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

Lion’s Mane fruiting body powders and extracts were already firmly on our menu. During this period, the r/nootropics subreddit was vibrant, fostering discussions and research on 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.

Erinacine A generated buzz, with Redditors and marketers hyping its cognitive benefits from 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! Analytical testing complexity means you can’t insert mycelium and get precise erinacine A measurements automatically. 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.

Analyzing Erinacine A Content

Run lion’s mane mycelium sample through UPLC; if erinacine A present, it will show on chromatogram. Analyzing lion’s mane mycelium for erinacine A content requires a reference standard for accurate measurement. Competitors’ assertions of 4% erinacines in their lion’s mane mycelium products puzzled us greatly. We were oblivious to widespread testing fraud in the supplement industry, delaying our realization of competitors’ claims.

Detection of Testing Fraud

Nearly a year of research uncovered widespread use of fraudulent dry labs in the industry. To confirm suspicions, we sent a spoofed “lion’s mane mycelium” sample to a suspected dry lab for testing. Using functional yeast extract, we simulated lion’s mane mycelium by adding fake beta-glucans and spiking it 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 specified it as a lion’s mane mycelium extract, standardized to 4% erinacines.

Addressing Industry Challenges

We were determined to provide genuine erinacine A containing lion’s mane mycelium for our fellow nootronauts. Unfortunately, we were still naive enough at the time, to not realize what an incredibly complex project this would be! We fielded constant inquiries about lion’s mane mycelium, necessitating a templated response for the multitude of eager nootropics enthusiasts.

Cultivation and Testing Challenges

Almost all of the Lion’s Mane mycelium out there, is grown on grains like brown rice. Separating mycelium from brown rice proves daunting, often resulting in a mixture of both when ground. This means that you are rarely getting a pure mycelium product. Researching new cultivation techniques to provide grain-free, pure mycelium products is underway. Another roadblock for a Lion’s Mane mycelium product, is that it is currently not possible to test for Erinacine A. Testing for Erinacine A is crucial due to its significance in Lion’s Mane mycelium products.

Future Developments and Quality Assurance

We are actively working on developing testing methods for Erinacine A. Once we have established methods and figured out a reliable cultivation method, we will start offering very high-quality Lion’s Mane mycelium. This product will be unlike anything currently available! Until then, our Lion’s Mane fruiting body extracts remain of high quality, favored by many customers. In fact, one of our best selling products is our fruiting body Lion’s Mane extracts!”

Persistent Problem Solving

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, competitors released fraudulent products, falsely claiming erinacine content, though we knew it was impossible. Now, over 8 years later, we proudly announce ErinaMAX lion’s mane mycelium, with a minimum 0.5% erinacine A content (initial batch: 0.85%).

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, competitors persisted in releasing fraudulent products, falsely claiming erinacine content, despite our awareness of its impossibility.

Sourcing Erinacine A Reference Standard

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 that time, our lab was much smaller, lacking the necessary instruments and skilled personnel for the task. 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.

Isolating 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. Then, we could extract and purify them on our Buchi flash & prep HPLC instrument.

Collaboration and Experimentation

Alas, we had to find different methods to pull this off. We collaborated with a respected lion’s mane mycelium researcher to obtain erinacine A-containing lion’s mane mycelium. This allowed us to isolate and purify a few milligrams of erinacine A. We sent some of this erinacine A for third-party NMR analysis to verify our erinacine A reference standard, and we succeeded! The results were clear: currently, no product had enough erinacine A, and cultivating lion’s mane mycelium on grains didn’t yield sufficient erinacines. At this point, however, we had almost depleted our reference standard. We had enough left to test additional raw materials to isolate more erinacine A.

Liquid Culture Experimentation

For a brief period, I (Emiel, product specialist and blog author), cultivated 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 the agar dish was colonized, I transferred some mycelium into a nutrient broth 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.

Scaling Up Production

The liquid culture mycelium was isolated, and thin layer chromatography (TLC) confirmed it had generated a significant quantity of erinacine A. When attempting to isolate pure erinacine A, we found insufficient material to work with effectively. We weren’t aiming for single digit milligram amounts of erinacine A reference standard anymore; we needed hundreds of milligrams.

Partnering for Success

However, this experiment provided valuable information: we could produce significant amounts of erinacine A in an optimized liquid culture medium! To acquire more liquid culture lion’s mane mycelium, we partnered with specialists in liquid culture mycelium cultivation. If a liter of broth yields a small amount, large bioreactors are needed for commercial production.

Successful Collaboration

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!

Production and Validation

Upon arrival at our facility, we conducted the same TLC test and confirmed its erinacine A content. With 100 kilos in hand, we began extracting large quantities, ready for further purification using 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 and NGF Stimulation

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).

Neuroplasticity and Monoamine Neurotransmitters

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.

Future Development and Erinacine S

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!

Synergistic Effects with Lion’s Mane

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.

Nerve Health and Pain Management

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!

User Experience and Product Innovation

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.

Advancements in Lion’s Mane Supplements

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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