Eduard Buchner

(1860-1917) 1897 found fermentation in broken yeast cells 1907 Nobel Prize in Chemistry

The whole pathway in yeast and muscle cell were elucidated by

Arthur Harden 1865-1940

Glycolysis
Glycolysis is an almost universal central pathway of glucose catabolism, the pathway with the largest flux of carbon in most cells. In some mammalian tissues (erythrocytes, renal medulla, brain, sperm), the glycolytic breakdown of glucose is the sole source of metabolic energy.

Glycolysis
Some of the starch-storing tissues, like potato tubers, and some aquatic plants derive most of their energy from glycolysis. Many anaerobic microorganisms are entirely dependent on glycolysis.

1. phosphorylation of glucose

2. Isomerization of glucose 6phosphate

Phosphohexose isomerase reaction

by an active-site His residue

Glu

3. Phosphorylation of fructose 6phosphate: the first committed step in glycolysis

PFK-1 is named so because there is another enzyme catalyzes a similar reaction

In some bacteria, protists and (all) plants, a pyrophosphate-dependent phosphofructokinase (PFP) also catalyzes this reaction in a reversible way

4. Cleavage of fructose 1,6bisphosphate

Class I aldolases form Schiff base intermediate during sugar cleavage reaction
Class I aldolases were found in animals and plants. Class II aldolases (fungi and bacteria) do not form the Schiff base and require a zinc ion to catalyze reaction.

5. Interconversion of the triose phosphate

Dihydroxyacetone phosphate and glyceraldehyde 3-phosphate become indistinguishable after triose phosphate isomerase reaction

6. Oxidation of glyceraldehyde 3phosphate to 1,3-bisphosphoglycerate

The glyceraldehyde 3-phosphate dehydrogenase reaction

Heavy metal ion such as Hg2+ will react with Cys residue, hence irreversibly inhibits the enzyme.

hemiacetal

7. Phosphoryl transfer from 1,3bisphosphoglycerate to ADP

Glyceraldehyde 3-phosphate dehydrogenase and Phosphoglycerate kinase are coupled in vivo

Glyceraldehyde 3-phosphate dehydrogenase catalyzes an endergonic reaction while phosphoglycerate kinase catalyzes an exergonic reaction. When these two reactions are coupled (which happens in vivo), the overall reaction is exergonic.

The formation of ATP by phosphoryl group transfer from a substrate is referred to as a substrate-level phosphorylation

Substrate-level phosphorylation
soluble enzymes chemical intermediates

Respiration-linked phosphorylation Photophosphorylation

membrane-bound enzymes transmembrane gradients of protons

8. Conversion of 3phosphoglycerate to 2phosphoglycerate

The phosphoglycerate mutase reaction

2,3-Bisphosphoglycerate (BPG)
The concentration of BPG is usually low in most of the tissues except erythrocytes (up to 5 mM). Function of BPG in erythrocytes is to regulate the affinity of hemoglobulin to O2.

9. Dehydration of 2phosphoglycerate to phosphoenolpyruvate

10. Transfer of the phosphoryl group from phosphoenolpyruvate to ADP

Glucose + 2ATP + 2NAD+ + 4ADP + 2Pi 2 pyruvate + 2ADP + 2NADH + 2H+ + 4ATP + 2H2O
Glucose + 2ADP + 2NAD+ + 2Pi 2 pyruvate + 2ATP + 2NADH + 2H+ NADH mitochondriaATP

NAD+ (nicotinamide adenine dinucleotide) is the active form of niacin

Niacin
Niacin is the common name for nicotinamide and nicotinic acid. Nicotinic acid is the common precursor for NAD+ and NADP+ biosynthesis in cytosol.

Functions of

+ NAD

and

+ NADP

Both NAD+ and NADP+ are coenzymes for many dehydrogenases in cytosol and mitochondria NAD+ is involved in oxidoreduction reactions in oxidative pathways. NADP+ is involved mostly in reductive biosynthesis.

Niacin deficiency: pellagra

Weight loss, digestive disorders, dermatitis, dementia

Niacin deficiency
Because niacin is present in most of the food and NAD+ can also be produced from tryptophan (60 grams of trptophan 1 gram of NAD+), so it is not often to observe niacin deficiency. However, niacin deficiency can still be observed in areas where maize is the main carbohydrate source because maize only contain niacytin, a bound unavailable form of niacin. Pre-treated maize with base will release the niacin from niacytin.

Niacin deficiency
Areas where sorghum is the main carbohydrate source will also observe niacin deficiency if niacin uptake is not being watched carefully. Sorghum contains large amount of leucine, which will inhibit quinolinate phosphoribosyl transferase (QPRT), an enzyme involved in NAD+ biosynthesis from tryptophan. Vitamin B6 deficiency can also lead to niacin deficiency because pyridoxal phosphate is a coenzyme in NAD+ biosynthesis from tryptophan.

ISONIAZID A Commonly Used Medication for HIV & AIDS Patients

Drug:
Classification: Indication:

ISONIAZID
Antimycobacterial Infection with, or disease from, mycobacterium tuberculosis

Feeder pathways for glycolysis

Glycogen and starch are degraded by phosphorolysis

Glycogen and starch can be mobilized for use by a phosphorolytic reaction catalyzed by glycogen/starch phosphorylase. This enzyme catalyze an attack by Pi on the (a14) glycosidic linkage from the nonreducing end, generating glucose 1phosphate and a polymer one glucose unit shorter.

