Lab notebook quarter 1

Homework: Find the largest observed gradient in biology (may be something like a plant
in low salt setting), assume the vacuole is the entire cell, what amount of yeast could we
use to desalinate water assuming we can get this gradient? How many runs would it
take? Put your calculation in your lab notebook.
http://www.jbc.org/content/272/42/26145.full
Describes sequestration of intracellular NaCl
Cerevisiae doesn't have a specific salt-uptake system and Na+ enters through a K+ transport
where K+ is preferred over Na+ http://aem.asm.org/content/63/10/4005.full.pdf+html
http://link.springer.com/article/10.1007/BF00164757#page-1
Metal uptake up to three times greater with glucose. Protective qualities
Denatured biomass not that much less effective at salt intake !
http://onlinelibrary.wiley.com/doi/10.1046/j.1365-313X.2003.01958.x/full
Plant storing up to .6M (seawater) in vacuole
http://onlinelibrary.wiley.com/doi/10.1002/yea.320060303/abstract
D hansenii: at 25C, pH 50 were the following: Vmax, decreasing from over 500 to less than 40
mol g

per h over a concentration range of 03 M extracellular sodium chloride; Km

(glycerol) 04006 mM over the same range; Km (sodium ions) 160 321M; Km, (potassium
ions) 104 36M.
Osmotic pressure (pi) = (mol solute) * (ideal gas law) * (temperature) / (volume)
Questions
What is the max salt intake rate?

For D Hansenii it is 500 mol g

per h in ideal conditions, in seawater would be less
What is the max salt storage assuming cell is whole vacuole?
What if salt was biosynthesized into other products? What could that be?
How do we make sure yeast wont immediately die?
May die immediately if the osmotic pressure is too great. Maybe survive some time even
if salt concentration is too great.
Why does denatured biomass still uptake about the same?
Explain ion transport systems and gradients. Is membrane potential important (voltage)?
How come metal cations are taken up so much faster with presence of glucose?
How difficult could it be to make a salt uptake system for cerevisiae? Only has a K+ transport
system
integrate foreign salt transport systems
How do I begin searching for the "largest gradient"

Needs further research

Glycerol sodium pumps D hansenii, potassium sodium pumps, general info on membrane
potential and ion gradients
Calculations for volume of yeast needed for desalination for given salt gradient
Seawater = .6 M concentration = .6 moles/L
Lets say max observed gradient we want to replicate is X molar of NaCl in H2O and Y molar of
NaCl in yeast
Solution volume = yeast volume + water volume
amount of salt in water + amount of salt in yeast = .6 moles
X(water volume) + Y(yeast volume) = .6 moles
X(solution volume - yeast volume) + Y(yeast volume) = .6 moles
X(solution volume) - X(yeast volume) + Y(yeast volume) = .6 moles
yeast volume = ( .6 - X(solution volume) ) / (Y-X)
Example: Lets say our yeast can store 1 M of NaCl and the water outside the cell is .1 M of
NaCl in 1 L of water.
(1 M * yeast volume) + (.1 M * water volume) = .6 mol
(1 M * yeast volume) + (.1 M * (1L - yeast volume) ) = .6 mol
(1 M * yeast volume) + (.1 M * 1L) - (.1 M * yeast volume) = .6 mol
.9 M * yeast volume = .5 mol
yeast volume = .5/.9 L
This is a 83.3% decrease in volume when 55.6% of the solution is yeast.
Ideas for Na+ or Cl- sequestration
https://en.wikipedia.org/wiki/Ethylenediaminetetraacetic_acid
common food additive. Sequesters cations. Soluble and tends not to form precipitates
https://en.wikipedia.org/wiki/15-Crown-5
Crown ether form complex with Na+