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Offlinekoopa_troopa
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Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor
    #24649145 - 09/21/17 02:09 PM (6 years, 6 months ago)

Greetings and salutations fellow fungi farmers! I've been absent from the community for quite some time due due to my endeavours with truffle cultivation.
   
    During the past year I have made significant discoveries and breakthroughs developing specialized compost, and was able to create a compost the nearly doubled the myceliums colonization rate on the hazelnut sapling roots that is the highest percentage ever recorded that remained for an extended period.
 
    Now that I've reached a  point where I have some free time,  I've taken my knowledge and experts I developed while customizing compost for truffles and applied it to saprophobic fungi.
 
    During this time I have made great strides and discoveries creating high end premium customized compost with many additives that have been scholarly peer reviewed proven with very high statistical confidence levels and consistent yields over 20% higher than the control group. And the same goes for all seven additives which when combined with a customized compost specifically tailored to promote mycelium colonization and full consistent flushes.
 
    I am very interested in becoming a sponsor here at the shroomery. And I am willing to give out 20 free samples to experienced growers to prove it's efficacy. But first and foremost I am posting this to gauge the communities interest in such a product. Now this is a super premium product, comparable  to high end nutrients for hydroponic marijuana grows. The free samples I give out will prove without a doubt it's ability to exponentially increase yields.. and for all the doubters out there, I would be more than happy to provide complete transparency and post all the scientific articles for each ingredient. The price of this compost will be high (but not unreasonable and it's quality will more than justify the cost).  Keep in mind not only am I having to create massive amounts of custom tailored compost but also having to deal with the high cost of purchasing very large amounts of all these materials in bulk.
 
  I know for a fact from experience  and literally hundreds  of grows with this product, that once everyone sees the proof that using this compost will exponentially increase yields, and I will even guarantee these results or your money back, no questions asked! o
   
  Please feel free to ask any questions and please let me know if you would be interested in such a product, so I know if I should go forward or not. Thanks guys!


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Koopa Out
OfThisWorld,SmilingDownOnOurs: BirsdEyeView,NeverNewLife,PastsFastAsCars

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InvisibleZiran
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: koopa_troopa]
    #24649152 - 09/21/17 02:10 PM (6 years, 6 months ago)

pictures?


--------------------
Song Of Healing
:super: Updated Pf Tek Guide :super:
Ziran's Teks
AMU Q&A Thread
The Chinese word for nature is zìrán and it means that of which is of itself.


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Offlinekoopa_troopa
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Ziran]
    #24649255 - 09/21/17 02:41 PM (6 years, 6 months ago)

I've been using a very similar variation for quite some time, if you check my images on my profile you can see some penis envies I grew with a similar compost. Unfortunately when I was developing the most recent and most advanced version, I had no plans on selling it and was doing it mainly to satisfy my scientific curiosity. But I have some colonized spawn that I'm going to do a comparison grow with and should be ready within a month. But once you see my images and what was capable with a much more inferior version, you will immediately get a good idea of it's potential as that grow was done simply from multi spore inoculation yet still produced world class specimens.


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Koopa Out
OfThisWorld,SmilingDownOnOurs: BirsdEyeView,NeverNewLife,PastsFastAsCars

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InvisiblePastywhyteMDiscord
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: koopa_troopa]
    #24649312 - 09/21/17 03:01 PM (6 years, 6 months ago)

What is the BE on average then? I must be blunt, while you have some above average sized fruits, your pinset is poor which in itself could explain the larger fruit body.

Nothing I see there makes me want to use your substrate. More detail would be nice.

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InvisiblemushboyMDiscord
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Pastywhyte] * 1
    #24649354 - 09/21/17 03:18 PM (6 years, 6 months ago)

Those pe did look like dildos...  But i refuse to believe you made super compost worth buying when you cant even keep your room clean. 

My loss i guess. 

But i would like more details as to whats in your special sauce:begger:

Its all about that sauce.

