The cell now has a bunch of hydrogen ions (H⁺) trapped in the intermembrane space. They want to go back to the matrix, but they can’t just go anywhere — they have to go through a special protein channel called ATP synthase.

ATP Synthase is like a turbine:

As H⁺ ions flow through ATP synthase, their movement spins the turbine (like water turning a waterwheel). This spinning provides the energy to attach a phosphate group (P) to ADP (adenosine diphosphate), creating ATP (adenosine triphosphate).

NADH makes 3 ATPs
FADH₂ makes 2 ATPs

This whole process is called oxidative phosphorylation, and it’s the main way our body makes energy from food.

  1. NADH gives electrons to the first carrier.
  2. FADH₂ joins a little later in the chain.
  3. Electrons pass through carriers like Coenzyme Q (CoQ), Cytochrome b, Cytochrome c, and Cytochrome a.
  4. The last stop is oxygen, which accepts the electrons and forms water.

While most ATP is made in the electron transport chain, a few ATP molecules are made differently.

This method is called substrate-level phosphorylation. It happens during earlier steps like glycolysis and the Krebs cycle.

🔧 How it works:

An enzyme directly transfers a phosphate group from another molecule (called the substrate) to ADP, forming ATP.

📌 Example:

In glycolysis, an enzyme removes a phosphate from phosphoenolpyruvic acid (PEP) and gives it to ADP — making ATP and pyruvic acid.

✅ This method makes a small amount of ATP but is very fast and doesn’t need mitochondria or oxygen.

Let’s do the math:

  • Each NADH from the Krebs cycle = 3 ATP
  • Each FADH₂ = 2 ATP
  • NADH made during glycolysis must be moved into the mitochondria, and this uses 1 ATP per molecule, so each gives only 2 ATP.

Final Tally:

🔸 Glycolysis: 2 ATP (direct) + 2 NADH → 4 ATP (after cost)
🔸 Krebs Cycle: 2 ATP (direct) + 6 NADH → 18 ATP + 2 FADH₂ → 4 ATP
🔸 Total = 36 ATP

So, one glucose molecule can make up to 36 ATP if oxygen is present.