Aspartic Acid in the aquarium

[Habib]

Randy:

I don't expect an answer to the questions

Thank you that saves a lot of time
Just kidding.

The questions you have are very logical and have therefore been investigated or have been addressed.

I will give you ASAP answers and opinions and try do discuss them.

 

[saltshop]

Interesting thread lot's of questions.....

Aspartic Acid = NutraSweet(tm)?? or at least a large portion of it??

Carboxyl groupings do the binding to Calcium?

How is aspartic acid gained by the coral, diffusion from the water or through food stuffs?

Is aspartic acid the most ideal organic acid or is it simply the most abundant? I am aware that both humic acid and fulvic acid can be found in high densities in the organic matrix from periods of high rain and terrestrial run-off. They are used to determine rainfall and run-off dating back in time through examination of coral skeletons. Do humic anf fulvic acid replace aspartic acid during these periods? If so, why?

What is the level of aspartic acid in both seawater and our tanks? What is the level of other organic acids as well?

If the level is low in our tanks, given that they are usually a pretty rich organic soup, do other organic acids take their place? Is it actually limiting?

Since corals throw just about everything but the kitchen sink into their skeletal matrix again I wonder if it is simply an issue of availability in sea water. In years of volcanic erruptions you find a large increase of heavy metals in skeletons and even radioactive elements such as Radium and Barium are included in the matrix at much higher levels than what is found in NSW. Does the appearance of an element, or organic in this case=requirement or other?

I have little doubt that the organic portion of the matrix is needed, but would really like to see a lot more before a conclusion could be made that says "yes, aspartic acid is preferred and needed over other possible organics that may be being produced in our tanks". I realize no one here is making any conclusions and I too think it would be interesting to see if the addition of certain organics makes any sort of difference in our tank, am just supplying food for thought.

 

[Habib]

JB,

I expect to post, with respect to Randy's questions, more details, opinions and answers within a few days.

But allow me to respond your post.

Aspartic Acid = NutraSweet(tm)??

At least here in Holland we don't know NutraSweet. But if it is an artificial sweetner then it is most probably L-aspartyl-L-phenylalanine methylester. Thus not aspartic acid.

Carboxyl groupings do the binding to Calcium?

Many, if not all, weak organic acids will be attracted to a calciumcarbonate surface. Same priniciple for (organic)phosphate(s) and perhaps also silicic acid (currently being investigated by us)

How is aspartic acid gained by the coral, diffusion from the water or through food stuffs?

Wait till I have replied to Randy's questions. But in short experiments with labelled aspartic acid have shown that it can be taken up from the water and transported exactly to where it is needed .
Note that I say "can be taken up" , I am not ruling out that all requirements are met by e.g. food.

Is aspartic acid the most ideal organic acid or is it simply the most abundant?

See my upcoming reply.

I am aware that both humic acid and fulvic acid can be found in high densities in the organic matrix from periods of high rain and terrestrial run-off. They are used to determine rainfall and run-off dating back in time through examination of coral skeletons. Do humic anf fulvic acid replace aspartic acid during these periods?

I don't know if they actually replace aspartic acid or are adsorbed on sites which were at that moment free of aspartic acid.

If so, why?

Depends on "respective strengths of associations" , concentration ratios (e.g. aspartic acid / humic acid) , kinetics, presence of other ions (e.g. iron binds very strongly to humic acids consequently neutralizing humic acids), ...............

What is the level of aspartic acid in both seawater and our tanks? What is the level of other organic acids as well?

If we would know it then it would certainly be a concentration range. Furthermore if standing stock concentrations are known then they do not give information about input and output.

I believe that in our booklet describing our product which contains a.o. aspartic acid we say ....our aquariums lack the bio diversity of a reef.......some essential amino acids, amino sugars,....could be in a too low concentration...
At the same time we also explain that organic layers are present etc. etc.. So at the time we designed that product (8 years ago?) we ourself had questions and still do suchas which organisms excrete aspartic acid, in what amount and why,......

If the level is low in our tanks, given that they are usually a pretty rich organic soup, do other organic acids take their place? Is it actually limiting?