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OfflineVroomerMcZoomers
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: koopa_troopa]
    #24649365 - 09/21/17 03:25 PM (6 years, 6 months ago)

I just wrote a post this morning asking about where I could get good compost...

Im interested, who wouldn't be interested here?


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Either you repeat the same conventional doctrines everybody is saying, or else you say something true, and it will sound like it's from Neptune.  -Noam Chomsky

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InvisiblemushboyMDiscord
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: VroomerMcZoomers]
    #24649367 - 09/21/17 03:26 PM (6 years, 6 months ago)

For the same reason i stated in your post:shrug:

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OfflineVroomerMcZoomers
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: mushboy]
    #24649381 - 09/21/17 03:29 PM (6 years, 6 months ago)

What?  You tried compost and weren't all that impressed, this guy says his compost works like gangbusters.

What am I to think? 

Im going to have to try it myself or Ill never know, I guess.


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Either you repeat the same conventional doctrines everybody is saying, or else you say something true, and it will sound like it's from Neptune.  -Noam Chomsky

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InvisiblemushboyMDiscord
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: VroomerMcZoomers]
    #24649388 - 09/21/17 03:32 PM (6 years, 6 months ago)

Nooo... This..

Quote:

a brick of compressed coocunt husks will outperform or at least be on par with all of those



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OfflineVroomerMcZoomers
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: mushboy]
    #24649397 - 09/21/17 03:36 PM (6 years, 6 months ago)

Oh, yeah.

I have never used coir.  But Im going to try it next time for sure.  Do you have a favourite tek?  Im kinda confused about it.


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Either you repeat the same conventional doctrines everybody is saying, or else you say something true, and it will sound like it's from Neptune.  -Noam Chomsky

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InvisibleJosex
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: koopa_troopa]
    #24649406 - 09/21/17 03:40 PM (6 years, 6 months ago)

Quote:

The price of this compost will be high (but not unreasonable and it's quality will more than justify the cost).



Even if it's true your compost can increase yields by 20% it would still be way cheaper to just do more tubs using coir, simple as that :shrug:

I'm very skeptical anyway, mushrooms are 90% water, so the main purpose of a bulk substrate is to suply that water. How's your compost going to help with that? Cubensis mycelia is already very good at transporting the water available for the formation of fruit bodies, is your compost going to  magically improve the mycelium's ability to use the water available? Or improve pinsets? Make the mushrooms bigger? The only thing that can really affect those is genetics, and as far as we know ain't no friking additive out there that can do that.

Edited by Josex (09/21/17 03:50 PM)

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InvisiblePastywhyteMDiscord
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Josex]
    #24649782 - 09/21/17 05:37 PM (6 years, 6 months ago)

MS, coir verm, 5 quarts spawn. Cost of materials for this tub was like 6 bucks including the grain and trash bag. Why do I need to pay more than that?


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OfflineVroomerMcZoomers
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Pastywhyte]
    #24649793 - 09/21/17 05:42 PM (6 years, 6 months ago)

What's MS?


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Either you repeat the same conventional doctrines everybody is saying, or else you say something true, and it will sound like it's from Neptune.  -Noam Chomsky

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OfflineCodeinecowboy
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: VroomerMcZoomers]
    #24649805 - 09/21/17 05:47 PM (6 years, 6 months ago)

Multispore

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Offlineinvitro

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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: koopa_troopa]
    #24649820 - 09/21/17 05:52 PM (6 years, 6 months ago)

Quote:

koopa_troopa said: and for all the doubters out there, I would be more than happy to provide complete transparency and post all the scientific articles for each ingredient.




Ok then, please do post the articles!

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InvisibleBoogieman47
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Codeinecowboy]
    #24649824 - 09/21/17 05:54 PM (6 years, 6 months ago)

The genetics of your pe are pretty killer though probably some of the better looking ones ive seen


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InvisiblebodhisattaMDiscordReddit
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Boogieman47]
    #24649873 - 09/21/17 06:08 PM (6 years, 6 months ago)

Quote:

. and for all the doubters out there, I would be more than happy to provide complete transparency and post all the scientific articles for each ingredient




you would have just put it in the post then

Quote:

VroomerMcZoomers said:
Im interested, who wouldn't be interested here?




anyone who's been around the block

Quote:

VroomerMcZoomers said:
Oh, yeah.

