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[Music]

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Thank You Moran for the nice

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introduction so before I jump into my

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solutions for the phosphorylation

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problems I thought I would just give a

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[Music]

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give a little bit of the core philosophy

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and the core view that we have in the

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center for chemical evolution so while

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the RNA world looks really good and

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probably existed at some time we don't

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believe that that was the very first

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form of life that existed on this planet

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we believe that there was something that

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preceded it and we use clues from RNA

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and DNA to figure out what could have

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preceded that and so one of the ways

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that we do that is we look at modern RNA

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in DNA and break it down into its three

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central components you have the

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recognition units the basis that it uses

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to store the information and also what

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can do the base pairing and solution you

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have the ionized linker something that

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has some sort of charge on it modern

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life uses phosphate and this helps it to

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soluble eyes in solution and also help

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the base pairing to happen by repelling

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the negative charges and then you have a

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tri functional connector a bridging

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molecule that keeps these pieces

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together and has nice chemical

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functionality to help chemistry and

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biochemistry down the line so in our

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Center we work on changing out these

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pieces from RNA and DNA and putting

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other pieces in there and seeing how

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this affects its properties and if you

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can have life without having the

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specific backbone as we see in modern

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biochemistry so we've had a good deal of

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success replacing the bases for the

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recognition units and replacing the Tri

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functional connectors but we've run into

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some problems when we tried to change

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that ionized linker into other moieties

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with our group and other groups in the

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world it just doesn't work as well so as

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opposed to trying to change it where

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some of us in the lab are trying to

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embrace it what if we had phosphate at

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the origins and it got incorporated into

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the molecules but this is fraught with

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some problems that have haunted the

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astrobiology community for the past 50

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to 60 years so one of the first issues

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is how do you even react it with early

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molecules how do you get phosphate

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incorporated in so these reactions are

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not favored in water at all it drives

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off water it's very hard to do in a

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water-based reaction and so modern

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biochemists use organic solvents and dry

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conditions to actually add the

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phosphorylation which is problematic

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because you can view the early Earth

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just covered in water and so it's nice

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to find some water or liquid based

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chemistry to make this happen and

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another major problem is the phosphate

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availability issue so a lot of the

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phosphate that we see on modern earth

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and we see from the mineral records is

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bounded insoluble species with the

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divalent cations and so this is

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traditionally been considered to be

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extremely inaccessible for prebiotic

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chemistry so how do we knock out these

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problems one at a time so how did the

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phosphorylation chemistry even happen so

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a remarkable molecule that's been

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studied for the past 40 years is urea

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this is actually the first organically

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synthesized molecule from waller in the

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early 19th century and it's a very it is

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thought to be extremely abundant on a

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prebiotic earth

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and one of the things that we all like

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to describe is you have the salt flats

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that you can see throughout the world

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now like this one depicted in Utah well

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on a prebiotic

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earth urea would have been so abundant

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that we probably would have had your

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rear flats just like we have salt flats

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so this would be very abundant and found

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everywhere and when you take a very

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simple reaction you mix in some soluble

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phosphate with urea and different sugars

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you have no water and just add heat a

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hundred degrees or more you actually

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form the phosphorylated product so this

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has been discovered about 40 years ago

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and it's a very nice reaction and a good

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way to get phosphate attached but it

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requires high temperatures that degrade

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a lot of the molecules that you want to

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phosphorylate and creates an awful lot

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of side products it's a very messy

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reaction with low yields but it does

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work so we have some chemistry that can

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work well how do you solve the problem

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of running it in the presence of water

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so as I pointed out you can see the

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reaction at the bottom just attaching

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phosphate to a simple molecule like

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glycerol it drives off water and you

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form the bus for the the phosphorylated

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organic down there it doesn't work if

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there's water because you need to remove

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the water so one of the things that

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we've started to look at based upon some

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previous work is using eutectics

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and eutectics are very unique systems

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you can take two solids like is depicted

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on the left and the right there

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mix them together and they actually form

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a liquid so it lowers the melting point

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of both of them you can take it to very

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low temperatures in a water free

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environment and so the tactics that I

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use for my experiments they do start

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with excess water but we heat them open

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to the atmosphere and drive off most of

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the excess water and create this nice

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liquid environment

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that goes down to very low temperatures

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so now we have this now we have this

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eutectic which I use urea to make

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so it's a very urea rich environment

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that's liquid and you can do a lot of

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very interesting fluid based chemistry

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at low temperatures and the tactics that

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I use are made of very valuable

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components of prebiotic ly so I have a

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ammonium formate and ammonium acetate we

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look at different chemical synthetic

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reactions that would have been likely on

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a prebiotic earth and see a lot of these

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just pop out and one of the wonderful

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things if you mix these together it

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evaporates off most of the compounds and

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you get a very specific ratio in a

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perfect ratio for doing a lot of the

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chemistry that we're looking for so you

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can start with any concentration that

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you want it all converges upon what we

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need so we've found a reaction medium

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that removes the water from it so that's

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cool we have two steps down already but

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where do we get the phosphate from and

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this has traditionally been the biggest

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problem with prebiotic phosphorylation

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so most of the minerals would be present

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in or most of the phosphate would be

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bound in a hydroxyl apatite which is

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extremely insoluble so it's bad for

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doing chemistry but it's good for modern

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life as this is what we make up when we

