MICROBIAL METABOLISM

1. CARBOHYDRATE CATABOLISM
2. FERMENTATION OF CARBOHYDRATES
3. PROTEIN CATABOLISM
4. RESPIRATION
5. RAPID IDENTIFICATION METHODS

MICROBIAL METABOLISM

Metabolism is the sum of the chemical reactions in

an organism.
Catabolism is the energy-releasing processes.
Anabolism is the energy-using processes.

MICROBIAL METABOLISM

A metabolic pathway is a sequence of enzymatically

catalyzed chemical reactions in a cell.

Metabolic pathways are determined by enzymes.
Enzymes are encoded by genes.

MICROBIAL METABOLISM

The majority of enzymes function inside a cell-that

is, they are endoenzymes.

Many

bacteria make some enzymes, called

exzoenzymes, that are released from the cell to
catalyze reactions outside of the cell.

1. CARBOHYDRATE CATABOLISM
Most bacteria catabolize carbohydrates for carbon

and energy.
Carbohydrates can be classified based on size:
monosaccharides, oligosaccharides, polysaccharides.
Exzoezymes are mainly hdyrolytic enzymes that

leave the cell and break down, by the addition of

water, large subrates into smaller components,
which can be transported into the cell.

1. CARBOHYDRATE CATABOLISM
Amylase hydrolyzes the polysaccharide starch into

smaller carbohydrates.
In the laboratory, the presence of an exzoenzyme is

determined by looking for a change in the substrate

outside of a bacterial colony.
starch

Amylase
H2O

glucose+glucose+

1. CARBOHYDRATE CATABOLISM
Procedure of Starch Hydrolysis Test

With a marker, divide the petri into three sector.

2. Streak the bacteria (Bacillus, E.coli and
Pseudomonas)
3. Incubate the plate for 24h.
4. After the incubation, flood the plate with Grams
iodine. Areas of starch hydrolysis will appear clear ,
while unchanged starch will stain dark blue.
1.

2. FERMANTATION OF CARBOHYDRATES
Releases energy from fermentation of organic molecules

Does not require oxygen

Does not use the Krebs cycle or ETC
Most bacteria can ferment the glucose without using

oxygen.
The metabolic end-products of fermantation are small
organic molecules, usually organic acids. Some bacteria
produce gases from the fermantation of carbohydrates.

2. FERMANTATION OF CARBOHYDRATES

2. FERMANTATION OF CARBOHYDRATES

A fermantation tubes (duraham tube) is used to detect

acid and gas production from carbohydrates.
Acid-base indicator phenol red
inverted tube trap gas

3. PROTEIN CATABOLISM (urease)

3. PROTEIN CATABOLISM (urease)

Urea

urease
H2O

Ammonia (NH3) + CO2

Bacteria can hydrolize the peptides or polypeptides to release

amino acids.
They use the amino acid as carbon and energy sources.
Urea is waste product of protein digestion

3. PROTEIN CATABOLISM (indole)

Tryptophan

hydrolysis-Some
bacteria
split
tryptophan into indole and pyruvic acid using the
hydrolase called tryptophanase.
Indole can be detected with Kovac's reagent (Indole
reagent).

4. Respiration
Catalase Test
Microorganisms able to live in oxygenated environments

produce enzymes which neutralize toxic forms of oxygen.

One such enzyme is catalase, which breaks hydrogen
peroxide into water and molecular oxygen. Organisms
which produce catalase will bubble when placed into
hydrogen peroxide.

4. Respiration
Oxidase Test
The oxidase test is a test
used in microbiology to
determine if a
bacterium produces certain
cytochrome c oxidases. It
uses disks impregnated with
a reagent.

Gram Test (3% KOH)

The KOH test is based on differences in the

chemistry of the bacterial cell wall. The cell wall of

gram-negative bacteria is easily disrupted when
exposed to dilute alkali solutions.
When the cell walls are disrupted, the suspension
in KOH becomes viscous due to the release of
relatively unfragmented threads of deoxyribonucleic
acid.

5. Rapid Identification Methods

Identify the bacteria quickly and accurately.
Provide large number of results from one

inoculation.

Api 20E strip

Biolog

Vitek 2 Compact

Experiments in this lab

Exp 4: Respiration
Oxidase (E.coli, Pseudomonas)
Catabolism
Starch agar (E.coli, Bacillus, Catalase (Bacillus,
Staphylococcus)
Pseudomonas)
Exp 5: Rapid Identification
Exp 2: Protein Catabolism 1
Methods
Urea agar (Pseudomonas
Citrate (E.coli and
and Proteus vulgaris)
Enterobacter)
Exp 3: Protein Catabolism 2 Exp 6: Gram test by KOH
Indole Test (E.coli, Proteus 3% KOH (E.coli and Bacillus)
vulgaris)
Exp 1: Carbohydrate