I think that if you look at what I have said a.o. regarding humic acids then same would count for organic acids in general.


Since corals throw just about everything but the kitchen sink into their skeletal matrix again I wonder if it is simply an issue of availability in sea water.

So what is the difference with for instance humans when we not only consider our skeletal material but also other tissues?

OK we can some times walk away or decide to move to a less polluted area or refrain from smoking or eating certain products.
But if we were not able to do so?

And does it mean that all we have accumulated or have in our body when lived in a clean environment etc. has no function?

I have little doubt that the organic portion of the matrix is needed, but would really like to see a lot more before a conclusion could be made that says "yes, aspartic acid is preferred and needed over other possible organics that may be being produced in our tanks".

IMO there are already some clues in the data I have given in the above posts.

I realize no one here is making any conclusions

Not publicly so far but is the intention and takes time.

am just supplying food for thought

Thanks, JB.

 

[saltshop]

Thanks for the thorough reply Habib! I am looking forward to the rest of what you have to say as well. Is it possible for you to say what product it is that you are talking about? I think we all realize that you are not here to try to sell your products (heck they are good enough they pretty much sell themselves) so I don't think it would be a problem if you listed it. 8 years working on this...wow! Thanks again....

 

[Randy Holmes-Farley]

Saltshop:

To add to Habib's comments:

Carboxyl groupings do the binding to Calcium?

Yes, and aspartic acid would be better than almost all other amino acids because it has two carboxylic acid groups. It is :

HO2CCH2CH(NH2)CO2H

I wish I could upload a structure, but this will have to do. At pH 8.2, it will be in these two forms:

-O2CCH2CH(NH2)CO2-

and

-O2CCH2CH(NH3+)CO2-

Both have two negative carboxylates to bind Ca++.The top form might even be able to chelate a single Ca++ between the two carboxylates (I'm not sure on this). If so, that would boost the binding tremendously.


How is aspartic acid gained by the coral, diffusion from the water or through food stuffs?

I don't have any data on this, but it sounds like Habib does.

Is aspartic acid the most ideal organic acid or is it simply the most abundant?

Aspartic acid is an amino acid that the corals need for calcification. Presumably they make a protein or peptide out of it. Humic and other acids may bind strongly to CaCO3, but that doesn't mean that a coral can use them for whatever purposes it has in mind for the aspartic acid-containing protein.

What is the level of aspartic acid in both seawater and our tanks? What is the level of other organic acids as well?

Good question.

If the level is low in our tanks, given that they are usually a pretty rich organic soup, do other organic acids take their place? Is it actually limiting?

Another good question, but one I feel comfortable speculating on the answer. If any organism makes a specific protien for a specific use using specific amino acids, it would be highly unusual to be able to substitute other amino acids if one were in short supply. Not unheard of, certainly, but very, very rare.

Does the appearance of an element, or organic in this case=requirement or other?

No clearly not. But aspartic acid is apparently needed for calcification, and it does not apparently come from the coral itself. Does that mean that we need to add more than is in the tank naturally? Again, certainly no. But it leaves open the possibility that it MAY be in short supply, and there may be a benefit from adding it.

am just supplying food for thought.

I agree with Habib: thanks. This type of discussion is what makes the hobby, and this forum, most interesting to me.

 

[saltshop]

Hi Randy-

Thanks and your welcome. I will sit down, be quiet, and wait for Habib...must be a long post to come if it is going to take a few days.

I wish I could upload a structure, but this will have to do

Will this work has both 2D and 3D?
http://www.math.smith.edu/~amino/aspartic/aspartic.html

As a side note-interesting reading on aspartame (broken down during digestion into aspartic acid, hence the confusion from above when NutraSweet was brought up). There really are more wackos out there than I thought, but it was entertaining to say the least.

 

[Randy Holmes-Farley]

Will this work has both 2D and 3D?

Yes, that's perfect. That should help people to understand the structure who can't make out the structure from what I was writing.

There really are more wackos out there than I thought, but it was entertaining to say the least.

What kinds of things were they saying?

 

[Habib]

JB:
...must be a long post to come if it is going to take a few days.

