Alcohol dehydrogenase: Wikis


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alcohol dehydrogenase
Protein ADH5 PDB 1m6h.png
Crystallographic structure of the
homodimer of human ADH5.[1]
EC number
CAS number 9031-72-5
IntEnz IntEnz view
ExPASy NiceZyme view
MetaCyc metabolic pathway
PRIAM profile
PDB structures
Gene Ontology AmiGO / EGO

Alcohol dehydrogenases (ADH) (EC are a group of seven dehydrogenase enzymes that occur in many organisms and facilitate the interconversion between alcohols and aldehydes or ketones with the reduction of NAD+ to NADH. In humans and many other animals, they serve to break down alcohols which could otherwise be toxic; in yeast and many bacteria, some alcohol dehydrogenases catalyze the opposite reaction as part of fermentation.



Genetic evidence from comparisons of multiple organisms, showed that a glutathione dependent formaldehyde dehydrogenase, identical to an ADH3, probably is the ancestral enzyme for the entire ADH family.[2][3]. Early on in evolution, an effective method for eliminating endogenous and exogenous formaldehyde was important and this capacity has conserved the ancestral ADH3 through time. From genetic duplications of ADH3, followed by series of mutations, the other ADHs evolved.[2][3] The ability to produce ethanol from sugar is believed to have evolved in yeast. This feature is not rational from an energetic point of view, but by making alcohol in such high concentrations so that they were toxic to other organisms, yeast cells could effectively eliminate their competition. Since rotting fruit can contain more than 4% of ethanol, animals eating the fruit needed a system to metabolize exogenous ethanol. This can explain the need for an ethanol active ADH in other species than yeast.


The first ever isolated alcohol dehydrogenase (ADH) was purified in 1937 from Saccharomyces cerevisiae (baker’s yeast).[4] Many aspects of the catalytic mechanism for the horse liver ADH enzyme was investigated by Hugo Theorell and coworkers.[5] ADH was also one of the first oligomeric enzymes that got its amino acid sequence and three dimensional structure determined.[6][7][8]

In the beginning of 1960 it was discovered in fruit flies of the genus Drosophila.[9]


The alcohol dehydrogenases comprise a group of several isozymes that facilitate the conversion of toxic alcohols to aldehydes. In mammals this is a redox (reduction/oxidation) reaction involving the coenzyme nicotinamide adenine dinucleotide (NAD+)

Alcohol dehydrogenase is a dimer with a mass of 80 kDa.[10]

Alcohol dehydrogenase is responsible for catalyzing oxidation of primary and secondary alcohols to aldehydes and ketones, respectively, and also can affect the reverse reaction. It does not work well with primary alcohols. Instead, it works the best with secondary and cyclic alcohols.[9]

Oxidation of alcohol

Mechanism of action in humans


  1. Binding of the coenzyme NAD+;
  2. Binding of the alcohol substrate by coordination to zinc;
  3. Deprotonation of His-51;
  4. Deprotonation of nicotinamide ribose;
  5. Deprotonation of Ser-48;
  6. Deprotonation of the alcohol;
  7. Hydride transfer from the alkoxide ion to NAD+, leading to NADH and a zinc bound aldehyde or ketone;
  8. Release of the product aldehyde;

The mechanism in yeast and bacteria is the reverse of this reaction. These steps are supported through kinetic studies.[10]

Involved subunits

The substrate is coordinated to the zinc and this enzyme has two zinc atoms per subunit. One is the active site, which is involved in catalysis. In the active site, the ligands are Cys-46, Cys-174,His-67 and one water molecule. The other subunit is involved with structure. In this mechanism, the hydride from the alcohol goes to NAD+. Crystal structures indicate that the His-51 deprotonates the nicotinamide ribose, which deprotonates Ser-48. Finally, Ser-48 deprotonates the alcohol, making it an aldehyde.[10] From a mechanistic perspective, if the enzyme adds hydride to the re face of NAD+, the resulting hydrogen is incorporated into the pro-R position. Enzymes that add hydride to the re face are deemed Class A dehydrogenases.

Active site

The active site of alcohol dehydrogenase

The active site consists of a zinc atom, His-67, Cys-174, Cys-46, Ser-48, His-51, Ile-269, Val-292, Ala-317, and Phe-319. The zinc coordinates the substrate(alcohol). The zinc is coordinated by Cys-146, Cys-174, and His-67. Phe-319, Ala-317, His-51, Ile-269 and Val-292 stabilize NAD+ by forming hydrogen bonds. His-51 and Ile-269 form hydrogen bonds with the alcohols on nicotinamide ribose. Phe-319, Ala-317 and Val-292 form hydrogen bonds with the amide on NAD+.[10]

