Extremophiles

Life on the edge

Life at High Temperatures, Thomas M. Brock

Extremophiles

Images from NASA, http://pds.jpl.nasa.gov/planets/

Goals

Overview of Extremophiles
Review some biology
Give some applications
Motivate you to study Microbiology!!

IntroductiontoExtremophiles

What are they?

Microbes living where nothing else can

How do they survive?

Extremozymes

Why are they are interesting?

Extremes fascinate us

Life on other planets

Practical applications are interesting

Interdisciplinary lessons
Genetic Prospecting

Extremo phile

Definition - Lover of extremes

History

Suspected about 30 years ago

Known and studied for about 20 years

Temperature extremes

boiling or freezing, 1000C to -10C (212F to

30F)

Chemical extremes

vinegar or ammonia (<5 pH or >9 pH)

highly salty, up to ten times sea water

How we sterilize & preserve foods today

Extreme Temperatures

Thermophiles - High temperature

Thermal vents and hot springs
May go hand in hand with chemical extremes

Psychrophiles - Low temperature

Arctic and Antarctic
1/2 of Earths surface is oceans between 10C & 40C
Deep sea 10C to 40C
Most rely on photosynthesis

Thermophiles

Obsidian Pool,
Yellowstone National
Park
Hydrothermal Vents

Psychrophiles

Chemical Extremes

Acidophiles - Acidic
Again thermal vents and some hot springs

Alkaliphiles - Alkaline
Soda lakes in Africa and western U.S.

Halophiles - Highly Salty

Natural salt lakes and manmade pools
Sometimes occurs with extreme alkalinity

Acidophiles
pH 0-1 of waters
at Iron Mountain

Alkaliphile
e.g. Mono Lake
alkaline soda lake, pH 9
salinity 8%

Halophiles
solar salterns
Owens Lake,
Great Salt Lake
coastal splash zones
Dead Sea

Survival

Temperature extremes
Every part of microbe must function at extreme
Tough enzymes for Thermophiles
Efficient enzymes for Psychrophiles

Many enzymes from these microbes are

interesting

Life at High Temperatures, Thomas M. Brock

Survival

Chemical extremes
Interior of cell is normal
Exterior protects the cell
Acidophiles and Alkaliphiles sometimes excrete protective
substances and enzymes
Acidophiles often lack cell wall
Some moderate halophiles have high concentrations of a
solute inside to avoid pickling

Some enzymes from these microbes are interesting

What are enzymes?

Definition - a protein that catalyses (speeds

up) chemical reactions without being
changed

What are enzymes?

Enzymes are specific

Lock and key analogy
Enzyme
Product B

Substrate A

Product C

What are enzymes?

Activation energy
Enzymes allow reactions with lower energy
Without Enzyme
With Enzyme

Energy

Time

What are enzymes?

Enzymes are just a protein

They can be destroyed by
Heat, acid, base

They can be inhibited by

Cold, salt

Try doing this with an egg white or milk

Protein is a major component of both

Practical Applications

Extremozymes
Enzyme from Extremophile

Industry & Medicine

What if you want an enzyme to work

In a hot factory?
Tank of cold solution?
Acidic pond?
Sewage (ammonia)?
Highly salty solution?

One solution

Pay a genetic engineer to design a super

enzymes...
Heat resistant enzymes
Survive low temperatures
Able to resist acid, alkali and/or salt

This could take years and lots of money

Extremophiles got there first

Nature has already given us the solutions to

these problems
Extremophiles have the enzymes that work in
extreme conditions

Endolithic algae from Antarctica; Hot springs in Yellowstone National Park,

1998 Reston Communications, www.reston.com/astro/extreme.html

Thermophiles

Most interesting practical applications so

far
Many industrial processes involve high heat
450C (113F) is a problem for most enzymes
First Extremophile found 30 years ago

Life at High Temperatures, Thomas M. Brock

PCR - Polymerase
Chain Reaction
Life at High Temperatures, Thomas M. Brock

Allows amplification of small sample of

DNA using high temperature process

Technique is about 10 years old

DNA fingerprints - samples from crime scene
Genetic Screening - swab from the mouth
Medical Diagnosis - a few virus particles from
blood

Thermus aquaticus or Taq

Psychrophiles

Efficient enzymes to work in the cold

Enzymes to work on foods that need to be
refrigerated
Perfumes - most dont tolerate high
temperatures
Cold-wash detergents

Algal mats on an Antarctic lake bottom,

1998 Reston Communications, www.reston.com/astro/extreme.html

Acidophiles

Enzymes used to increase

efficiency of animal feeds
enzymes help animals extract
nutrients from feed
more efficient and less
expensive

Life at High Temperatures, Thomas M. Brock

Alkaliphiles

Stonewashed pants
Alkaliphilic enzymes soften fabric and release
some of the dyes, giving worn look and feel

Detergents
Enzymes to dissolve proteins or fats
Alkaliphilic enzymes can work with detergents

Halophiles
What is a halophile?
Diversity of Halophilic Organisms
Osmoregulation
Compatible Solute Strategy
Salt-in Strategy
Interesting Facts and Applications

What is a halophile?

The word halophile means salt loving.