I am always busy with 10 different things, English is not my native language, use too much artificial sweetner so that I can't concentrate...

I will sit down, be quiet, and wait for Habib...

OK just in case you have difficulty in being quite...

Here is one to keep you quite for a few minutes:

Dynamics of calcification in the mediterranean red coral, Corallium rubrum (Linnaeus) (Cnidaria, Octocorallia)
Denis Allemand *, Sylvie Bénazet-Tambutté
Observatoire Océanologique Européen, Centre Scientifique de Monaco, MC-98000 Monaco, Principality of Monaco


Abstract
The spatial and temporal dynamics of calcification in whole colonies of Mediterranean red coral Corallium rubrum (Linnaeus) (Cnidaria, Octocorallia) was studied by incubating colonies with 45calcium (45Ca) or 14C-aspartic acid (14C-asp), and measuring the radioisotope uptake in sectioned fractions including tissue, isolated spicules and axial skeleton. The results show that the ectodermal structure of the coenenchyme was fully able to transport calcium and aspartic acid from base to tip at the same rate. Following short-term incubation, the rate of axial skeleton formation, as measured either with 45Ca or 14C-asp incorporation, was slightly higher at the base of the colony than at the tip. While 14C-aspartic acid deposition in the spicule fraction was maximal at the base of the colony, the distribution of 45Ca in the same fraction showed a U-shape with a maximum at the tip. The total rate of calcification (spicules and axial skeleton) was about 1.6 times higher at the tip of the colonies. A 5-day kinetic experiment showed that the growth process of axial skeleton differed between base and tip. Indeed while the growth rate of axial skeleton was almost constant at the base, a biphasic pattern was observed at the tip suggesting that the apical growth rate of axial skeleton was the consequence of two different calcium supplies: the first one, directly supplied from seawater, and a second one, indirectly supplied via an other pool and significant after 3 days of incubation in labelled seawater. A 1-week pulse chase experiment showed that, while no modification in the distribution of 45Ca occurred among the three compartments (tissue, spicules, axial skeleton) in the basal part of the colony, the skeletal fraction increased at the expense of both spicules and tissue at the apical part. All these results suggest, in accordance with previous studies (Allemand and Grillo [1992] J. Exp. Zool. 262:237-246; Grillo et al. [1993] Mar. Biol. 117:119-128; Allemand et al. [1994] Bull. Inst. Océanogr. Monaco, no. special 14:129-139), that the growth pattern of red coral colony displays important differences between the tip and the rest of the colony. We suggest that the growth of axial skeleton along the colony results from a secretion by the skeletogenic epithelium leading to a centrifugal growth as demonstrated in Corallium johnsoni (Lawniczak, [1987] Senckenberg. Marit. 19:149-161). At the tip of the colony, mechanical incorporation and fusion of spicules into the skeleton are superimposed to this secretion. These results are discussed in the context of Corallium taxonomy

 

[Habib]

One more for keeping you quite a bit longer

I'll be back :wavehand:

Cell Tissue Res 1984;235(3):533-8

Synthesis and transport of the organic matrix of the spicules in the gorgonian Leptogorgia virgulata (Lamarck) (Coelenterata: Gorgonacea). An autoradiographic investigation.

Kingsley RJ, Watabe N.

The sequence of the synthesis and transport of the organic matrix of spicules has been elucidated in the gorgonian Leptogorgia virgulata by use of 3H-aspartic acid as the tracer in electron-microscopic autoradiography. The entire process of matrix synthesis and transport takes approximately 2 h. It seems that the protein moiety of the organic matrix is synthesized in the RER prior to 5 min following the initial 10 min incubation in the tracer. At the 5 min chase the label is moving from the RER to the Golgi complexes where the carbohydrate moiety of the matrix is presumed to be synthesized. At the 5 to 15 min chases the label is transported out of the Golgi complexes via Golgi vesicles. This phase continues for 30 min. From 60 to 120 min the 3H-aspartic acid moves to the spicules. After 120 min the majority of the label has moved into the spicules. Silver grain counts over both multivesicular and electron-dense bodies remain at relatively low and constant levels over 4 h indicating that neither organelle is involved in the synthesis and transport of the organic matrix.

