycolysis involve energy consumption in the form of ATP. A phosphate group is added to glucose, and fructose-6-phosphate. In the seventh and tenth steps of glycolysis, ADP is phosphorylated at the level of the substrate into ATP. Since this is after glucose had been split into two three-carbon molecules, each molecule of glucose results in four ATP produced. However, since two were consumed early in glycolysis, the net ATP production is 2.

The first step of glycolysis is the addition of a phosphate group to glucose to form glucose-6-phosphate. The third step of glycolysis is the addition of another phosphate group to fructose-6-phosphate to form fructose-1,6-bisphosphate. The conversion of ATP to ADP is needed to supply the phosphate group in both of these reactions. These are the only two reactions in glycolysis where ATP is used to to add phosphate groups.

Glycolysis produces 4 ATP molecules. However, 2 ATP molecules are required to initiate glycolysis. Subtracting these two numbers gives the net ATP yield from glycolysis--2 ATP molecules.

Although glycolysis will ultimately produce 4 ATP, there is an initial requirement of 2 ATP for it to begin. The conversion of glucose to glucose-6-phosphate and the conversion of fructose-6-phosphate to fructose-1,6-bisphosphate both require ATP.  

The first and third steps of glycolysis are both energetically unfavorable. This means they will require an input of energy in order to continue forward. Per glucose molecule, 1 ATP is required for each of these steps. Therefore, a total of 2 ATP is needed during the energy investment phase of glycolysis.  

In the glycolytic pathway, 2 molecules of ATP must be used. The purpose of these molecules is to phosphorylate 2 intermediates in the pathway:

1. Glucose must be phosphorylated to glucose-6-phosphate.

2. Fructose-6-phosphate must be phosphorylated to fructose-1,6-bisphosphate.

Hexokinase and glucokinase are two enzymes that serve similar roles but have different characteristics. Hexokinase is found in all tissues, is inhibited by glucose 6 phosphate, and is not induced by insulin. It has a physiologic role of providing cells with a basal level of glucose 6 phosphate necessary for energy production.

The reaction turning glyceraldehyde-3-phosphate into 1,3-bisphosphoglycerate is shown below

This step of glycolysis does not hydrolyze or generate ATP, even though a phosphate group was added onto the glyceraldehyde-3-phosphate. The energy released when  is reduced to , sometimes referred to as the energy of oxidation (of glyceraldehyde-3-phosphate).