Structural zinc site

The structural zinc binding motif in alcohol dehydrogenase from a MD simulation

Mammalian alcohol dehydrogenases also has a structural zinc site. This Zn ion plays a structural role and is crucial for protein stability. The structures of the catalytic and structural zinc sites in horse liver alcohol dehydrogenase (HLADH) as revealed in crystallographic structures which has been studied computationally with quantum chemical as well as with classical molecular dynamics methods. The structural zinc site is composed of four closely spaced cysteine ligands (Cys97, Cys100, Cys103 and Cys111 in the amino acid sequence) positioned in an almost symmetric tetrahedron around the Zn ion. A recent study showed that the interaction between zinc and cysteine is governed by primarily an electrostatic contribution with an additional covalent contribution to the binding.[11]



In humans, ADH exists in multiple forms as a dimer and is encoded by at least seven different genes. There are five classes (I-V) of alcohol dehydrogenase, but the hepatic form that is primarily used in humans is class 1. Class 1 consists of A,B, and C subunits that are encoded by the genes ADH1A, ADH1B, and ADH1C.[12] The enzyme is contained in the lining of the stomach and in the liver. It catalyzes the oxidation of ethanol to acetaldehyde:


This allows the consumption of alcoholic beverages, but its evolutionary purpose is probably the breakdown of alcohols naturally contained in foods or produced by bacteria in the digestive tract. Others believe that its evolutionary purpose is involved in vitamin A metabolism, as alcohols are relatively 'empty' calories, providing little net nutritional benefit.[citation needed]

alcohol dehydrogenase 1A,
α polypeptide
Symbol ADH1A
Alt. symbols ADH1
Entrez 124
HUGO 249
OMIM 103700
RefSeq NM_000667
UniProt P07327
Other data
EC number
Locus Chr. 4 q23
alcohol dehydrogenase 1B,
β polypeptide
Symbol ADH1B
Alt. symbols ADH2
Entrez 125
HUGO 250
OMIM 103720
RefSeq NM_000668
UniProt P00325
Other data
EC number
Locus Chr. 4 q23
alcohol dehydrogenase 1C,
γ polypeptide
Symbol ADH1C
Alt. symbols ADH3
Entrez 126
HUGO 251
OMIM 103730
RefSeq NM_000669
UniProt P00326
Other data
EC number
Locus Chr. 4 q23

Alcohol dehydrogenase is also involved in the toxicity of other types of alcohol: for instance, it oxidizes methanol to produce formaldehyde and ethylene glycol to ultimately yield glycolic and oxalic acids. Humans have at least six slightly different alcohol dehydrogenases. Each is a dimer (i.e., consists of two polypeptides), with each dimer containing two zinc ions Zn2+. One of those ions is crucial for the operation of the enzyme: it is located at the catalytic site and holds the hydroxyl group of the alcohol in place.

Alcohol dehydrogenase activity varies between men and women, between young and old, and among populations from different areas of the world. For example, young women are unable to process alcohol at the same rate as young men because they do not express the alcohol dehydrogenase as highly. Though, the inverse is true amongst the middle-aged.[13] The level of activity may not only be dependent on level of expression but due to allelic diversity among the population. These allelic differences have been linked to region of origin. For example, populations from Europe have been found to express an allele for the alcohol dehydrogenase gene that makes it much more active than those found in populations from Asia or the Americas.[citation needed]

This may be a correlating evolution with the rise of aldehyde dehydrogenase, which has been suggested as one of the more recognizable recent evolutionary changes in humans (along with lactose tolerance) - in order to make water safe in cities too dense to use springs, Europeans fermented alcoholic (and hence antiseptic) beverages, while East Asians typically boiled their water (creating, among other things, tea). The Europeans' relatively greater alcohol consumption increased selection for those who didn't suffer from violent alcohol flush response in European populations.[citation needed]

The human genes that encode class II, III, IV, and V alcohol dehydrogenases are ADH4, ADH5, ADH7, and ADH6, respectively.

alcohol dehydrogenase 4
(class II), π polypeptide
Symbol ADH4
Entrez 127
HUGO 252
OMIM 103740
RefSeq NM_000670
UniProt P08319
Other data
EC number
Locus Chr. 4 q22
alcohol dehydrogenase 5
(class III), χ polypeptide
Symbol ADH5
Entrez 128
HUGO 253
OMIM 103710
RefSeq NM_000671
UniProt P11766
Other data
EC number
Locus Chr. 4 q23
alcohol dehydrogenase 6
(class V)
Symbol ADH6
Entrez 130
HUGO 255
OMIM 103735
RefSeq NM_000672
UniProt P28332
Other data
EC number
Locus Chr. 4 q23
alcohol dehydrogenase 7
(class IV), μ or σ polypeptide
Symbol ADH7
Entrez 131
HUGO 256
OMIM 600086
RefSeq NM_000673
UniProt P40394
Other data
EC number
Locus Chr. 4 q23-q24

Yeast and bacteria

Unlike humans, yeast and bacteria do not ferment glucose to lactate. Instead, they ferment it to ethanol and CO2. The overall reaction can be seen below.