 

[saltshop]

What kinds of things were they saying?

Hehe...well according to some(?) aspartame is responsible for just about every type of cancer there is, Alzheimers Disease, Lou Gehrigs Disease/ALS, confusion and loss of mental ability and fatigue as Habib eluded to, Gulf War Syndrome, and a host of other ailments. Pretty much you name it and aspartame is apparently causing it. Funny thing is, I could swear Lou Gehrig died well before aspartame came out, but maybe he died of some other disease. What did he die from again? They site that aspartic acid, phenylalanine, and menthol the products of aspartame are all neurotoxins which I beleive is probably true at outrageous dosages. They do fail to mention that "healthy" substance such as milk and citrus juice contain ten fold of these aminos. Go to this site and there are many others like it...

http://www.aspartamekills.com

You will see that aspartame was even responsible for O.J. Simpson killing Nicole Brown(toward the bottom of the page)...hey I thought he was innocent???

 

[Habib]

Thanks for the aspartamekills link.
I noticed there Dr. Christine Lydon (halfway the page just above Michael J. Fox). Could she be given a forum over here?

Ok I have added two more abstracts giving I think enough food for tought. One thing which is demonstrated for our purpose is that aspartic acid can be taken up from the surrounding water and be utilized. I have to add that there are papers which have demonstrated that aspartic acid can be synthesized by corals (a.o. from some other amino acids)

Randy,
One of my intention was, in giving data in one of the first posts in this thread, to show rough similarities between corals on one hand and on the other hand the similarity between biotic and abiotic precipitated calcium carbonate in corals (e.g. shells is something different, lots of glycine).

So I'll try summarizing what I've gotten out of this. Thanks for posting it. I don't expect an answer to the questions, but if you have any, or even opinions, I'd be happy to hear them.

So I'll try summarizing what I've gotten out of this. Thanks for posting it. I don't expect an answer to the questions, but if you have any, or even opinions, I'd be happy to hear them. 1. Aspartic acid residues, either as individual amino acids or as parts of proteins, are heavily incorporated into growing CaCO3 crystals from the ocean. It is present for both biological calcification and abiotic calcification in similar amounts. Does this result suggest that it is present in corals "accidently"? That is, that it doesn't imply any special role in corals? Or maybe this incorporation is incidental to the role that organics do play in vivo, but that these are largely not left behind.

presence of aspartic acid in abiotic samples makes it likely that aspartic acid occurs in the water.
And of course has aspartic acid a high affinity for calcium carbonate surfaces, after all it has been attracted to abiotic aspartic acid. Some affinity is easily imaginable based on chemical structure alone.


Since aspartic acid is the major constituent of the organic matrix in corals, and it is recognized widely that organic matrices associated with bio-minerals are higly essential for bio-mineralization, it is very likely that aspartic acid is not there just by accident but is highly essential (see also last two abstracts).

2. That the role for organics in biological calcification could be to:

a. Prevent crystal poisoning by Mg++, phosphate, and other organics.
b. Direct crystal growth to regions where it is most desired.
c. To hold loose assemblies of crystals together in the skeleton.
d. Something else?


a. I don't think prevent crystal poisoning but decrease the effect of PO4 and similar acting substances. Depending on conectartion of course.

b + c . That has been postulated by some authors and even that it forces the araogonite crystallomorph. Others say it terminates crystallisation.
I think that it envelopes the macro-crystals, the outer surface of the organic matric facilitating further growth of macro-crystallites.

d. Yes. To keep us busy?

3. That since aspartic acid naturally binds strongly to CaCO3 crystals, that is an obvious "choice" for organisms to use in attaching organics to CaCO3 surfaces.

Could be, might perhaps be also alternatives. Things like availability, transportability and other factors also play an important role. But I think nature is far more clever than we are.

4. That aspartic acid may be present in the ocean and aquaria because it appears to be attached to abiotic CaCO3, but that it may still not be present in high enough concentrations for optimum growth rates. Maybe it's not present in high enough concentrations precisely because it is bonded to CaCO3 and not floating around the aquaria as much as other amino acids?

