FERMENTATION

• Classical Biotechnology
• Humans have been using this technology
for centuries
• Involves harnessing the wastes of bacteria
and/or yeast for products that humans
consume
• Big business
 Cellular Respiration: the process of using
glucose to make energy (ATP) for the cell.

Aerobic Cellular Respiration (requires oxygen)

enzymes
6O2 + C6H12O6 6CO2 + 6H2O
oxygen glucose carbon dioxide water
ENERGY transfer
between enzymes,
other molecules
ADP + Pi ATP
•Aerobic Cellular Respiration takes place
in the mitochondria of cells.

•It can provide up to 38 molecules of ATP

per molecule of glucose.
Anaerobic Cellular Respiration (without oxygen)

also known as Fermentation

Alcoholic Fermentation:

C6H12O6 2CO2 + 2C2H5OH

glucose carbon dioxide ethanol

•provides 2 molecules ATP per glucose

•done by yeast
 Products of Alcoholic Fermentation

don’t drink alcohol

Making Rootbeer: 6 simple steps
• Heat Water to ~40 degrees C (yeast like it)
• Add sugar and dissolve
• Add root beer extract
• Add yeast
• Ferment @ ~27 degrees C
• Chill and enjoy!
 Lactic Acid Fermentation:

Glucose carbon dioxide + lactic acid

•provides 2 molecules ATP per glucose

•done by muscle cells
•done by bacteria cells
Products of Lactic Acid Fermentation
Other products of fermentation - some are
fermented by both yeast and bacteria

Idli, Dosas, Kimchee, Sausage, Kefir,

sauerkraut, miso, tempeh, tamari,
chutney
More products of fermentation
 Yogurt
Yogurt-like products have been made for millenia across Eastern Europe,
North Africa, Central Asia and India.

Contains bacteria that are “thermophilic” = heat loving

 Two main types of Lactic Acid Bacteria
(Identified around the year 1900):
Lactobacillus
• meaning “milk” and “rod”
• over 50 different species
• found on plants and in the digestive system of animals such as cows
and humans.
Lactococcus
• meaning “milk” and “sphere” because of its shape
• found primarily on plants
• less common than lactobacillus
Traditional spontaneously fermented milks contain species
that can reside in the human digestive tract:
• Lactobacillus fermentum,
• L. casei
• L brevis
• L plantarum (from picked vegetables)
• L acidophilus
 Commercial Yogurt
Contains 2 species of bacteria specialized to grow well in milk (but can’t
survive inside the human body):

First, Next,
Streptococcus Lactobacillus
thermophilus bulgaricus is
is more active, more acid
then slows tolerant and
down when takes over until
acidity acidity >1%
reaches 0.5%

These bacteria work in symbiosis. Each bacterium stimulates the growth

of the other => acidifies the milk more rapidly than either partner on its
own.
 Milk is

•Water
•Protein (casein and whey)

•Fat

•Sugar (lactose)
•Vitamins
• Minerals
 How Does Milk Turn Into Yogurt?

Lactic acid Bacteria

Lactose -------------------------> Lactic Acid
(Milk sugar)

Acid causes casein (milk

protein) to denature and
hold water into a
semi-solid gel = yogurt
 Milk Yogurt
Casein protein micelles
(bundles)
Bacteria produce acid
10-7 meters in diameter

Acid causes
Casein bundles to
fall apart into
separate casein
molecules.
These rebind to
each other in a
Fat network that traps
globule water.
=> makes a gel
 Making Yogurt in 4 Simple Steps
1. Start with Cow, Sheep, or Goat milk.
2. Heat milk to 80 °C. Two purposes:
• destroy existing bacteria
• “condition” the proteins = begins the denaturing process
(a whey protein molecule binds to a casein molecule which disrupts
the casein bundles allowing them to make short branched micelle
chains)
3. Cool milk to 40 °C and innoculate with bacteria
4. Incubate at 30 °C to 45 °C
Casein before heat Casein after heat
pre-treatment: pre-treatment: Casein after acid:
 Incubation Temperature
• 40-45 °C takes 2-3 hours

Produces a coarse protein network with thick strands give

firmness but easily leak whey (a process called syneresis - the
separation of liquid from the gel)

• 30 °C takes 18 hours

Produces a finer more branched delicate network that holds the

liquid whey