I have never used coir.  But Im going to try it next time for sure.  Do you have a favourite tek?  Im kinda confused about it.




the tek is on the brick, it says mix with water in a bucket.
we just use hot(boiling) water instead.

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Offlineverum subsequentis
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Boogieman47]
    #24649884 - 09/21/17 06:13 PM (6 years, 6 months ago)

I'd love to try it. I'm always skeptical about unproven shit but if it works I'll help you prove it. I'll do two tubs of the same iso, one with your shit and one cvg, if you send me some. I don't have anything to lose.

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Offlineverum subsequentis
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Pastywhyte]
    #24649892 - 09/21/17 06:15 PM (6 years, 6 months ago)

Quote:

Pastywhyte said:
MS, coir verm, 5 quarts spawn. Cost of materials for this tub was like 6 bucks including the grain and trash bag. Why do I need to pay more than that?






Beautiful knife pasty.

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Offlinekoopa_troopa
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Re: Developed a super compost exponentially increases yields, want to gauge interest before bc a vendor [Re: Pastywhyte] * 2
    #25022117 - 02/25/18 10:44 PM (6 years, 1 month ago)

You know I nearly never even replied to this thread due to the complete and utter hate spewed ignorance of those that have had some decent Cube grows... lol its hilarious hearing some of this. Hating on the uneven pinset of a MUTANT genetic freak of a strain?!? There is no such thing as a completely even PE pinset that remains that way until maturation, as the largest strongest fruitbodies outcompete them for resources.

The comment about mushrooms not benefiting from the addition of organic compounds and nutrients because they are 90%, so are you so why dont you follow that logic and see how well you do off a purely vitamin and sugar diet :smile:

The comment about my dirty room, check the date bro, that was over 12 years ago when I was a sophomore and college... 

I am a professional mycology consultant, pioneering breakthrough truffle production research utilizing data science, machine learning, deep convolutional and deep neural networking AI in conjuntion with rover drones and Smart field implants.

If you believe your cubes can be all they can be by growing on any amendmant with water retention then compare ones grown solely on verm to high end compost ameliorated with agricultural waste such as cotton gin waste, rape seed oil, activated charcoal, urea  etc,.. now this is may seem slight overkill for simple cubes, but it will give you exceptional yields with potency like nothing you have encountered.

The specialty compost I was suggesting, is ideally for gourmet and medicinal mushrooms. If you are interested in fungi nutrient metabolism. Here is a breakdown of primary fungal metabolism:

Metabolic pathways which characteristically operate when a fungus is growing at or near its maximum rate are described as primary pathways; secondary pathways become operational (or amplified) when growth rate is limited in some way to a level below the maximum (Bu’Lock, 1967). The fundamental function of primary metabolism is the utilisation of nutrients to form ATP and reduced nucleotide coenzymes (NADH+ and NADPH+), which together equal chemical energy and reducing power, and the compounds which serve as precursors of cellular components, especially macromolecular constituents. This section will be concerned with aspects of metabolism responsible for formation of substrates which serve biosynthetic pathways and compounds providing energy and reducing power; the so-called primary or intermediary metabolism.
The major source of energy and reducing power is the catabolism of carbohydrate. Although other carbon-containing compounds can be utilised for these purposes by most cells, the full sequence of enzymic processes are conventionally represented as involving the controlled release of energy by the use of atmospheric oxygen to convert glucose to CO2 and water. This overall process is described as respiration and its chemically balanced (or stoichiometric) summary equation is:
C6H12O6 + 6O2 → 6CO2 + 6H2O + energy
Note that this equation does not even begin to describe the biochemical mechanisms which achieve the indicated chemical transformation, but it does emphasise that for the conversion of each mole of glucose, six moles of oxygen must be absorbed from the atmosphere, and six moles of CO2 and six moles of water appear within the cell, and 2 900 kJ of free energy (i.e. energy capable of doing some work) are released. To put this into more readily grasped units, respiration of 1 g of glucose uses 1.07 g (747 cm3) oxygen and produces 1.47 g (747 cm3) CO2 and 0.6 g water, releasing 16.1 kJ of energy.
To achieve this basic chemistry the living cell uses a sequence of enzymically controlled reactions. These are conveniently divided into three phases or subpathways:
glycolysis including the pentose phosphate pathway (PPP),
the tricarboxylic acid (TCA, or Krebs) cycle,
oxidative phosphorylation.

