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!! Introduction to Extremophiles 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
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