Glucose + 2 ADP + 2 Pi → 2 ethanol + 2 CO2 + 2 ATP + 2 H2O[14]

In yeast and many bacteria, alcohol dehydrogenase plays an important part in fermentation: pyruvate resulting from glycolysis is converted to acetaldehyde and carbon dioxide, and the acetaldehyde is then reduced to ethanol by an alcohol dehydrogenase called ADH1. The purpose of this latter step is the regeneration of NAD+, so that the energy-generating glycolysis can continue. Humans exploit this process to produce alcoholic beverages, by letting yeast ferment various fruits or grains. It is interesting to note that yeast can produce and consume their own alcohol.

The main alcohol dehydrogenase in yeast is larger than the human one, consisting of four rather than just two subunits. It also contains zinc at its catalytic site. Together with the zinc-containing alcohol dehydrogenases of animals and humans, these enzymes from yeasts and many bacteria form the family of "long-chain"-alcohol dehydrogenases.

Brewer's yeast also has another alcohol dehydrogenase, ADH2, which evolved out of a duplicate version of the chromosome containing the ADH1 gene. ADH2 is used by the yeast to convert ethanol back into acetaldehyde, and it is only expressed when sugar concentration is low. Having these two enzymes allows yeast to produce alcohol when sugar is plentiful (and this alcohol then kills off competing microbes), and then continue with the oxidation of the alcohol once the sugar, and competition, is gone.[15]

Alcohol Dehydrogenase from Brewer's Yeast can be easily purified as follows: Using a 25 mM Pyrophosphate Buffer containing 5 mM Zinc Chloride, 5 mM EDTA, and 0.5 mg/ml BSA, disrupt yeast cells with a bead mill. Perform an ammonium sulfate precipitation at 40%, and discard the pellet. Perform a second ammonium sulfate precipitation at 70%, resuspend the resulting pellet in 1 ml of buffer. Run on a Sephadex G-100 column; ADH will elute in first few fractions. The sample can then be concentrated with a high concentration of ammonium sulfate as there should be few other proteins present.


A third family of alcohol dehydrogenases, unrelated to the above two, are iron-containing ones. They occur in bacteria, and an apparently inactive form has also been found in yeast[citation needed]. In comparison to enzymes the above families, these enzymes are oxygen-sensitive.

Other types

A further class of alcohol dehydrogenases belongs to quinoenzymes and requires quinoid cofactors (e.g., pyrroloquinoline quinone, PQQ) as enzyme-bound electron acceptors. A typical example for this type of enzyme is methanol dehydrogenase of methylotrophic bacteria.


In fuel cells, alcohol dehydrogenases can be used to catalyze the breakdown of fuel for an ethanol fuel cell. Scientists at Saint Louis University have used carbon-supported alcohol dehydrogenase with poly(methylene green) as an anode, with a nafion membrane, to achieve about 50 μA/cm2.[16]

In biotransformation, alcohol dehydrogenases are often used for the synthesis of enantiomerically pure stereoisomers of chiral alcohols. In contrast to the chemical process, the enzymes yield directly the desired enatiomer of the alcohol by reduction of the corresponding ketone.

Clinical significance


There have been studies showing that ADH may have an influence on the dependence on ethanol metabolism in alcoholics. Researchers have tentatively detected a few genes to be associated with alcoholism. If the variants of these genes encode slower metabolizing forms of ADH2 and ADH3, there is increased risk of alcoholism. The studies have found that mutations of ADH2 and ADH3 are related to alcoholism in Asian populations. However, research continues in order to identify the genes and their influence on alcoholism.[17]

Drug dependence

Drug dependence is another problem associated with ADH, which researchers think might be linked to alcoholism. One particular study suggests that drug dependence has seven ADH genes associated with it. These results may lead to treatments that target these specific genes. However, more research is necessary.[18]

Picture gallery

Alcohol Dehydrogenase.  
Horse LADH (Liver Alcohol Dehydrogenase)  
Structural zinc site motif in alcohol dehydrogenase  