The word glycolysis describes the conversion of glucose to pyruvate without implying a particular pathway. In fact, there are three enzymic pathways which might be used, though one does tend to predominate. The Embden-Meyerhof-Parnass (EMP) pathway is the major pathway in most species; it comprises nine enzymic steps, all of which occur in the cytoplasm (Fig. 7). The net outcome of the reactions summarised in Fig. 7 is that one molecule of glucose is converted to two molecules of pyruvic acid plus two molecules of ATP and two molecules of NADH2. Thus, the energy yield (2ATP + 2NADH2, since the latter do represent potential chemical work), is rather small; the main function of the EMP pathway being conversion of glucose to pyruvate for processing in the TCA cycle.

A commonly encountered alternative glycolytic pathway is the pentose phosphate pathway (PPP) (Fig. 8). This is also called the hexose monophosphate pathway (HMP) and in strictly chemical terms such a name is accurate; glucose 6-phosphate is diverted out, undergoes a range of chemical conversions, and then fructose 6-phosphate and glyceraldehyde 3-phosphate feed back into the EMP pathway. But the name pentose phosphate pathway does acknowledge that the PPP provides pentose sugars for nucleotide synthesis (which includes coenzymes and energy carriers as well as RNA and DNA), erythrose phosphate for the synthesis of aromatic amino acids through the shikimic acid pathway, and NADPH2, which is the coenzyme most often used in biosynthetic reactions that require reducing power, especially fat and oil synthesis. So although the PPP can theoretically achieve complete glycolysis (six cycles through the reaction sequence would completely oxidise a molecule of glucose to CO2), it is more likely to be involved in furnishing biosynthetic intermediates. The PPP also, of course, provides a route for utilisation of pentose sugars which become available as carbon sources, and for interconverting hexose and pentose phosphates.

ere is a third glycolytic pathway, the Entner-Doudoroff (ED) pathway (not illustrated here) that proceeds via 6-phosphogluconate to 2-keto-3-deoxy-6-phosphogluconate, which gives rise to pyruvate and glyceraldehyde 3-phosphate. It is a common glycolytic pathway in bacteria, but has been demonstrated in only a few fungi.
The use of different glycolytic pathways in any cell will reflect the relative contribution their intermediates are required to make to the functions of the cell; they will change with age, activity and nutrition. In general, since the PPP provides intermediates for biosynthesis, use of this pathway increases in rapidly growing and in differentiating cells, and is minimised in those which are resting or quiescent.

Whichever glycolytic pathway contributes the pyruvate, this latter molecule is formed in the cytoplasm and must then be transported into the mitochondrion where it is converted to acetyl coenzyme A (acetyl-CoA). This step is achieved by the pyruvate dehydrogenase complex; a combination of enzymes which first decarboxylate pyruvate and then transfer the resulting acetyl group to coenzyme A.
The TCA cycle is cyclic because its ‘end-product’, oxaloacetate, reacts with acetyl-CoA to introduce what remain of the pyruvate carbon atoms into a reaction sequence (Fig. 9) the primary function of which is to convert pyruvate formed in glycolysis entirely to CO2, the released energy being captured primarily in NADH2. The overall stoichiometry is that one molecule of pyruvate with three molecules of water forms three molecules of CO2 and releases 10 protons, the latter appearing in the form of three molecules of NADH2, and one each of FADH2 and the ‘high energy’ compound GTP. The succinate dehydrogenase enzyme is bound to the inner mitochondrial membrane (and, because of this, is often used as a marker for the presence of mitochondria in fractionated cell extracts), the other enzymes of the TCA cycle occur in the mitochondrial matrix.

