Nootropics

Lion’s Mane Mushroom: The Journey To Connect Modern Science With Traditional Chinese Medicine

Lion’s Mane Mushroom: The Journey To Connect Modern Science With Traditional Chinese Medicine

The Problem With Mycelium

Lion’s Mane fruiting body powders and extracts were already firmly on our menu. Around this time, the nootropics subreddit, r/nootropics was thriving and it was also rive with research studies and discussions surrounding erinacine A. At the time, it was somewhat unclear if lion’s mane fruiting bodies would contain erinacine A, and slowly but surely it became clear that the fruiting bodies did not really contain a large amount of erinacine A. As a response to these discussions, lion’s mane mycelium products started to pop up from multiple different vendors, some even claiming a total erinacines content of over 4%! Naturally, our customer base started to inquire why we did not carry a lion’s mane mycelium product. Afterall, erinacine A was the talk of the town,with excited redditors and opportunistic marketing specialists doing a fantastic job of making it appear that you could only gain cognitive benefits from erinacine A containing lion’s mane mycelium. Naturally, we immediately started to investigate whether an erinacine A containing lion’s mane mycelium product would be possible. Unfortunately, we immediately hit a roadblock, an erinacine A reference standard did not exist! For those unaware of how analytical testing works, you can’t just put a piece of lion’s mane mycelium in, and have the analytical instrument tell you exactly how much erinacine A is in it. First, you need a sample of pure erinacine A, this is then fed into the analytical instrument. For erinacine A, this ended up being our Waters UPLC. Once the reference standard has been analyzed, you end up with a chromatogram which shows a clear single peak.

Lion’s Mane Mushroom: The Journey To Connect Modern Science With Traditional Chinese Medicine

Next, you run a sample of lion’s mane mycelium through the UPLC, and on the chromatogram you will see a bunch of peaks, one of which will line up with the peak of pure erinacine A, if the lion’s mane mycelium contains erinacine A.

This means that without a reference standard, there really is no way of analyzing the erinacine A content of lion’s mane mycelium. So you can imagine we were really scratching our heads trying to figure out how some of our competing vendors were claiming to have 4% erinacines in their lion’s mane mycelium product. At the time we were still somewhat unaware of how rampant testing fraud is in the supplement industry, so we didn’t immediately put two and two together.

It took almost a year of research, sourcing, and analysis attempts to arrive at the conclusion that fake dry labs were in rampant use throughout the industry.

The way we got real world confirmation of this, was by sending a suspected dry lab a sample of completely spoofed “lion’s mane mycelium”. We used our functional yeast extract to fake the beta-glucans normally present in lion’s mane mycelium, and then we spiked our functional yeast extract with abietic acid. Abietic acid, just like erinacine A, is a diterpenoid. We now had a sample of “fake lion’s mane mycelium”, which we sent into the suspected dry lab. On the intake form, we told them we were expecting it to be a lion’s mane mycelium extract, standardized to 4% erinacines.

At this point we knew what we were up against, and we were determined to rectify the situation and give our fellow nootronauts a well deserved, genuine erinacine A containing lion’s mane mycelium product. Unfortunately, we were still naive enough at the time, to not realize what an incredibly complex project this would be!

For the next half decade or so, we got a constant barrage of questions and requests about lion’s mane mycelium, to the point that we drafted up a saved response that went out to thousands of eager nootropics enthusiasts.

Almost all of the Lion’s Mane mycelium out there, is grown on grains like brown rice. Since it is extremely difficult to separate mycelium from brown rice, the mycelium and brown rice are oftentimes just ground up together. This means that you are rarely getting a pure mycelium product. We are currently looking into novel cultivation methods so that we can offer a pure Mycelium product that is free from grains. Another roadblock for a Lion’s Mane mycelium product, is that it is currently not possible to test for Erinacine A. Since Erinacine A is the main compound of interest in a Lion’s Mane mycelium product, it is critical that we can actually test for it. We are actively working on developing testing methods for Erinacine A. Once we have established methods, and have figured out a reliable cultivation method, then we will start offering very high quality Lion’s Mane mycelium that will be unlike anything currently available! Until then, our Lion’s Mane fruiting body extracts are of very high quality and are very well liked by many of our customers. In fact, one of our best selling products is our fruiting body Lion’s Mane extracts!”

During that time frame, we worked tirelessly to solve the complex issues at hand. Everytime we solved one problem, a new problem would pop up that we needed to solve. Meanwhile, we continued to watch in disbelief as our competitors kept releasing clearly fraudulent products claiming an erinacine content, even though at this point we knew this was entirely impossible!

Now, more than 8 years later, we have finally done it, and we are proud to announce our ErinaMAX lion’s mane mycelium which has a real minimum erinacine A content of 0.5% (our first batch actually has 0.85% erinacine A!)

