Lion’s Mane Extract Made with Ultrasonics
Extracts from the fungus species Hericium erinaceus, known as lion’s mane mushroom, are most efficiently produced using ultrasonication. Ultrasonic extractors rapidly break open the fungal cell matrix and allow for the complete extraction of bioactive compounds from the lion’s mane mycelium and fruiting body.
Ultrasound-Assisted Lion’s Mane Mushroom Extraction
Bioactive Compounds in Lion’s Mane: Hericium erinaceus, also known under the common names of lion’s mane, Japanese yamabushitake, pom pom, bearded tooth, hedgehog or monkey head mushroom, is a fungus that is used since decades as a traditional medicine and therapeutic. Lion’s mane contains many bioactive compounds polysaccharides, sterols, glycoproteins, terpenoids (e.g. erinacines), as well as phenolic and volatile compounds (e.g. hericenones). These substances are known for their antioxidative, antidiabetic, anticancer, anti-inflammatory, antimicrobial, anti-hyperglycemic, and hypolipidemic effects. Scientific research has shown that lion’s mane compounds can improve neuronal development and function and might protect nerves from becoming damaged. Therefore, it is currently tested as a therapeutic for dementia.
Ultrasonic Lion’s Mane Extraction:
Ultrasonic extraction of lion’s mane is a technique, which applies high-power ultrasound in order to extract bioactive compounds from the Lion’s Mane mushroom (Hericium erinaceus) fruiting body or mycelium. The Lion’s Mane mushroom is a well-known medicinal mushroom, and it contains various health-promoting bioactive compounds such as polysaccharides, beta-glucans, hericenones, erinacines, and antioxidants.
The ultrasonic mushroom extraction process involves using probe-type ultrasonicators that create intense cavitation in a liquid medium (such as water, ethanol, or methanol) containing the mushroom material. The generated ultrasonic cavitation causes the cell walls of the mushroom material to break down, releasing the bioactive compounds into the liquid / solvent. The ultrasonic waves also enhance the mass transfer of the bioactive compounds from the mushroom material to the solvent, which increases the extraction efficiency.
Ultrasonic mushroom extraction is a very efficient and rapid isolation technique that does not require high temperatures or harmful chemicals. The extracted bioactive compounds can be used for various applications, such as dietary supplements, functional foods, and nutraceuticals. Furthermore, ultrasonic lion’s mane extraction method is environmentally-friendly and sustainable, making it an ideal choice for extracting bioactive compounds from natural sources.
- high efficiency
- purely mechanical extraction effects, which makes the extraction gentle
- simple operation
- very short processing time
These advantages make sonication an excellent extraction technique for high-quality mushroom extracts and are the reason, why Hielscher ultrasonicators are used worldwide in laboratories and industry for the production of mushroom extracts.
Protocol for Ultrasonic Lion’s Mane Extraction
Valu et al. (2020) demonstrated a highly efficient extraction procedure for obtaining and concentrating the bioactive products of the H. erinaceus biomass based on the principles of the ultrasonic extraction. The device used for extraction was a Hielscher ultrasonic processor (Hielscher UIP1000hdT, 1000 Watts, 20 kHz) with sonotrode BS4d40 (40 mm diameter). Before the extraction experiments, the ultrasonic processor was calibrated to determine the net power consumption. During the sonication process, this value was automatically deducted from the gross energy consumption, thus allowing the net power delivered to the extraction medium to be found. During the experiments, the samples were placed in an ice bag with continuous magnetic stirring to maintain a low sample temperature. After completion of the extraction, the samples were vacuum filtered and then centrifuged (2500× g for 5 min). A rotary evaporator was used for water and alcohol elimination from the supernatants. The remaining water and alcohol residues from the samples were subjected to lyophilization in order to obtain a powder extract. Alternatively, the solvent can be removed using a vacuum filter and a rotary vacuum evaporator in order to get a mushroom concentrate.