Branch point (a16) is removed by debranching enzyme

Glucose 1-phosphate is converted to G-6-P by phosphoglucomutase by the same mechanism observed in phosphoglycerate mutase reaction

Digestion of dietary polysaccharides

Digestion begins in the mouth with salivary aamylase hydrolyze (attacking by water) the internal glycosidic linkages. Salivary a-amylase is then inactivated by gastric juice; however pancreatic a-amylase will take its place at small intestine. The products are maltose, maltotriose, and limit dextrins (fragments of amylopectin containing a16 branch points.

Digestion of dietary disaccharides

Disaccharides must be hydrolyzed to monosaccharides before entering cells. Dextrin + nH2O n D-glucose dextrinase Maltose + H2O 2 D-glucose maltase Lactose + H2Olactase D-galactose + D-glucose Sucrose + H2O D-fructose + D-glucose sucrase Trehalose + H2O 2 D-glucose trehalase

Lactose intolerance

Lactose intolerance is due to the disappearance after childhood of most or all of the lactase activity of the intestinal cells.

Lactose intolerance
Undigested lactose will be converted to toxic products by bacteria in large intestine, causing abdominal cramps and diarrhea.

Fructose metabolism in muscle and kidney

Fructose metabolism in liver

Triose phosphate isomerase

In liver, the enzyme fructokinase catalyze the phosphorylation of fructose to form fructose 1-phosphate.

Galactose metabolism

Galactose is phosphorylated by galactokinase in the liver. Then galactose 1-phosphate is converted to glucose 1-phosphate by a series of reactions.

Galactose metabolism
The conversion of galactose 1-P to glucose 1-P (epimerization) requires uridine diphosphate (UDP) as a coenzyme-like carrier of hexose groups.

Galactosemia
inability to metabolize galactose due to lack of 1. UDP-glucose galactose 1-phosphate uridylyltransferase (classical galactosemia) 2. UDP-glucose 4-epimerase 3. Galactokinase Among these, deficiency of either 1 or 2 is more severe (1 is the most severe).

Galactosemia
Deficiency of transferase (or epimerase) will result in poor growth, speech abnormality, mental deficiency, and (fatal) liver damage even when galactose is withheld from the diet.

Galactosemia patients develop cataracts by deposition of galactitol in the lens

Mannose metabolism
Mannose + ATP mannose 6-phosphate hexokinase +ADP mannose 6-phosphate fructose 6-phosphate
phosphomannose isomerase

Fermentation
Fermentation is referring to the process when no oxygen is consumed or no change in the concentration of NAD+ or NADH during energy extraction.

Fermentation

Under hypoxic conditions, oxidative phosphorylation will be the first to stop. Then citric acid cycle will come to a halt due to inhibition effect from NADH. As a result, glycolysis will be the only metabolic pathway that is available to energy production during hypoxia.

Fermenation
However, the oxidation of glyceraldehyde 3phosphate consumes NAD+ that will not be regenerated under hypoxic condition because oxidative phosphorylation is not available.

The purpose of fermentation is to regenerate NAD+

In order to continue regenerating NAD+, cells come up a strategy. During fermentation, NAD+ is regenerated during the reduction of pyruvate, the product of glycolysis.

Lactate fermentation
glycolysis

Lactate is recycled in the liver (Cori cycle)

Carl and Gerty Cori, 1947 Nobel Prize in Physiology and Medicine

Lactate fermentation only happened in larger animals

Most small vertebrates and moderate size running animals have circulatory systems that can carry oxygen to their muscles fast enough to avoid having to use muscle glycogen anaerobically.

http://www.mountain-research.org/CV/coelacanth.jpg

http://www.anac.8m.net/Images/coelacanth.jpg

Deep sea fish (below 4,000 m or more) coelacanth uses anaerobic metabolism exclusively. The lactate produced is excreted directly. Some marine vertebrates can do ethanol fermentation.

Ethanol fermentation

Yeast and other microorganisms ferment glucose to ethanol and CO2. Pyruvate is first decarboxylated by pyruvate decarboxylase, which is absent in vertebrate tissues and in other organisms that carry out lactic acid fermentation. Acetaldehyde is the product of this reaction.

Pyruvate decarboxylase
The decarboxylation of pyruvate by pyruvate decarboxylase produces CO2, which is the reason why champagne is bubbling.

Thiamine pyrophosphate (TPP) is the coenzyme of pyruvate decarboxylase

Thiamine pyrophosphate is derived from vitamin B1 (thiamine). Lack of vitamine B1 will lead to beriberi (edema, pain, paralysis, death).

TPP plays an important role in the cleavage of bonds adjacent to a carbonyl group.
The thiazolium ring of TPP acts as an electron sink to facilitates decarboxylation reaction.

Alcohol dehydrogenase catalyze the second step of ethanol fermentation

Alcohol dehydrogeanse reduces acetaldehyde, producing NAD+ and ethanol. This enzyme is present in many organisms that metabolize ethanol, including human.

Fermentation has commercial values

Bacteria like Lactobacillus bulgaricus (yogurt) and Propionibacterium freudenreichii (swiss cheese) ferments milk to produce lactic acid or propionic acid and CO2.

Dr. Chaim Weizmann 1874-1952 First President of Israel Found butanol and acetone fermentation in Clostridium acetobutyricum

Industrial fermentation is done in huge close vats

Fermentors are huge closed vats in which temperature and access to air are adjusted to favor the multiplication of the desired microorganism. Some even immobilize the cells in an inert support so no effort is required to separate microorganisms from products after fermentation is completed.