A halophile is an organism that can grow in higher
salt concentrations than the norm.
Based on optimal saline environments halophilic
organisms can be grouped into three categories:
extreme halophiles, moderate halophiles, and
slightly halophilic or halotolerant organisms.
Some extreme halophiles can live in solutions of
35 % salt. This is extreme compared to seawater
which is only 3% salt.

Diversity of Halophilic
Organisms
Halophiles are a broad group that can be
found in all three domains of life.
They are found in salt marshes,
subterranean salt deposits, dry soils, salted
meats, hypersaline seas, and salt
evaporation pools.

Unusual Habitats
The bacterium pseudomonas was found
living on a desert plant in the Negev Desert.
The plant secretes salt through salt glands
on its leaves.
Bacillus was found in the nasal cavities of
desert iguanas. These iguanas have salt
glands in their nasal cavities that secrete
KCl brine during osmotic stress.

Osmoregulation
Living in high salinity poses a serious stress
that halophiles have overcome through
special processes or adaptations.
The stress lies in the microbes ability to
maintain an internal osmotic potential that
equals their external environment.
Osmosis is the process in which water
moves from an area of high concentration to
an area of low concentration.

Osmoregulation
In order for cells to maintain their water
they must have an osmotic potential equal
to their external environment.
As salinity increases in the environment its
osmotic potential decreases.
If you placed a non halophilic microbe in a
solution with a high amount of dissolved
salts the cells water will move into the
solution causing the cell to plasmolyze.

Osmoregulation
Halophiles have adapted to life at high
salinity in many different ways.
One way is through the modification of
their external cell walls. They tend to have
negatively charged proteins on the outside
of their cell walls that stabilize it by binding
to positively charged sodium ions in their
external environments. If salt
concentrations decline their cell walls may
become unstable and break down.

Compatible Solute Strategy

There are two strategies that halophiles have

evolved to deal with high salt environments.
In the compatible solute strategy cells maintain
low concentrations of salt in their cytoplasm by
balancing osmotic potential with organic,
compatible solutes.
They do this by the synthesis or uptake of
compatible solutes.

Compatible Solute Strategy

Compatible solutes include polyols such as
glycerol, sugars and their derivatives, amino
acids and their derivatives, and quaternary
amines such as glycine betaine.
Energetically this is an expensive process.
Autotrophs use between 30 to 90 molecules
of ATP to synthesize one molecule of the
compatible solutes. Heterotrophs use
between 23 to 79 ATP.

 Compatible Solute Strategy

Energy is also expended in pumping out
salts that dissolve into the cell.
The uptake of available compatible solutes
in the environment is an adaptation they
have evolved to reduce the energy cost of
living in high salt concentrations.

Salt-in Strategy
Cells can have internal concentrations that
are osmotically equivalent to their external
environment.
This salt-in strategy is primarily used by
aerobic, extremely halophilic archaea and
anaerobic bacteria.
They maintain osmotically equivalent
internal concentrations by accumulating
high concentrations of potassium chloride.

Salt-in Strategy
Potassium ions enter the cell passively via a
uniport system. Sodium ions are pumped
out. Chloride enters the cell against the
membrane potential via cotransport with
sodium ions.
For every three molecules of potassium
chloride accumulated, two ATP are
hydrolyzed making this strategy more
energy efficient than the compatible
solute strategy.

Salt-in Strategy

To use this strategy all enzymes and

structural cell components must be adapted
to high salt concentrations to ensure proper
cell function.

Halobacterium: an example of an
extreme halophile
Halobacterium are members of the halophile
group in the domain archaea. They are
widely researched for their extreme
halophilism and unique structure.
They require salt concentrations between
15% to 35% sodium chloride to live.
They use the salt-in strategy.
They produce ATP by respiration or by
bacteriorhodopsin.

Halobacterium
They may also have halorhodopsin that
pumps chloride into the cell instead of
pumping protons out.
The Red Sea was named after
halobacterium that turns the water red
during massive blooms.

Facts
The term red herring comes from the foul
smell of salted meats that were spoiled by
halobacterium.
There have been considerable problems
with halophiles colonizing leather during
the salt curing process.

Applications

Current applications using halophiles

include:
the extraction of carotene from carotene rich
halobacteria and halophilic algae that can then
be used as food additives or as food-coloring
agents.
the use of halophilic organisms in the
fermentation of soy sauce and Thai fish sauce.

Applications

Many possible applications using

halophiles are being explored such as:
increasing crude oil extraction
genetically engineering halophilic enzyme
encoding DNA into crops to allow for salt
tolerance
treatment of waste water

Conclusions
Halophiles are salt tolerant organisms.
They are widespread and found in all three
domains.
The salt-in strategy uses less energy but
requires intracellular adaptations. Only a
few prokaryotes use it.
All other halophiles use the compatible
solute strategy that is energy expensive but
does not require special adaptations.

Genetic prospecting

What is it?
Think of a hunt for the genetic gold

Pr. Patrick Forterre, Extremophiles Laboratory of IGM at Orsay, France http://www-archbac.u-psud.fr/

Summary

Now you know something about

Extremophiles
Where they live & how they survive

They are interesting because

They have enzymes that work in unusual
conditions
The practical applications are interesting