The optimized extraction conditions using ultrasound were the following:
- ultrasonicator UIP1000hdT with sonotrode BS4d40: 100% amplitude, 100% cycle)
- dried, ground Hericium erinaceus
- solvent: 80% aqueous ethanol
- solvent–material ratio: 1:30 (g/mL)
- extraction time: 45 min
The total content of phenolics in this optimized H. erinaceus extract was 23.2 mg GAE/g DM, and in the DPPH test, the antioxidant activity reached an IC50 of 87.2 μg/mL.
The research team successfully demonstrated that ultrasonic extraction efficiently drives the isolation of antioxidants in Hericium erinaceus, particularly polyphenols and flavonoids correlated with the diterpenoid erinacine A, known for its high antioxidant activity.
(cf. Valu et al., 2020)
|Batch Volume||Flow Rate||Recommended Devices|
|1 to 500mL||10 to 200mL/min||UP100H|
|10 to 2000mL||20 to 400mL/min||UP200Ht, UP400St|
|0.1 to 10L||0.1 to 2L/min||UIP1000hdT|
|0.1 to 20L||0.2 to 4L/min||UIP2000hdT|
|10 to 100L||2 to 10L/min||UIP4000hdT|
|15 to 150L||3 to 15L/min||UIP6000hdT|
|n.a.||10 to 100L/min||UIP16000|
|n.a.||larger||cluster of UIP16000|
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Literature / References
- Valu, Mihai-Vlad; Liliana Cristina Soare; Nicoleta Anca Sutan; Catalin Ducu; Sorin Moga; Lucian Hritcu; Razvan Stefan Boiangiu; Simone Carradori (2020): Optimization of Ultrasonic Extraction to Obtain Erinacine A and Polyphenols with Antioxidant Activity from the Fungal Biomass of Hericium erinaceus. Foods 9, No. 12, 2020.
- Valu, M.-V.; Soare,L.C.; Ducu, C.; Moga, S.; Negrea, D.; Vamanu, E.; Balseanu, T.-A.; Carradori, S.; Hritcu, L.; Boiangiu, R.S. (2021): Hericium erinaceus (Bull.) Pers. Ethanolic Extract with Antioxidant Properties on Scopolamine-Induced Memory Deficits in a Zebrafish Model of Cognitive Impairment. Journal of Fungi 2021, 7, 477.
- Venturella, G.; Ferraro, V.; Cirlincione, F.; Gargano, M. L. (2021): Medicinal Mushrooms: Bioactive Compounds, Use, and Clinical Trials. International Journal of Molecular Sciences, 22(2), 634.
- Picture of Hericium By Jim Champion / Hericium erinaceum on an old tree in Shave Wood, New Forest / CC BY-SA 2.0
Facts Worth Knowing
Bioactive Mushroom Compounds from Mycelium vs Fruiting Body
Both mycelium and fruiting body extracts can be produced with ultrasonic extraction and both have their own unique benefits. Which one is better depends on the specific use case and desired outcomes.
Mycelium extracts are generally less expensive and easier to produce in large quantities than fruiting body extracts, making them more accessible. Mycelium also contains many beneficial compounds such as polysaccharides, ergosterol, and enzymes.
On the other hand, fruiting body extracts contain higher levels of beta-glucans, triterpenoids, and other compounds that have been linked to health benefits. Fruiting bodies also tend to have a more diverse range of compounds and may be more potent in some cases.
Ultimately, the choice between mycelium and fruiting body extracts will depend on the specific application and desired effects. If you’re looking for immune support, for example, a mycelium extract may be a good option due to its high polysaccharide content. If you’re looking for cognitive support, a fruiting body extract may be a better choice due to its high triterpenoid content. It’s also worth noting that high-quality extracts from both mycelium and fruiting body sources can be effective and beneficial for a variety of purposes.
