BIO 305 Exam 2 Chp 4-6 Flashcards
What are the physical requirements for growth?
Temperature, pH, and osmotic pressure.
What are the chemical requirements for growth?
Carbon, nitrogen, oxygen, hydrogen, sulfur, phosphorous, and other trace elements. (Most of the macronutrients).
What are essential nutrients?
Essential nutrients=nutrients required for the growth that the organism cannot make on its own.
What are macronutrients?
Macronutrients:
Carbon, nitrogen, hydrogen, oxygen, phosphorous, and sulfur.
Used a lot to make macromolecules.
What are cofactors?
Cofactors are other important macronutrients but they are not used for macromolecules.
Mg2+, Fe2+, and K+ are important for enzyme function.
Ca2+ is a cofactor that acts as a secondary messenger.
What are micronutrients?
Micronutrients=nutrients required for growth but only needed in small amounts.
Ex: Cobalt, zinc, and copper manganese.
What are growth factors?
Growth factors=factors needed for some organisms to grow but not for all of them.
They may have evolved due to the organism’s environment and what it provides.
What is defined minimal media?
Defined minimal medium=contains the minimum requirements for organisms to grow.
What are obligate intracellular parasites?
Obligate intracellular parasites=those that are impossible to grow outside of the host.
What is axenic growth?
Axenic growth=growth of a single organism in culture.
How has metagenomics helped discover new organisms?
It shows new genetic material that does not match any known organisms. It shows uncultured organisms.
What does LB Broth grow?
Gram-negative and Gram-positive cells.
What does MP Medium grow?
Gram-negative cells like E. coli.
What is proton motive force?
Proton motive force=moving of H+ across the membrane.
How is the proton motive force useful in generation ATP?
It is the force and that causes protein changes and energy provider to anabolic build ATP.
What are CFUs?
CFUs=colony forming units.
While the assumption is that one cell=one colony, is that always true?
No. Some bacteria, like Staphylococcus, are bunched together so multiple cells form a colony.
What are the important properties of agar that make it so useful?
Microbes can’t degrade it.
Liquefies at 100C.
Solidifies at 40C.
Once solidified, it will not become liquid until 100C again.
What is the difference between streak plating and spread plating?
While both can use dilution, streak plating paints bacteria onto an agar plate so there are fewer bacteria with later streaks. Spreading plating lays bacteria on an agar plate by using liquid.
What is sporulation?
Sporulation=formation of spores.
What is germination?
Germination=return of a cell to a vegetative (growing) state.
What does the suffix “-trophy” mean?
Trophy=acquistion of nutrients.
What is a chemolitoautotroph?
Carbon source: reducing CO2.
Energy source: chemical reactions.
Electron source: Inorganic molecules.
Uses inorganic molecules for e-. Energy from oxidation is used to reduce CO2 for a carbon source.
What is a photolithoautotroph?
Carbon source: reducing CO2
Energy source: Light.
Electron source: Inorganic molecules.
To capture energy from light, photolysis of inorganic molecules is used to excite their e-. The energy from capturing light is used to reduce CO2 for a carbon source.
What is a chemoorganotroph?
Carbon source: Digesting organic molecules.
Energy: Chemical reactions.
Electron source: Organic molecules.
Gain carbon and e- from an organic source. The e- from the organic source provides energy through chemical reactions.
What is a photoorganotroph?
Carbon source: Digesting organic molecules.
Energy: Light.
Electron source: Organic molecules.
Organic compounds are brought into the cell. Energy is gained from their breakdown through electrons and the absorption of light. Organic compounds are used to build biomass.
What do nitrogen-fixing bacteria do? What do they gain from this process?
Turn N2 into NH4 (ammonium). Nitrogen can now be integrated into organic molecules.
What do nitrifiers do? What do they gain from this process?
Nitrification=conversion of ammonia, or ammonium to nitrate (NO3-).
This is lithotrophy and they use ammonia and ammonium for electrons.
What do denitrifiers do? What do they gain from this process?
Denitrification=NO3- to N2.
Reduce NO3- to N2 by using it as the final electron acceptor.
What is liquid media useful for?
Liquid media is useful for observing growth kinetics.
What is solid media useful for?
Solid media useful for isolating a single species.
What is complex media?
Complex media=components not defined, rich in nutrients.
What does complex media not tell us about the organisms growing in it?
Rich media does not allow us to know exactly what the organism needs to grow.
Since we don’t know exactly what is in it, we don’t know what growth factor they are relying on.
What is synthetic media? Do all organisms grow in it?
Synthetic media=components defined. No, because there may be missing growth factors.
What is defined media?
Defined media=media with only essential nutrients.
Can be derived from synthetic media.
What is enriched media?
Enriched media=compoents of blood are added to media.
These are for fastidious organisms.
What is selective media?
Selective media selects for one organism over another.
What is mannitol salt agar? What kind of media is it?
Selective media.
Mannitol salt agar has high salt conditions.
Ex: Promotes growth of Staphylococcus.
What is Sabouard’s dextrose agar? What kind of media is it?