A common variant of the TCA cycle is the glutamate decarboxylation loop in which 2-oxoglutarate is aminated to glutamate rather than being oxidatively decarboxylated to succinate. The glutamate is decarboxylated to 4-aminobutyrate; transamination between the latter and 2-oxoglutarate yielding succinate semialdehyde which, on oxidation, feeds back into the TCA cycle as succinate (central panel in Fig. 9). The enzymes of this loop have been found to be at high activity in fruit bodies, especially caps, of the ink cap mushroom Coprinopsis cinerea, where it is the normal route of TCA metabolism in this organism. The glutamate decarboxylation loop also operates in Agaricus bisporus.
Through glycolysis and the TCA cycle, all of the carbon contained in the substrate glucose is released as CO2. However, very little energy is released, most of it being captured in NADH2 (with a small amount in GTP and FADH2). The energy represented in the form of the reduced coenzymes is recovered as ATP through the electron transport chain, located on the inner mitochondrial membrane, in the process known as oxidative phosphorylation. The electron transport chain transfers electrons from the reduced coenzymes through a series of reactions until the electrons are finally passed to oxygen, reducing it to water. Stepwise transfer of electrons between components of the electron transport chain leads to the pumping of protons from the mitochondrial matrix into the intermembrane space.
The resulting proton gradient (the pH in the intermembrane space is about 1.4 pH units lower than that of the matrix) is used to generate ATP. Transfer of a pair of electrons from one molecule of NADH2 to oxygen leads to proton pumping at three sites in the chain, at each of which the consequent proton gradient can be used to synthesise one molecule of ATP. The ATP is synthesised by an enzyme complex located on the matrix side of the inner mitochondrial membrane. As protons move down a channel in this complex (the channel, known as the F0 sector, is composed of at least four hydrophobic subunits forming the proton channel located in the membrane) the associated F1 sector (containing five different subunits) projects into the matrix and is responsible for ATP synthesis.

Although we are primarily concerned here with catabolism, the pathways described permit sugar synthesis with just a few modifications. We stressed above that glycolysis and the TCA cycle provide opportunities for the fungus to make use of a very wide range of potential carbon and energy sources; but one which is successfully growing on acetate, for example, is clearly required to synthesise all of those compounds which have more than two carbon atoms chained together. In such circumstances glycolysis cannot simply be reversed because the steps governed by kinases (hexokinase, phosphofructokinase and pyruvate kinase) are irreversible, so for these steps in particular, additional enzymes are required for gluconeogenesis
.
In these circumstances the first steps in conversion of pyruvate to carbohydrate are carried out by pyruvate carboxylase, which synthesises oxaloacetate which is then decarboxylated and phosphorylated to phosphoenolpyruvate by phosphoenolpyruvate carboxykinase. The phosphoenolpyruvate can then be converted to fructose 1,6-bisphosphate by reversal of the EMP pathway, but an additional enzyme, fructose bisphosphatase, is required to generate fructose 6-phosphate. As the sugar phosphates are readily interconvertible, once this compound is formed oligosaccharide and polysaccharide synthesis can proceed. The structures of many of the polysaccharides formed have been shown earlier in this chapter.

Glycolysis and gluconeogenesis are obviously alternatives which demand close control to assure metabolic balance. Phosphofructokinase is the key glycolytic control point, and fructose bisphosphatase responds inversely to the same molecules, being ing allosterically activated by citrate but inhibited by AMP.