See also


  1. ^ PDB 1m6h; Sanghani PC, Robinson H, Bosron WF, Hurley TD (September 2002). "Human glutathione-dependent formaldehyde dehydrogenase. Structures of apo, binary, and inhibitory ternary complexes". Biochemistry 41 (35): 10778–86. PMID 12196016. 
  2. ^ a b Danielsson O, Jörnvall H (October 1992). ""Enzymogenesis": classical liver alcohol dehydrogenase origin from the glutathione-dependent formaldehyde dehydrogenase line". Proc. Natl. Acad. Sci. U.S.A. 89 (19): 9247–51. PMID 1409630. 
  3. ^ a b Persson B, Hedlund J, Jörnvall H (December 2008). "Medium- and short-chain dehydrogenase/reductase gene and protein families : the MDR superfamily". Cell. Mol. Life Sci. 65 (24): 3879–94. doi:10.1007/s00018-008-8587-z. PMID 19011751. 
  4. ^ Negelein E, Wulff HJ (1937)). Biochem. Z. 293: 351. 
  5. ^ Theorell H, McKee JS (October 1961). "Mechanism of action of liver alcohol dehydrogenase". Nature 192: 47–50. PMID 13920552. 
  6. ^ Jörnvall H, Harris JI (April 1970). "Horse liver alcohol dehydrogenase. On the primary structure of the ethanol-active isoenzyme". Eur. J. Biochem. 13 (3): 565–76. PMID 5462776. 
  7. ^ Brändén CI, Eklund H, Nordström B, Boiwe T, Söderlund G, Zeppezauer E, Ohlsson I, Akeson A (August 1973). "Structure of liver alcohol dehydrogenase at 2.9-angstrom resolution". Proc. Natl. Acad. Sci. U.S.A. 70 (8): 2439–42. PMID 4365379. 
  8. ^ Hellgren M (2009). Enzymatic studies of alcohol dehydrogenase by a combination of in vitro and in silico methods, Ph.D. thesis. Stockholm, Sweden: Karolinska Institutet. pp. 70. ISBN 978-91-7409-567-8. 
  9. ^ a b Sofer W, Martin PF (1987). "Analysis of alcohol dehydrogenase gene expression in Drosophila". Annual Review of Genetics 21: 203–25. doi:10.1146/ PMID 3327463. 
  10. ^ a b c d Hammes-Schiffer S, Benkovic SJ (2006). "Relating protein motion to catalysis". Annual Review of Biochemistry 75: 519–41. doi:10.1146/annurev.biochem.75.103004.142800. PMID 16756501. 
  11. ^ Erik G. Brandt, Mikko Hellgren, Tore Brinck, Tomas Bergman and Olle Edholm (2009). "Molecular dynamics study of zinc binding to cysteines in a peptide mimic of the alcohol dehydrogenase structural zinc site". Phys. Chem. Chem. Phys. (PCCP) 11 (6): 975–83. PMID 19177216. 
  12. ^ Sultatos LG, Pastino GM, Rosenfeld CA, Flynn EJ (March 2004). "Incorporation of the genetic control of alcohol dehydrogenase into a physiologically based pharmacokinetic model for ethanol in humans". Toxicological Sciences : an Official Journal of the Society of Toxicology 78 (1): 20–31. doi:10.1093/toxsci/kfh057. PMID 14718645. 
  13. ^ Parlesak A, Billinger MH, Bode C, Bode JC (2002). "Gastric alcohol dehydrogenase activity in man: influence of gender, age, alcohol consumption and smoking in a caucasian population". Alcohol and Alcoholism (Oxford, Oxfordshire) 37 (4): 388–93. PMID 12107043. 
  14. ^ Cox, Michael; Nelson, David R.; Lehninger, Albert L (2005). Lehninger principles of biochemistry. San Francisco: W.H. Freeman. pp. 180. ISBN 0-7167-4339-6. 
  15. ^ Coghlan A (2006-12-23). "Festive special: The brewer's tale - life". New Scientist. Retrieved 2009-04-27. 
  16. ^ Moore CM, Minteer SD, Martin RS (February 2005). "Microchip-based ethanol/oxygen biofuel cell". Lab on a Chip 5 (2): 218–25. doi:10.1039/b412719f. PMID 15672138. 
  17. ^ Sher KJ, Grekin ER, Williams NA (2005). "The development of alcohol use disorders". Annual Review of Clinical Psychology 1: 493–523. doi:10.1146/annurev.clinpsy.1.102803.144107. PMID 17716097. 
  18. ^ Luo X, Kranzler HR, Zuo L, Wang S, Schork NJ, Gelernter J (February 2007). "Multiple ADH genes modulate risk for drug dependence in both African- and European-Americans". Human Molecular Genetics 16 (4): 380–90. doi:10.1093/hmg/ddl460. PMID 17185388. 

External links

  • PDBsum has links to three-dimensional structures of various alcohol dehydrogenases contained in the Protein Data Bank
  • ExPASy contains links to the alcohol dehydrogenase sequences in Swiss-Prot, to a Medline literature search about the enzyme, and to entries in other databases.
  • Radio Free Genome created a musical score from a sequence of alcohol dehydrogenase. MP3 audio version and an open source version of the software used to create it is available.

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