Selective media.
Sabouard’s dextrose agar has a pH of 5.6 and is used to isolate fungi.
Used to grow yeast.
What is brilliant green agar? What kind of media is it?
Brilliant green agar=inhibits gram-positive and most gram-negative intestinal bacteria except for Salmonella.
What is differential media?
Differential media=selects for multiple organisms.
What is MacConkey agar? What kind of media is it?
Differentiating media.
Uses lactose as the differentiating part.
Uses bile salts to select for Gram-negative bacteria.
Gram-positive bacteria are less resistant to bile salts.
Distinguishes by color due to metabolism of different sugars causing a pH change.
There is a pH indicator in the media that shows a color difference among the different species.
Explain the Petroff-Hausser method of counting bacteria. Pros and cons?
Uses hemocytometer to count cells.
Very simple.
The count can be skewed by dead cells that are not distinguished from live cells.
Explain using fluorescent stains to count bacteria. Pros and cons?
Stains cells based on whether they are alive or dead. Ex: Propidium iodide, a red dye, intercalates in DNA but cannot penetrate the membrane of live cells.
Can distinguish between live and dead cells.
Equipment is expensive.
Explain the flow cytometry method for counting bacteria. Pros and cons?
Sorts cells based on expression levels of fluorescence which can be indicative of protein expression levels.
Can sort high expressing cells from low expressing cells.
Equipment is expensive.
Explain the viable count method for counting bacteria. Pros and cons?
Uses dilutions of bacteria on a streak or spread plate to find a number of CFUs.
Counts only living cells.
Takes time.
Not every CFU comes from one cell.
May underestimate the number of living cells. Damaged cells will be alive but cannot divide.
Explain the biomass method of counting bacteria. Pros and cons?
Dries out cells and weighs them.
Measures the overall size of a bacterial population.
Time-consuming.
Need a lot of mass because cells are so small.
Explain the biochemical assay method of counting bacteria. Pros and cons?
Measures protein content and metabolic rate.
More sensitive.
More accurate than biomass.
Some biochemical pathways fluctuate so the number will not always be the same and representative of how many cells there actually are.
What is the optical density method for counting bacteria. Pros and cons?
Uses spectrophotometer to measure how much light is absorbed by a dilution of culture. Quick assay. Cells can still get stuck together. Constants are not perfect. Only provides an approximate result. Dead and live organisms scatter light.
Explain the PCR method for counting bacteria. Pros and cons?
Show quantification of DNA.
Doesn’t take too long to get results.
Dead and live cells have DNA.
What is the lag phase of growth?
Lag phase=little growth as bacteria get adjusted to the new environment and nutrient availability.
A lot of protein synthesis in this step.
What is the exponential phase of growth?
Exponential growth=great growth rate as bacteria grow and use up nutrients.
Bacteria are the most vulnerable here.
What is the stationary phase of growth?
Stationary phase=equal number of cells dying and growth.
Growth slows due to fewer nutrients.
Bacteria are more resistant here.
What is the death phase of growth?
Death phase=deaths outnumber growth.
If media is not replaced, this will occur.
What is generation time/doubling time?
Time it takes for a cell to divide.
What are the steps for endospore formation?
Septum does not form in the middle of the cell, but closer to one of the poles.
Septum forms and forespore is created.
Replicated DNA is placed into the forespore.
Mother cell engulfs forespore.
The chromosome of mother cells disintegrates.
Exosporangium and protein cover are generated around the forespore for extra protection.
A layer of peptidoglycan is built between the forespore membrane and membrane made by the mother cell.
Spore coat includes calcium and dipicolinic acid is used to protect DNA.
Spore is released into the world.
What is the biofilm life cycle?
Some bacteria adhere to a surface.
The attached cells make a microcolony.
Biofilm gets larger and an exopolysaccharide (EPS) layer is formed for structure and protection.
A mature biofilm is generated.
Dissolution of biofilm occurs when nutrients become scarce.
What are the important aspects of a biofilm?
Exopolysaccharide layer=holds biofilm together.
Increased tolerance and resistance helps bacteria to survive.
Cell differentiation.
Different jobs based on where the cell is and what nutrients they have access to.
Cells that attach to a surface.
To keep biofilm attached to the surface.
Is the foundation for the rest of the biofilm.
Quorum sensing for communication.
Bacteria sense individually and do a respond as a group.
If bacteria can’t do quorum sensing, they can’t find a biofilm.
Explain the differentiation of cyanobacteria between oxygen-producing and nitrogen-producing bacteria.
For cyanobacteria species, (ex: Anabaena) every 10th cell becomes a heterocyst that fixes nitrogen.
The cyanobacteria must differentiate this job because the enzyme nitrogenase, which fixes bacteria, is very susceptible to oxygen and ROS.
Heterocysts have 3 extra cell walls and a special barrier to keep out O2.
What do fruiting bodies do? Explain their differentiation.
Fruiting bodies (such as in Myxococcus xanthus), form a biofilm under nutrient stress.
They are able to glide using pili.