Before leaving carbohydrate metabolism, it is worth mentioning here that the sugar alcohol mannitol and the disaccharide trehalose are almost always found among the water-soluble cytoplasmic carbohydrates in fungi; trehalose being the most widely-distributed sugar in fungi. Mannitol and trehalose seem to serve as transient storage compounds (i.e. molecules capable of immediate mobilisation when required) and both have been identified as substrates used for the metabolism associated with spore germination.
Trehalose synthesis/accumulation/degradation cycles occur at a number of stages in development of a fungus so it is quite clear that this sugar is the ‘common currency’ of the fungal carbohydrate economy. It is synthesised, as trehalose 6-phosphate, by the enzyme trehalose phosphate synthase from glucose 6-phosphate and the sugar nucleotide UDP-glucose. There may be other functions for mannitol. It can certainly serve an osmoregulatory function in the marine fungus Dendryphiella and may serve the same purpose in fruit bodies of the cultivated mushroom, Agaricus bisporus, in which it can be accumulated to concentrations of up to 50% of the total dry weight (yes, that’s right, half of the cultivated mushroom on your plate is mannitol); similar concentrations have been encountered in fruit bodies of Lentinula edodes (shiitake). In A. bisporus, mannitol is synthesised by reduction of fructose by an NADP-linked mannitol dehydrogenase.
Fats are molecules of glycerol in which the three hydroxyl groups are replaced with three fatty acid molecules. In degradation the first step is carried out by lipase which removes the fatty acids from the glycerol. The latter can be converted to glyceraldehyde 3-phosphate and thereby enter glycolysis (see Fig. 8), but it represents only about 10% by weight of a fat molecule, the bulk being represented by the fatty acids which consist of long carbon chains (e.g. palmitic acid, C16; stearic acid, C18).
These chains are degraded by sequential removal of the two terminal carbon atoms in the form of an acetyl group attached to CoA (Fig. 11). Because the cleavage occurs at the second (β) carbon atom, this process is called β-oxidation and it takes place in the mitochondrial matrix. Each such cleavage is oxidative, enzymes passing the H-atoms to the coenzymes NAD and FAD. Thus, degradation of palmitic acid requires seven cleavages and yields eight molecules of acetyl-CoA (which enter the TCA cycle), seven NADH2 and seven FADH2 (both of which enter the electron transport chain for oxidative phosphorylation). Oxidation of fatty acids releases considerable amounts of energy; for example, one molecule of palmitic acid will give rise to about 100 molecules of ATP. This is why fats are such effective energy storage compounds.
Ultimately, all nitrogen in living organisms is derived from the native element in the atmosphere. Each year an amount between 100 and 200 million tonnes of atmospheric nitrogen is reduced to ammonium by the nitrogenase enzyme system of nitrogen-fixing bacteria and blue-green algae. On the basis of present knowledge, it is unlikely that any fungi are able to fix elemental nitrogen.