I think very likely as long we don't know what the exact source of aspartic acid is.

Would aspartic acid be beneficial as an additive?
For me yes because I earn money with it.

I personally found the information on abiotic calciumcarbonate very important some 8 or so years ago. It meant that aspartic acid occured in the water. Also it would make further growth of calcium carbonate easier if nasty crystal poisons would be present.

Also if we look at the importance of organic matrices, aspartic acid being a major constituent in it , proof that it can be absorbed from the water, is brought to the point where it should be, any excess broken down by bacteria,....

So: Yes very useful as an additive unless the aspartic acid concentration + the concentration of any of it's precursor is high enogh then probably no need to add aspartic acid.

One product of which I know contains aspartic acid (but also some other things) is used by a.o. very critical coral farmers. Reactions are faster growth and intense coloration (both not scientifically documented!!!). BUT tanks which have some other problems don't show these positive effects. So aspartic acid is defenitely is not a miracle. But I think it is limiting in some/many tanks. The more parameters are understood and corrected the more parameters will be discovered that are limiting then.

 

[Randy Holmes-Farley]

For me yes because I earn money with it.

:lolspin:

OK, now that I've stopped laughing, what product of yours has the aspartic acid? Unfortuantely, I'm not intimately familiar with your entire product line.

 

[garbled]

Both of the abstracts posted by habib seem to discuss octocorals, and the use of aspartic acid in spicule growth.

That makes me wonder if there is a specific reason they chose octocorals for these studies, over, say an acropora.

It would be interesting to try growth tests with this stuff, unfortunately I don't have the space (yet) to do so. Perhaps two aquariums, side by side, equal in all respects but additions of the acid to one.

This has been a very fun subject!

 

[andy-hipkiss]

And just to put you on the spot Habib, would you care to differentiate between Salifert AminoAcids (is Bio Coral a current product? if so what's the difference?) and Marine De Luxe?

Maybe this is beyond the scope of this forum (?) so an email would be most appreciated if Randy is uncomfortable with too many product related Q's.

 

[Randy Holmes-Farley]

Maybe this is beyond the scope of this forum (?) so an email would be most appreciated if Randy is uncomfortable with too many product related Q's.

No, discussion of products is always welcome, especially if we might get information that is not generally available

The only limitations would kick in if people started posts just to hawk a product.

 

[EricHugo]

Hi Randy and others:

I was sent a link to this thread as being particularly interesting, and I agree. It is extremely interesting. I was asked to add anything I might be able to, but the people involved here have way more information on this subject than I do. I was very impressed with this thread. One thing I would mention, that really doesn't change anything that much, is that much of the "organic matrix" in coral skeletons and other aragonite may not be direct precipitation but be composed of other organisms. In many coral skeleton studies, there may be a methods issue. Coral skeleton is usually stripped of tissue and then ground up for analysis. Within this material are endolithic algae, fungi and bacteria, all of which may tend to confound results as this organic matrix may be confused with anything directly incorporated by the polyp.

 

[saltshop]

what product of yours has the aspartic acid? Unfortuantely, I'm not intimately familiar with your entire product line.

Andy mentioned them above- "Bio Coral" and "Marine De Luxe"...Thanks! I see Marine Depot carries the Bio Coral. I will be placing an order from them in the next week or so anyway...figure will give it a whirl, why not? Will only add it to the 10+ yr. tank as I am very familiar with that one and any change can be noticed "fairly" easily as nothing in the routine for the tank has changed in several years other than changing the bulbs once a year.

Is it even possible to test water samples for amino acids such as this? Just curious....

 

[Randy Holmes-Farley]

It would be straightforward to test for aspartic acid with a modern chromatography instrument of some type (HPLC, ion chromatography, etc). Without such a device, I suspect that trying to select one of the amino acids is less than trivial in the presence of others.

 

[Habib]

Andy mentioned them above- "Bio Coral" and "Marine De Luxe"...

Marine deluxe is not ours and I don't know if it contains aspartic acid.