Cells on the interior make spores that then spit out the biofilm.
The hope is that they will find a spot with better nutrients.
Explain the differentiation and life cycle of filamentous biofilms.
Filamentous biofilms can be formed with Anticomycetes species that generate filamentous hyphae that form mycelia. (This is similar to fungi).
Once a foundation in the soil is formed, aerial mycelia can be formed.
In high glucose time, exploratory cells will be sent to find the nutrient-rich areas.
In low glucose areas, aerial hyphae cells will be produced.
bld gene causes the production of aerial hyphae.
Once the growth stops, the spores of the aerial mycelial form.
The compartment segments of the aerial hyphae are formed from the whi gene.
How does ATP synthase work?
Uses protein motive force where H+ goes down its concentration gradient into the cell.
Proton motive force causes protein motor to spin and protein structural changes which allow for the construction of ATP.
This would not happen if energy were not provided for the change.
What is a symport?
Symport=molecules move in the same direction.
What is an antiport?
Antiport=molecuels move in different directions.
What is coupled transport?
A molecule moving down its concentration gradient provides energy to transport a molecule going up against its concentration gradient.
What is an ABC transporter?
ABC transporters=ATP-binding cassette superfamily.
Uses ATP for active transport.
What is uptake active transport?
Uptake ABC transporters move molecules into the cell against their concentration gradient.
Uses a cognate to match a solute with a solute-binding protein to bring solutes to the ABC transporter.
What does cognate mean?
Cognate=matched.
Ex: Siderophores that have a high affinity for iron and bring it to an ABC transporter.
What is efflux active transport?
Efflux ABC transporters move molecules out of the cell against their concentration gradient.
How is ATP used with ABC transporter?
ATP binds to ATP-binding cassette proteins on the ABC transporter.
Hydrolysis of ATP provides energy from uptake or influx.
How does group translocation work?
By chemically changing a product, you make it different so it can go down its concentration gradient.
It doesn’t match the normal solute on the outside and makes its own gradient with itself.
Ex: Phototransferase system with glucose to add a phosphate to it.
What is considered a normal environment? Temperature, pH, and osmolarity?
20-40C
pH=7.0
0.9% salinity.
What are extremophiles?
Extremeophiles=microbes that live outside of normal environmental ranges.
Does temperature increase always increase metabolism?
It does to a certain point because higher temperature means more molecular motion. However, for each cell’s maximum temperature, enzymes and proteins will start to denature because it is too hot. When that happens, production goes down and the cell dies.
What are some reasons that cells might not want their proteins to all have the same temperature range?
The same enzymes are used in stress response.
Some only need to work in cold or hot temperatures.
Makes microbes more susceptible if everything has the same temperature range.
Not every protein has the same end goal.
Cells don’t want to grow as rapidly as possible because they would suck all their nutrients.
What are barophiles?
Barophiles (or piezophiles) can live at the bottom of the ocean where pressures reach 400-1,000 atm.
These bacteria are usually psychrophilic or thermophilic (if next to a heat vent).
Require intense pressure to survive.
What is water activity (a(w))?
Water activity (a(w))=availability of water.
What is osmolarity?
Osmolarity=concentration of solutes in solution.
What relationship do water activity and osmolarity have?
a(w) and osmolarity have an inverse relationship.
What are compatible solutes?
Compatible solutes=solutes that the cell can have a lot of with no negative effects.
What are mechanosensitive channels?
Mechanosensitive channels can release solutes when the pressure becomes too high.
What is plasmolyzing?
Plasmolyse=cell membrane pulls away from the cell wall and the cell lyses.
What are halophiles?
Halophiles require high salt.
2-4 M NaCl.
What are obligate halophiles?
Obligate halophile=require 30% salt solution to grow.
What are facultative halophiles (halotolerant)?
Facultative halophiles=can grow in high salt conditions but do not require it.
Can grow from 2%-15% salt.
What are strict aerobes?
Strict aerobes=need O2 to survive.
What are strict anaerobes?
Strict anaerobes cannot use O2 and do not have enzymes to remove ROS.
What are facultative aerobes?
Facultative microbes=can survive with and without oxygen.
E. coli and yeast.
What are microaerophiles?
Microaerophilic=can withstand a small amount of O2.
What are aerotolerant microbes?
Aerotolerant=cannot use oxygen but can tolerate it well.
Have enzymes to destroy ROS.
What makes oxygen so dangerous and toxic?
Makes free radicals that can harm cells by oxidizing reduced products.
Part of the aging process.
What is oxygen used for in aerobic respiration?
It is used as the final electron acceptor.
What does superoxide dismutase do?
Superoxide dismutase=ROS enzyme that turns O2- to H2O2.
What does catalase do?
Catalase=turns 2H2O2 to 2H2O and O2.
What does peroxidase do?
Peroxidase=turns 2H2O2 to 2H2O and NAD+.
What are reactive oxygen species?
ROS=H2O2, Superoxide (O2-), and hydroxyl radical.
Happens when oxygen is not completely reduced.
Mistakes are more likely to occur as organisms age.