a fungus is unable to access amino groups by direct absorption of amino acids, they have to be formed and the most immediate source is by the assimilation of ammonium. The only route of ammonium assimilation which can be considered pretty well universal in fungi is the synthesis of glutamate from ammonium and 2-oxoglutarate by the enzyme glutamate dehydrogenase. Many filamentous fungi and yeasts have been shown to produce two glutamate dehydrogenase enzymes; one linked to the coenzyme NAD and the other linked to NADP.
As can be appreciated from Fig.12, the interconversion of 2-oxoglutarate and glutamate is a reaction which occupies a central position in metabolism and is one at which important pathways in both carbon metabolism and nitrogen metabolism come together. The reaction is readily reversible and it is often considered that NAD-linked glutamate dehydrogenase (NAD-GDH) has a deaminating or catabolic role (glutamate → 2-oxoglutarate + ammonium), while the NADP-linked enzyme provides the aminating or anabolic function (2-oxoglutarate + ammonium → glutamate). However, some organisms have evolved different patterns of endogenous regulation, especially in relation to their morphogenetic processes. For example, in the ink cap mushroom Coprinopsis cinerea the NADP-GDH normally appears at high activity only in the cap tissue of the mushroom fruit body where it is located in the basidia apparently protecting meiosis and sporulation from inhibition by ammonium, i.e. acting as an ammonium detoxifier rather than ammonium assimilator.
This example aside, NADP-GDH is generally the most important enzyme involved in ammonium assimilation in mycelia and its activity is often increased when ammonium is provided as a growth-limiting, sole nitrogen source. However, in some fungi an alternative enzyme system appears to scavenge for ammonium when it becomes limiting, this is the glutamine synthetase/glutamate synthase system. Glutamine synthetase is widely, perhaps universally, distributed and is responsible for synthesis of glutamine. However, glutamine synthetase can have a high affinity for ammonium and, in combination with glutamate synthase (which converts glutamine + 2-oxoglutarate to two molecules of glutamate), forms an ammonium assimilation system which can recover ammonium even when this molecule is present at extremely low concentrations. The net result (2-oxoglutarate + NH4+ → glutamate) is the same as the reaction promoted by NADP-GDH, but the cost is higher as glutamine synthetase uses ATP to make glutamine. The glutamate synthase mechanism is common in bacteria and is encountered in fewer fungi, but has been demonstrated in Neurospora crassa, Aspergillus nidulans and several yeasts and mycorrhizal fungi.
Some yeasts and a larger number of filamentous fungi can utilise nitrate as sole source of nitrogen. Chemically, nitrate is first converted to nitrite which is then converted to ammonium, but the enzymic steps are quite complicated. The complexity of the reaction is reflected in the large number of mutant genes, in both Neurospora and Aspergillus, which have been found to affect nitrate assimilation. The first stage is performed by nitrate reductase which, generally in fungi, has a cofactor containing molybdenum and requires NADPH (NADH in at least some yeasts). Nitrate is thought to bind to the molybdenem-cofactor of the enzyme prior to being reduced by removal of an oxygen atom. Removal of this allows the nitrate formed to be bound to nitrite reductase, through its nitrogen atom, for the reduction to ammonium. The ammonium formed from nitrate is immediately used for the reductive amination of 2-oxoglutarate to glutamate.
Conversion of NO3- to NH3 is a chemical reduction requiring considerable energy expenditure. In fact the equivalent of four NADPH2 molecules (880 kJ of energy) are used to reduce one NO3- ion to NH3 , which is additional to the energy demand for assimilation of the ammonium (one NADPH2 is used for assimilation via NADP-GDH, 1 NADPH2 + 1 ATP for assimilation through glutamine synthetase and glutamate synthase). Given these additional energy demands, it is not surprising that the nitrate reduction machinery is produced only when nitrate is the sole available source of nitrogen, being induced by nitrate and rapidly repressed by the presence in the medium of ammonium or any alternative sources of reduced nitrogen.
The constituents of living cells are in a continual state of flux; all components being subjected to turnover as old materials are catabolised and new ones synthesised. When proteins and other nitrogen-containing compounds are broken down, either as part of this turnover process or as externally supplied nutrients, the carbon can be disposed of as CO2, hydrogen as water and nitrogen either as ammonium or as urea. The use of protein as a carbon source has been discussed above. In these circumstances the organism (animal, plant or fungus) suffers an excess of nitrogen and must excrete it. Experiments with the basidiomycetes Agaricus bisporus, Coprinopsis cinerea and Volvariella volvacea have shown that one third to one half of the nitrogen contained in the protein given as substrate is excreted as ammonium into the medium.
In terrestrial mammals metabolising protein the toxicity of ammonium is avoided by excretion of urea formed through the urea cycle (Fig. 13). However, the enzyme urease seems generally to be constitutive in fungal mycelia so any urea formed is likely to be dissimilated to NH3 and CO2. Nevertheless there are circumstances in which fungi accumulate urea (at which time they repress the urease). Especially large accumulations have been found in fruit bodies of Basidiomycota, where it seems likely that it acts as an ‘osmotic metabolite’ controlling water entry into cells during expansion of the fruit body tissues. Thus, the capacity to synthesise, and even accumulate, urea is well developed in fungi but it seems that it is ammonia which is excreted to dispose of excess nitrogen.

ALSO: google research on SACing (supplementation after colonization!)

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