Bioactive Compounds in Lion’ Mane
Very important and well-studied bioactive metabolites also include the erinacines (A-I), a group of cyathin diterpenoids extracted from the mycelium of Hericium erinaceus or lion’s mane or yamabushitake, and hericenones (C-H), benzyl alcohol derivatives extracted from the fruiting body. Both groups of compounds can easily pass through the blood–brain barrier and have demonstrated neurotropic and neuroprotective effects. They are reported to induce nerve growth factor (NGF) synthesis, both in vitro and in vivo. However, this medicinal mushroom also has antioxidative, anti-inflammatory, anticancer, immunostimulant, antidiabetic, antimicrobial, hypolipidemic, and antihyperglycemic properties, although its most frequent use is for the treatment of neurodegenerative diseases and cognitive impairment.
Erinacin A, the main representative of the erinacine group, has been proven to have an effective protective effect against Parkinson’s disease. In a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of Parkinson’s disease, erinacin A produced a reduction of MPTP-induced dopaminergic cell loss, apoptotic cell death induced by oxidative stress, and the levels of glutathione, nitrotyrosine, and 4-hydroxy-2-nonenal (4-HNE); it also reversed MPTP-associated motor deficits, and reduced the impairment of 1-methyl-4-phenylpyridinium (MPP)-induced neuronal cell cytotoxicity and apoptosis, through an endoplasmic reticulum (ER) stress-sustained activation of the IRE1α/TRAF2, JNK1/2, and p38 MAPK pathways, the expression of C/EBP homologous protein (CHOP), IKB-β, and NF-κB, as well as Fas and Bax. This metabolite was also found to be effective against ischemic stroke, as reported in a study on rats in which the reduction of neuronal apoptosis, as well as the size of the stroke cavity in the brain by targeting iNOS/reactive nitrogen species (RNS) and p38 mitogen-activated protein kinase (MAPK)/CCAAT enhancer-binding protein homologous protein (CHOP) pathways, was observed.
Erinacin A was also reported to have significant antitumor activity in human gastric cancer TSGH 9201 cells, in which it induced significant apoptosis associated with increased phosphorylation of focal adhesion kinase/protein kinase FAK/Akt/p70S6K and serine/threonine kinase PAK-1 pathways. It also resulted in increased cytotoxicity and ROS generation, the reduced invasiveness and activation of caspases, and the expression of tumor necrosis receptor TRAIL. The strong antitumor action of this metabolite was subsequently confirmed by a recent study both in vitro in two human colon cancer cell lines (DLD-1 and HCT-116) and in vivo in a mouse model that further clarified its mechanisms. Treatment effects included stimulation of the extrinsic apoptosis activation pathways (TNFR, Fas, FasL, caspases), suppression of the expression of the antiapoptotic molecules Bcl-2 and Bcl-XL, and phosphorylation of Jun N-terminal kinase JNK1/2, responsive to stress stimuli, NF-κB p50 and p330. It was also demonstrated that the upregulation of death receptor molecules through the JNK MAPK/p300/NF-κB pathway is mediated by the modification of histone H3K9K14ac; the results of the in vivo assay revealed, in fact, increased levels of histone H3K9K14ac, as well as histone acetylation on Fas, FasL, and TNFR promoters.
Another erinacin, erinacin C, is known for its antineuroinflammatory and neuroprotective actions, which could be achieved through a mechanism of inhibition of IκB, p-IκBα (involved in the upstream NF-κB signal transduction cascade), and inducible nitric oxide synthase (iNOS) protein expression, and the activation of the Nrf2/HO-1 stress-protective pathway. The treatment of human BV2 microglial cells with LPS-induced inflammation resulted in reduced levels of nitric oxide (NO), IL-6, TNF-α, and iNOS, the inhibition of NF-κB expression, and the phosphorylation of IκBα (p-IκBα) proteins, as well as the inhibition of Kelch-like ECH-associated protein 1 (Keap1), and increased nuclear transcription factor erythroid 2-related factor (Nrf2) and the expression of the heme oxygenase-1 (HO-1) protein.
(excerpt from Venturella et al., 2021)