2. Alcohol / Ethanol / Booze

3. Making Alcohol
The enzyme alcohol dehydrogenase plays a central role in the most ancient form of biotechnology: alcoholic fermentation.
Yeast and many bacteria produce alcohol dehydrogenases. These microbial enzymes catalyze the last step in the conversion of food into metabolic energy, creating ethanol.
Sugars are broken down and used for energy, forming ethanol as the waste product, which is excreted into the liquid surrounding the cell.
We have harnessed this process to produce alcoholic beverages: yeast is allowed to ferment grain sugars to form beer, and yeast is allowed to ferment grape juice to form wine.

4. Microbial ADH
Tetramer
4 x 352 amino acid residues
4 zinc ions (Zn++)
4 NAD cofactors

5. Making Alcohol
Alcohol dehydrogenases in microbes function as tetramers.
They are zinc-containing enzymes that utilize glucose.
Each glucose molecule is broken down in a 10-step process called glycolysis. The product of glycolysis is two three-carbon sugars, called pyruvates, and ATP (adenosine triphosphate).
The two pyruvates are then converted into ethanol and carbon dioxide.
The overall process of fermentation is to convert glucose sugar to alcohol and carbon dioxide gas:
C6H12O6        2 CH3CH2OH               2 CO2    sugar     alcohol             carbon dioxide gas (glucose)    (ethyl alcohol or ethanol)

6. Making Alcohol

7. Making Alcohol

8. Breaking Down Alcohol
In 1997, Americans drank an average of 2 gallons (7.6 liters) of alcohol per person. This translates roughly into one six-pack of beer, two glasses of wine and three or four mixed drinks per wee.
So while recovering from the excesses at the Anthill Pub [last night] after it reopens next fall, we might ponder the human alcohol dehydrogenase enzyme, which ceaselessly battles all the beer & wine that we have consumed.

9. Breaking Down Alcohol
Alcohol dehydrogenase is our primary defense against alcohol, a toxic molecule that compromises the function of our nervous system.
The high levels of alcohol dehydrogenase in our liver and stomach detoxify about one drink each hour.
The alcohol is converted to acetaldehyde, an even more toxic molecule and the main cause of hangovers!
Acetaldehyde in turn is converted to acetate and other molecules that are easily processed by our cells.

10. Human ADH
Homodimer (two molecules)
2 x 373 amino acid residues
6 zinc ions (Zn++)
2 NAD cofactors

11. Human ADH Microbial

12. Breaking Down Alcohol CH3CH2OH + 2 NAD  CH3CHO + 2 NADH
Alcohol dehydrogenase
CH3CH2OH    + 2 NAD    CH3CHO    + 2 NADH
alcohol          cofactor  aldehyde cofactor (ethanol) (acetaldehyde)
Acetaldehyde dehydrogenase 2
CH3CHO H2O    CH3COOH
  aldehyde acid
(acetaldehyde) (acetic acid or vinegar)

13. Breaking Down Alcohol CH3CHO + H2O  CH3COOH
Acetaldehyde dehydrogenase 2
CH3CHO H2O    CH3COOH
  aldehyde acid
(acetaldehyde) (acetic acid or vinegar)
The acetic acid can be used to form fatty acids (watch that waistline!), or it can be further broken down into CO2 and water.

14. Dangers of Alcohol
Alcohol dehydrogenase provides a line of defense against a common toxin in our environment.
But alcohol dehydrogenase also modifies other alcohols, sometimes producing even more dangerous products:
Methanol, which is commonly used to “denature” ethanol rendering it undrinkable, is converted to formaldehyde by alcohol dehydrogenase.
The formaldehyde then causes severe damage, attacking proteins and embalming them.
Small amounts of methanol cause blindness, as the sensitive proteins in the retina are attacked, and larger amounts, perhaps a glassful, lead to widespread damage and death.

15. Breaking Down Methanol
Alcohol dehydrogenase
CH3CH2OH    + 2 NAD    CH3CHO    + 2 NADH
alcohol          cofactor  aldehyde cofactor (ethanol) (acetaldehyde)
Alcohol dehydrogenase
CH3OH      NAD    CH3CHO    + 2 NADH
alcohol    cofactor  aldehyde cofactor (methanol) (formaldehyde)

16. Structure (Form) & Function
Our bodies create at least nine different forms of alcohol dehydrogenase, each with slightly different properties.
Most of these are found primarily in the liver, including the b3 form
The s form is found in the lining of the stomach.
Each enzyme is composed of two subunits.
Ethanol is not the only target or substrate of these enzymes, they also make important modifications to retinol, steroids, and fatty acids.

18. Structure (Form) & Function
Human alcohol dehydrogenases use two “helpers” to perform their reaction on ethanol.
The first are zinc ions (Zn++), which are used to hold and position the alcohol group on ethanol.
The second is the NAD cofactor (constructed using the vitamin niacin), which actually performs the chemical reaction.
The zinc atom, shown in light blue, is cradled by three amino acids from the protein: cysteine 46 to the left, cysteine 174 to the right, and histidine 67 above. The ethanol, shown in green and magenta, binds to the zinc and is positioned next to the NAD cofactor, which extends below the ethanol molecule in this illustration.