Carbohydrate Fermentation

Department of microbiology
Presented By
Javairia
Sofia
Table of content

– Introduction
– Bio-oxidative pathways
– Fermentation
– Carbohydrate fermentation
– The Embden-Meyerhof pathway
– Carbohydrate non-fermenters
– Conclusion
Introduction
– A carbohydrate is a biomolecule consisting of carbon, hydrogen and oxygen
atoms
– Usually with a hydrogen–oxygen atom ratio of 2:1
– Carbohydrates are the sugars, starches and fibers found in fruits, grains,
vegetables and milk products
– carbohydrates — one of the basic food groups — are important to a healthy
diet
 Cont…
– Carbohydrate chains come in different lengths
– Biologically important carbohydrates belong to three categories: monosaccharides, disaccharides, and
polysaccharides
– Monosaccharides (mono- = “one”; sacchar- = “sugar”) are simple sugars, the most common of which is glucose.
Monosaccharides have a formula of (CH2​O)n​and they typically contain three to seven carbon atoms.
 Cont…
– Disaccharides (di- = “two”) form when two monosaccharides join together via a dehydration reaction, also
known as a condensation reaction. Releasing a molecule of water and forming a covalent bond known as a
glycosidic linkage.
– A long chain of monosaccharides linked by glycosidic bonds is known as a polysaccharide (poly- = “many”).
The chain may be branched or unbranched and may contain different types of monosaccharides from
hundreds to thousands. Starch, glycogen, cellulose, and chitin are some major examples of polysaccharides
important in living organisms.
 Bio-oxidative pathways
– Most microorganisms obtain their energy through a series of orderly and integrated enzymatic reactions leading
to the biooxidation of a substrate, frequently a carbohydrate.
– The major pathways by which this is accomplished are shown in
– Aerobic: Bio-oxidations in which molecular oxygen can serve as the final electron acceptor
– Anaerobic: Bio-oxidations in which inorganic ions other than oxygen, such as NO3 – or SO4 2–, can serve as the
final electron acceptors
– Fermentation: A bio-oxidative process not requiring oxygen in which an organic substrate serves as the final
electron acceptors
 Fermentation
– Fermentation is a metabolic process that produces chemical changes in organic substrates through the
action of enzymes
– Fermentation involves a complex system of reactions brought about by microorganism that may be present
simultaneously.
– There are different types of fermentation resulting from the action of yeast and other microorganisms such
as lactic acid bacteria
– Fermentation is a process used to produce wine, beer, yogurt and other products.
Cont…
 Carbohydrate fermentation
– Organisms use carbohydrates differently depending on their enzyme complement.
– Some organisms are capable of fermenting sugars such as glucose anaerobically, while others use the
aerobic pathway.
– Others, facultative anaerobes, are enzymatically competent to use both aerobic and anaerobic pathways
– Organisms lack the ability to oxidize glucose by either. In this exercise the fermentative pathways are of
prime concern
 Cont…
– In fermentation, substrates such as carbohydrates and alcohols undergo anaerobic breakdown and produce
an organic acid (for example, lactic, formic, or acetic acid) along with gases such as hydrogen or carbon
dioxide.
– Fermentation is best described by considering the degradation of glucose by way of the Embden-Meyerhof
pathway, also known as the glycolytic pathway
– Facultative anaerobes are usually the socalled fermenters of carbohydrates.
 The Embden-Meyerhof pathway
– One mole of glucose is converted into two moles of pyruvic acid, which is the major intermediate compound
produced by glucose degradation.
– Metabolism of pyruvate is not the same for all organisms, and a variety of end products result that define
their different fermentative capabilities.
 Procedure
– Fermentative degradation under anaerobic conditions is carried out in a fermentation broth tube containing
a Durham tube, an inverted inner vial for the detection of gas production.
 Cont…
– A typical carbohydrate fermentation medium contains:
– 1. Nutrient broth ingredients for the support of the growth of all organisms.
– 2. A specific carbohydrate that serves as the substrate for determining the organism’s fermentative
capabilities.
– 3. The pH indicator phenol red, which is red at a neutral pH (7) and changes to yellow at a slightly acidic pH of
6.8, indicating that slight amounts of acid will cause a color change.
– All cultures should be observed within 48 hours.
 Identification
– Carbohydrates that have been fermented with the production of acidic wastes will cause the phenol red to
turn yellow, thereby indicating a positive reaction
 Cont…
– In some cases, acid production is accompanied by the evolution of a gas (CO2) that will be visible as a
bubble in the inverted tube.
– Cultures that are not capable of fermenting a carbohydrate substrate will not change the indicator, and the
tubes will appear red; there will not be a concomitant evolution of gas. This is a negative reaction.
 Carbohydrate non-fermenters

– The lack of carbohydrate fermentation by some organisms should not be considered as absence of growth.
– The organisms use other nutrients in the medium as energy sources. Such as peptones present in nutrient
broth.
– Peptones can be degraded by microbial enzymes  amino acids  enzymatically converted by oxidative
deamination to ketoamino acids  metabolized through the Krebs cycle for energy production.
– These reactions liberate ammonia, which accumulates in the medium, forming ammonium hydroxide (NH4OH)
and producing an alkaline environment. When this occurs, the phenol red turns to a deep red in the basic
medium.
 Conclusion
– Carbohydrates are the main source of energy for living organisms
– Fermentation is a metabolic bio-oxidative chemical and anaerobic process used for the breakdown of
carbohydrates
– Carbohydrate fermentation is carried mainly by
“The Embden-Meyerhof pathway”
– The identification is carried by using indicator in the medium