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used to make up bounce and if bones are

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that's a problem these don't dissolve

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you don't have any phosphate in your

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solution so we start looking at the

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modern earth as an analogue and try to

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figure out where we could have got

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phosphate from and see if it would be

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similar to what we could have on a

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prebiotic earth so what are the

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environments depict at the top is laguna

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santa maria in argentina and we see that

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this is a phosphate rich system that is

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in this aqueous pool so that seems great

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but you take a closer look and you see

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that the phosphate is coming from all of

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the biomass that's in there so it's not

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really a

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source it's just a biological source but

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then one of my collaborators remembered

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some of his work that he did in Spain

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just looking at of all things pig urine

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and waste pops so these are urea

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magnesium and calcium rich environments

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that just form giant waste pools and

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when you look at them you would expect

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to see the calcium phosphate the

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hydroxyl apatite babe depicted before

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but instead you see this mineral called

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struvite which is a much more soluble

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form and you see that to the exclusion

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of any other minerals in there so

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instead of form a hydroxyl apatite it

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gets trapped in this more soluble form

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so we started thinking can you do this

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in a laboratory so we tried to

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synthesize the struvite which would be a

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much better source for phosphorylation

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and we thought about this in terms of a

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prebiotic model so you can have a pool

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on the early earth full of calcium

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magnesium and phosphate it would

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precipitate and form this mineral at the

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bottom then you could drive off all the

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water you have some local outgassing

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from volcanoes and other hydrothermal

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vents and it could form it could provide

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magnesium and sulfate and urea and

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formate the way you come down wash it

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into the pool and then you get the

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conversion to the exact form of minerals

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that we need down there so instead of

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calcium phosphate now you have a

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struvite layer at the bottom and you've

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done it you've mobilized the phosphate

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so we tested this in the laboratory we

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simply took the hydroxyl apatite mix it

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with the eutectic heated it for seven

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days with adding an magnesium sulfate

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and lo and behold we saw the conversion

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of hydroxyl apatite disturb it-- and we

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have the XRD and the rom inspector over

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there to help to prove it but we saw

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basically quantitative conversion in

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these conditions which is fantastic so

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we wanted to put it all together and see

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if not only you could convert but if you

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could also phosphorylate so one of the

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things we looked at for prebiotic

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chemistry is you want to keep it simple

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the early Earth didn't have a lot of

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tools and neither should we so I just

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run my reactions on a simple plate by

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mixing everything together so I take

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adenosine as my model molecule mix it

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with any phosphate source mineral or

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soluble phosphate heat it from fifty

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eighty five degrees and they do get

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phosphorylated identity and one of the

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nice things I put up this chromatogram

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up here just to show how clean these

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reactions are so every peak on there

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represents a different species and so I

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get all of these phosphorylated species

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and not many other side reactions so

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this is a lot cleaner than the previous

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work and overall I've created for

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different eutectics that I show up here

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the part that outlined in red is using

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soluble phosphate and you can see I can

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get upwards of 90 percent

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phosphorylation when phosphate but when

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the phosphate is in water or in the

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eutectic soluble when I take the

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struvite or Newberry as I have depicted

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here which is moderately soluble the

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phosphorylation does go down but I can

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still get better than 20% so that is a

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pretty good mineral for phosphorylating

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you can't expect 100% yields from

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everything but one of the most

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remarkable things is with the

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hydroxyapatite down there we get a small

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amount of phosphorylation which is

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better than what is shown here with just

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the urea where you get none and even

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more interestingly when we add magnesium

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to the reaction magnesium sulfate

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a doubling or tripling of the amount of

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phosphate so we take these eutectics

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just add some magnesium sulfate and we

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get a good amount of phosphorylation

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helping to address a long-held problem

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in prebiotic chemistry so that overall

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conclusions the phosphorylation is

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robust it happens in a wide range of

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mixture it's not a very specialized

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reaction it's absorbed at moderately low

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temperatures as low as 50 degrees which

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is very easy on prebiotic earth it's

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successful with the insoluble hydroxyl

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apatite and it's significantly improved

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in the presence of magnesium sulfate

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these are very prebiotic viable

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conditions and so I'd like to

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acknowledge my collaborators from many

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different Institute's from the

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University of southern Florida and from

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the University of the University da de

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alcalá

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and my group and my funding sources and

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I'd like to take any questions now all

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right catch it's worked all right is the

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is the mechanism there more tied to the

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heating or to the drying the mechanism

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is tied to the urea in the absence of

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water and so you do need some heat there

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in order to actually activate the

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phosphate okay so it's not about driving

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though it's not about driving the water

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off it's about it's about both okay okay

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if I try to run it in water if I close

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the vials it doesn't work now yeah what

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about the pH for example we have

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hydroxyapatite which is not soluble but

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the Fiat sulfuric acid which is common

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and we'll connect settings you just

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leash all the phosphate immediately so

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it's not a problem on the more acidic

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page in 7 sure yeah and so that's one of

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the things

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can have extremely low PHS which will

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sell utilize it and that's fine but it's

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hard to preserve a lot of the chemicals

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that you need and do other reactions in

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an extremely acidic environment so you

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can view it as a multi-step problem

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where you dissolve it you possibly

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phosphorylate it and then you move it on

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to another system so that's been the

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traditional view of it before but it's

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highly contentious because of the other