The Road Towards An Erinacine A Standard

During that time frame, we worked tirelessly to solve the complex issues at hand. Everytime we solved one problem, a new problem would pop up that we needed to solve. Meanwhile, we continued to watch in disbelief as our competitors kept releasing clearly fraudulent products claiming an erinacine content, even though at this point we knew this was entirely impossible!

The first major roadblock to solve, also proved to be one of the most difficult to solve. We had to produce our own erinacine A reference standard! At the time, our lab was a fraction of the size it is now, and we did not have the instruments nor skilled personnel required to pull this off. Thus, we started to expand the lab, and we brought in some incredibly talented scientists. We then needed some material to actually isolate erinacine A from. With all of the erinacine containing material reportedly on the market, you’d think this would be an easy task but surprise surprise, none of these commercially available lion’s mane mycelium products actually appeared to contain significant enough amounts of erinacine A. If there were products like this on the market, we could have simply bought large amounts of commercially existing erinacine A containing lion’s mane mycelium products to then extract and purify on our Buchi flash & prep HPLC instrument. Alas, we had to find different methods to pull this off. We worked together with a renowned lion’s mane mycelium researcher, and were able to acquire enough erinacine A containing lion’s mane mycelium from them in order to isolate and purify a few milligrams of erinace A. We sent some of this erinacine A out for third party NMR analysis to verify that we had indeed made an erinacine A reference standard, and we were successful! We then used the extremely small concentration of erinacine A reference standard to test our own fruiting body products for erinacine A, and a bunch of mycelium on grain lion’s mane products that were on the market. The results were clear, currently there wasn’t a product out there that had enough erinacine A in it, and cultivating lion’s mane mycelium on grains simply did not seem to produce enough erinacines. At this point however, we had consumed almost all of our reference standard but we had just enough to test a few more raw materials in order to find a source from which we could isolate greater quantities of erinacine A.

For a brief period of time I (Emiel, product specialist and author of this blog), produced a small amount of my own liquid culture lion’s mane mycelium. I cloned one of the lion’s mane fruiting bodies I was cultivating for the lab onto a sterile agar dish. Once this agar dish had become colonized with mycelium, I transferred some of this mycelium into a sterile nutrient broth, which had been optimized for erinacine A production. This broth was then stirred on a magnetic stir plate and allowed to cultivate for 14 days. I then took this liquid culture mycelium, and transferred all of it into a much larger vessel full of sterile nutrient broth (about 1 liter). I placed this vessel back on the lab stir plate, and allowed it to cultivate for 7 days. The liquid culture mycelium was then isolated, and via thin layer chromatography (TLC) we determined that this batch of liquid culture mycelium had indeed generated a significant quantity of erinacine A. However, when we tried to isolate pure erinacine A from it, we had far too little material to actually work with. We weren’t trying to make single digit milligram amounts of erinacine A reference standard at this point, we wanted hundreds of milligrams now!

However, this experiment did yield some valuable information, we could actually produce significant amounts of erinacine A in a liquid culture medium that was optimized for erinacine A production! To source more liquid culture lion’s mane mycelium, we had to find a partner to work with who specialized in liquid culture mycelium cultivation. You can imagine that if a liter of nutrient broth only yielded a very small amount of erinacine A, then you would need HUGE bioreactors to produce erinacine A containing mycelium at a commercial scale.

We finally found a partner who had also been doing research for years on liquid culture lion’s mane mycelium cultivation and had recently finally been successful in producing a batch with what they estimated to be about 0.5% erinacine A. We took the plunge and ordered 100 kilograms of this lion’s mane mycelium. It’s still mind-boggling to me to imagine how big these bioreactors are to produce that much mycelial biomass!

Once we received it at our facility, we did the same TLC test on it, and found that it indeed contained erinacine A. Now we had 100 kilos to play with, so we immediately started extracting massive amounts of it, to then further purify on our Buchi flash & prep HPLC instrument. Within a few months, we had over 250 milligrams of pure erinacine A! We used this reference standard to then determine exactly how much erinacine A was in the source material we acquired from our partners, and found that it contained a very significant amount of erinacine A, at 0.85%! They were right on point with their estimate of roughly 0.5% erinacine A, and working with them was such a pleasure that we decided we wanted to work together with them to bring out the first real commercially available erinacine A containing lion’s mane mycelium.

A New Era

So here we are, 8 years later in 2023, with what we believe to be the first genuine commercially available erinacine A containing liquid culture lion’s mane mycelium. For the first time ever, it is now possible to take a similar erinacine A lion’s mane mycelium which is featured in various animal and human studies. This does leave us with the question, why is erinacine A so important?

