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I read an article a while back on how autoclaving renders spores inert. I'll have to attempt to find it again, but the gist was autoclaving at (120c?) for a set amount of time would simply destroy the spore wall and expose the internal proteins to outside influences where they could not survive and gestate.
Pressure and heat normally activate the enzymes responsible for germination of the sporoderm. Would probably be a good choice if heat doesnt break down the active ingredients. A low temperature tyndallization might be more appropriate so the actives are not decomposed and enzymatic activities continue to work.
In U.S. Pat. No. 6,316,002 the spores are soaked, germinated, then weakened with lytic enzymes or micronized physically. Finally it is dried and a methanol extraction is done and purified.
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The present invention provides a method for providing neuroprotection of dopaminergic neurons and treatment of Parkinson's Disease (PD). The method involves administering an effective amount of germination activated Ganoderma spore powders (GASP) into a mammal having PD.
Ganoderma (Ganoderma lucidum Leyss ex Fr. Karst) is a polyporous fungus which belongs to the class Basidiomycetes, the family Polypolaceae, and the genus Ganoderma. Ganoderma spores are tiny and mist-like spores of 5˜8 μm in sizes which have extremely hard and resilient, double-layer epispores, thus making them difficult to break open. The spores contain high concentrations of many bioactive substances, including, but are not limited to, polyunsaturated fatty acids, polysaccharides, vitamins, sterols, trace minerals, amino acids, and triterpenes. The GASP used in the present invention are sporoderm-broken (i.e., the double-layer epispores of the spores are broken so that the bioactive substances within the spores are released), which is produced by the method described in U.S. Pat. No. 6,316,002 (“the '002 patent). The entire content of the '002 patent is herein incorporated by reference.
Through the unique spore-breaking method described in the '002 patent, the bioactive substances within the GASP are recovered in high yields and the functional activities of the bioactive substances are successfully preserved.
As shown below is a general description of the method used in the '002 patent, which leads to the production of the GASP:
I. Soaking to induce germination: Mature and perfect spores of Ganoderma lucidum were carefully selected to carry out a soaking process to induce germination. Spores were kept in clear or distilled water, biological saline solution, or other nutritional solutions that could enable the spores of Ganoderma lucidum to germinate rapidly. Examples of nutritional solutions include coconut juice or a 1-5% malt extract solution, 0.5-25% extracts of Ganoderma lucidum sporocarps or Ganoderma lucidum capillitia, 0.1-5% of culture solution containing biotin, 0.1-3% of culture solution containing monobasic potassium phosphate and magnesium sulfate. The choice of solution would depend on the soaking time required, the amount of spores to be processed and other such factors as availability of materials. One or more of the above germination solutions could be used, with the amount added being 0.1-5 times the weight of the spores of Ganoderma lucidum. The soaking time can be determined according to the temperature of the water, and usually the soaking was carried out for 30 min to 8 h with the temperature of the water at 20-43° C. Preferably soaking times were 2-4 hours, and temperature of the water was 25-35° C.
II. Activation culture: The spores of Ganoderma lucidum were removed from the soaking solution and excess water was eliminated by allowing it to drip. The spores were then placed in a well-ventilated culturing box at a constant temperature and humidity so that spore activation culture could be carried out. The relative humidity of the culture was generally set at 65-98%, the culture temperature at 18-48° C. and the activation time lasted from 30 min to 24 h. Preferably humidity is 85-97% and temperature is 25-35° C. Using the method provided by the present invention, the activation of spores of Ganoderma lucidum reached a rate of more than 95%. During activation, the cell walls of the spores of red Ganoderma lucidum were clearly softened such that it was easier to penetrate the cell walls of the spores.
III. Treatment of the epispores: After the germination activation process, the spores were treated by enzymolysis. This process was carried out at a low temperature and under conditions such that enzyme activity was maintained, using chitinase, cellulase, or other enzymes, which are commonly used in the industry. The process was complete when the epispores lost their resilience and became brittle. Alternatively, physical treatments were carried out to penetrate the cell walls, for example, micronization, roll pressing, grinding, super high pressure microstream treatment, and other mechanical methods commonly used in the industry could be carried out, with a penetration rate of over 99%.
IV. Drying or extraction: Drying was carried out at low temperature using standard methods including freeze-drying or vacuum-drying etc., which are commonly used in the industry. The obtained product had a moisture content less than 4%. After drying, the bioactive substances were extracted by water or alcohol, or by thin film condensation. The extracted bioactive substances could be further purified by dialysis to ensure no contamination in the final products.
V. Pharmaceutical formulations of the bioactive substances: The bioactive substances can then be made into purified powders, extract pastes, solutions for injection, or for oral consumption. The invention also encompasses the manufacture of pharmaceutical preparations of the active substances, using well-known expedients and methods of manufacture known in the art. In addition, the bioactive substances can be dosed by any convenient method including tablets, capsules, solutions, suppositories, nasal sprays, paranterals, or injection devices. The choice of method for administration is determined by the clinical situation of the patient. The bioactive substances of the present invention, produced by the methods described, include active genes, inducers of the biotic potential promotor, inducers of the multicellular activator, inducers of interferon, lactone A, ganoderma polysaccharide, ganoderma spore fatty acids, ganoderma spore long chain alkyl hydrocarbon, ganoderma triterpenes, sterols, superoxide dismutase, vitamin E, active glycoprotein, certain growth factors, ganoderma acid A, superoxide dismutases (SOD), active glycoproteins, multiple active enzymes, and growth factors and so on. These bioactive substances, in a whole, contribute to the therapeutic uses described in the later sections.
GASP are non-toxic. The preferred method for administering GASP is through oral uptake. Currently, GASP are approved by the Food and Drug Administration (FDA) to be used as dietary supplement in the capsule form under the name of Enhanvol® and Holistol, sold by Enhan Technology Holdings International Company, Ltd. in Hong Kong. Each capsule of GASP contains 0.3 g of GASP. The recommended dosage of GASP, when used as dietary supplement, is 4 times every day, 4 capsules each time. Thus, for an adult of 60 kg, the daily dosage of GASP as dietary supplement is at about 0.08 g/kg of body weight per day.
By activating the spores the active ingredients are present in greater qualities as if they continue to synthesize as the spore germinate.
If you need lytic enzymes a.k.a. lysin mixtures PM Feelers, hes working on culturing protoplasts and is currently searching for a cheap source. He was looking particularly at trichoderma harzianum lytic enzymes from sigma aldrich which would be perfect for breaking the walls down. Perhaps a co-buy would benefit both of you and anyone else interested?
Heres a thread he recently started on the sourcing and use of lytic enzymes.
https://www.shroomery.org/forums/showflat.php/Number/22001124#22001124
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smegzilla69 
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yeah, if it involves a stove or any kind of solvent, it's SUPER-high tech.
YidakiMan
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