No, discussion of products is always welcome, especially if we might get information that is not generally available

Thanks, Randy.
I like to be on this Forum as a hobbyist and not as a Manufacturer except in those cases in which I might be of some help to fellow hobbyists. Even then I try to be as objective as possible (I hope).



And just to put you on the spot Habib, would you care to differentiate between Salifert AminoAcids (is Bio Coral a current product? if so what's the difference?) and Marine De Luxe?

Since more people have asked and it does not seem to be a problem if I comment on this....

It started as Organo-Plus then it was called Bio-Coral and now it is called Coralline AminoAcids (Amino Coral in non english speaking countries).

Besides a change in name some improvements have been made. Especially Bio-Coral was prone to going bad occasionally(growth of fungi) so now and then we had to take it back and throw it away.

Coralline AminoAcids is better preserved with naturally occurring substances. Approx. 40% of the active ingredients is aspartic acid.

We keep the formulation secret so if you really want to know what you are adding then this product is of no use. There definitely is a lot of water in it but a few components (not aspartic acid) are at 50% solubility limit (there even will be now and then some particles visible).

I am quite willing to ship a limited number of concentrates (500 ml to be diluted to 1000 ml) to those who seriously want to experiment with it. :bounce1:

 

[Habib]

Garbled,

Both of the abstracts posted by habib seem to discuss octocorals, and the use of aspartic acid in spicule growth.

That makes me wonder if there is a specific reason they chose octocorals for these studies, over, say an acropora.


Organic matrix synthesis in the scleractinian coral stylophora pistillata: role in biomineralization and potential target of the organotin tributyltin
D Allemand, E TambuttE, JP Girard and J Jaubert
Observatoire Oceanologique Europeen, Centre Scientifique de Monaco, Avenue Saint Martin, MC-98000 Monaco, Principality of Monaco, Commissariat a l'Energie Atomique - LDG, BP 12, F-91680 Bruyeres-Le-Chatel Cedex, France and Laborat.

The kinetics of organic matrix biosynthesis and incorporation into scleractinian coral skeleton was studied using microcolonies of Stylophora pistillata. [14C]Aspartic acid was used to label the organic matrix since this acidic amino acid can represent up to 50 mol % of organic matrix proteins. External aspartate was rapidly incorporated into tissue protein without any detectable lag phase, suggesting either a small intracellular pool of aspartic acid or a pool with a fast turn-over rate. The incorporation of 14C-labelled macromolecules into the skeleton was linear over time, after an initial delay of 20 min. Rates of calcification, measured by the incorporation of 45Ca into the skeleton, and of organic matrix biosynthesis and incorporation into the skeleton were constant. Inhibition of calcification by the Ca2+ channel inhibitor verapamil reduced the incorporation of organic matrix proteins into the skeleton. Similarly, organic matrix incorporation into the skeleton, but not protein synthesis for incorporation into the tissue compartment, was dependent on the state of polymerization of both actin and tubulin, as shown by the sensitivity of this process to cytochalasin B and colchicin. These drugs may inhibit exocytosis of organic matrix proteins into the subcalicoblastic space. Finally, inhibition of protein synthesis by emetin or cycloheximide and inhibition of N-glycosylation by tunicamycin reduced both the incorporation of macromolecules into the skeleton and the rate of calcification. This suggests that organic matrix biosynthesis and its migration towards the site of calcification may be a prerequisite step in the calcification process. On the basis of these results, we investigated the effects of tributyltin (TBT), a component of antifouling painting known to interfere with biomineralization processes. Our results have shown that this xenobiotic significantly inhibits protein synthesis and the subsequent incorporation of protein into coral skeleton. This effect was correlated with a reduction in the rate of calcification. Protein synthesis was shown to be the parameter most sensitive to TBT (IC50=0.2 micromol l-1), followed by aspartic acid uptake by coral tissue (IC50=0.6 micromol l-1), skeletogenesis (IC50=3 micromol l-1) and Ca2+ uptake by coral tissue (IC50=20 micromol l-1). These results suggest that the mode of action of TBT on calcification may be the inhibition of organic matrix biosynthesis