Erinacine A stimulates the biosynthesis of a neurotrophic factor called nerve growth factor (NGF). NGF interacts with the tropomyosin receptor kinase A (TrkA) receptor, a receptor pathway that is highly involved in neuroplasticity, neuroprotection and nerve regeneration. There aren’t a whole lot of naturally occurring compounds which can significantly upregulate the synthesis and release of NGF, and thus erinacine A has always held somewhat of a legendary status. Erinacine A also interacts with another neurotrophic factor called brain derived neurotrophic factor (BDNF). BDNF acts on a different receptor than NGF, namely the tropomyosin receptor kinase B (TrkB).

Similar to TrkA receptors, stimulation of TrkB receptors sets in motion many of the pathways responsible for the process of neuroplasticity. Furthermore, erinacine A also interacts with monoamine neurotransmitters, such as norepinephrine which plays a major role in memory consolidation, focus and mood. Taken together, it is clear to see why erinacine A has been held in such high regard, because it has an incredibly comprehensive effect on cognitive function and mood!

While ErinaMAX is currently only standardized for erinacine A, it will also contain other erinacines. We are still working hard on developing reference standards for the other erinacines, and in the future this will allow us to enhance the scope of our analytical chemistry. Having more erinacine reference standards on hand, will also allow us to perfect our extraction methodologies in order to create both full spectrum and targeted lion’s mane mycelium extracts! One of the other erinacines we are very interested in, is erinacine S. This erinacine has a very unique mechanism of action, and affects neuroplasticity via a pathway we had been previously unaware of. Erinacine S leads to the accumulation of two steroid hormones, pregnenolone and progesterone, in neural tissue. These two steroid hormones have a significant effect on neuroplasticity. Thus, by enhancing the levels of pregnenolone and progesterone in neural tissue, erinacine S produces a very unique neuroplasticity enhancing effect. Within the context of the NGF and BDNF modulating effects of erinacine A, it is clear to see why this novel effect of erinacine S is very exciting, lots of possible synergies!

Speaking of synergies, in our opinion one of the most interesting applications of Erinamax is in combination with our lion’s mane fruiting body offerings, especially our more nootropic focused lion’s mane 8:1 extract. This is due to the fact that the bioactives compounds within the lion’s mane fruiting bodies, such as the hericenones, can enhance the effects of NGF. Thus, combining hericenones with erinacine A which boost NGF synthesis, will yield the most comprehensive NGF focused effects.

Besides the cognitive benefits, Erinamax will also provide some very interesting effects for overall nerve health, specifically for nerve pain. Due to the fact that NGF is involved in the regeneration of damaged nerves, boosting NGF synthesis with Erinamax can have a beneficial effect on overall nerve function. This can lead to a pain management effect, especially in those who are experiencing nerve related pain. The pain management effects of Erinamax are likely to be further amplified by one of the other erinacines that should be in Erinamax, erinacine E. Erinacine E is a mild agonist at the kappa opioid receptor, which can produce robust pain management effects. The one downside with kappa opioid agonism, is that at high levels, it can induce a negative mood state. However, during our beta-testing we did not notice a negative mood state. In terms of kappa opioid agonism, Erinamax felt less pronounced than matrine which is a fairly significant kappa opioid agonist. Erinamax likewise also does not have as pronounced of a pain management effect when compared to matrine. However, in combination with its NGF modulating effects, Erinamax does have a very unique pain management effect for nerve related pain!

Subjectively, the effects of Erinamax are very different in comparison to our lion’s mane fruiting body offerings. The fruiting body offerings are more gentle, and take a little bit longer to produce their beneficial effects. With Erinamax, the effects are noticeable much more quickly. For example, on our Erinamax podcast Erika felt the effects of Erinamax kick in after about 30 minutes. On the other hand, she did not feel major acute effects with the lion’s mane 1:1 and 8:1. Within our beta-testing rounds, the acuteness of Erinamax also stood out for various participants. Now that Erinamax has been out for a week, we have also read a few reddit reports indicating rapid acute effects.

Even though ErinaMAX is the start of a new era of lion’s mane supplements, we also firmly believe that this is still the beginning. We have now cracked two very important aspects of lion’s mane mycelium. We have figured out how to cultivate high quality liquid culture lion’s mane mycelium in bioreactors, and the work we did on making an erinacine A reference standard is now going to allow us to produce other erinacine reference standards.

Better yet, our experience in creating an erinacine A reference standard is now also going to allow us to create reference standards for compounds present in the lion’s mane fruiting bodies, such as the hericenones. Together with our Mushroom Kingdom project, this is going to allow us to simultaneously innovate on both the mycelium and fruiting body sides. This collective innovation will allow us to make the most full spectrum lion’s mane products possible. It will also allow us to properly guide extraction methodology development. Currently, lion’s mane fruiting bodies and mycelium are being extracted with no way of knowing which of the beneficial compounds are getting concentrated, and which we are losing. With an arsenal of reference standards however, we will be able to develop advanced extraction strategies to create the best possible lion’s mane product!


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