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CN-121294151-B - Method for rapidly culturing ganoderma lucidum by liquid and application thereof

CN121294151BCN 121294151 BCN121294151 BCN 121294151BCN-121294151-B

Abstract

The invention discloses a method for rapidly culturing ganoderma lucidum by liquid and application thereof, wherein modified raw materials such as enzymolysis oat flour, sodium alginate-chitosan microcapsule microbial inoculum and the like are prepared; preparing a liquid culture medium composed of enzymolysis oat flour, deep sea fish peptone, modified soybean peptide, microcapsule microbial inoculum, lycium ruthenicum extract, sodium selenite and the like, then carrying out 'dark proliferation-blue light induction' two-stage directional fermentation, adding a methyl jasmonate inducer in a light stage in a pulse mode, and finally carrying out low-temperature after-ripening and microwave vacuum drying to obtain the product. The invention overcomes the defects of long culture period and low content of active ingredients in the traditional method, shortens the growth period to 9 days, ensures that the content of the ganoderma lucidum polysaccharide in the ganoderma lucidum product is not less than 25 percent, ensures that the beta-glucan accounts for more than 50 percent, and realizes the directional enrichment of selenium. The product can be used for preparing functional food for regulating immunity or auxiliary antitumor pharmaceutical composition, and realizes high-efficiency, standardized and functional production of Ganoderma.

Inventors

  • LIU BIN

Assignees

  • 河北岁神生物科技有限公司

Dates

Publication Date
20260512
Application Date
20250928

Claims (9)

  1. 1. A method for rapidly culturing ganoderma lucidum by liquid, which is characterized by comprising the following steps: (1) Prefabricating components of a modified culture medium: a. The preparation method of the enzymatic oat flour comprises preparing oat flour into 15-20% suspension, adding neutral protease 4000-6000U/g oat flour, hydrolyzing at 50-55deg.C and pH6.5-7.5 for 2-3 hr, inactivating enzyme, and spray drying to obtain enzymatic oat flour; b. The preparation of the sodium alginate-chitosan microcapsule microbial agent comprises the steps of mixing a ganoderma lucidum mycelium extract, a cordyceps militaris mycelium extract and a lactobacillus plantarum metabolite according to the weight ratio of (3-5) (2-4) (1) as a core material, dripping a 1.5% sodium alginate solution as a wall material into a 2% calcium chloride solution by adopting a sharp hole method to form gel beads, and coating and crosslinking for 10 minutes by using a 0.5% chitosan solution to prepare the microcapsule microbial agent; (2) Preparing a liquid culture medium, namely mixing, by weight, 10-20 parts of enzymolysis oat flour obtained in the step (1), 5-12 parts of deep sea fish peptone, 3-8 parts of modified soybean peptide with the molecular weight of less than 1000Da, 2-5 parts of microcapsule microbial inoculum obtained in the step (1) b, 1-3 parts of lycium ruthenicum extract, 0.01-0.05 part of sodium selenite and 150-250 parts of water; (3) The directional fermentation induction comprises sterilizing the culture of the step (2) for 15-20 min at 115-121 ℃, cooling to 25-28 ℃, inoculating 3-5wt% of ganoderma lucidum strain, firstly culturing for 3-4 days under the dark condition at 100-150rpm in a shaking way, then culturing for 3-5 days under the blue light irradiation condition in a shaking way at 80-120rpm continuously, and adding methyl jasmonate inducer once every 24 hours in the illumination stage in a pulse way to ensure that the final concentration of the methyl jasmonate inducer in the culture medium is 10-50 mu mol/L; (4) Harvesting and after-ripening, namely harvesting culture when the mycelium pellet density reaches 15-25g/L, filtering by a 100-mesh screen, collecting ganoderma lucidum mycelia, standing wet mycelia in a phosphate buffer containing 0.1% glutamine at 4 ℃ for after-ripening for 12-24 hours; (5) And directional drying, namely carrying out microwave vacuum drying on the mycelium after post-maturation, wherein the microwave power is 300-500W, the vacuum degree is-0.08 to-0.09 MPa, and the drying is carried out until the moisture content is less than or equal to 8%, so that the product is obtained.
  2. 2. The method according to claim 1, wherein in step (1) b, both the ganoderma lucidum mycelium extract and the cordyceps militaris mycelium extract are prepared by an ultrasonic-assisted water extraction method, and the polysaccharide content is not less than 30%.
  3. 3. The method of claim 1, wherein in step (2), the modified soy peptide is an oligopeptide component having a molecular weight of less than 500Da, which is separated by nanofiltration membrane after double-enzyme hydrolysis by flavourzyme and trypsin.
  4. 4. The method of claim 1, wherein in step (3), the methyl jasmonate inducer is added in the form of microcapsules, and the wall material is beta-cyclodextrin, so as to achieve a pulse-type slow release effect.
  5. 5. The method according to claim 1, wherein in the step (3), the blue light is irradiated at a wavelength of 450 to 470nm, the light intensity is 20 to 40. Mu. Mol. M-2. S-1, and the light-dark period is 12 hours light/12 hours dark.
  6. 6. The method according to claim 1, wherein in the step (4), the phosphate buffer solution has a concentration of 0.05m and a ph of 6.8, and the post-maturation process is performed in an anaerobic environment.
  7. 7. A meat ganoderma lucidum product produced by the method of any one of claims 1 to 6, characterized in that the meat ganoderma lucidum polysaccharide content is not less than 25% by dry weight, and wherein the beta-glucan component having an immunoregulatory activity is more than 50% by dry weight, and that the selenium element content is 50-200mg/kg by dry weight.
  8. 8. The ganoderma lucidum product according to claim 7, wherein the mycelium is hollow and porous, has a specific surface area increased by more than 100% as compared with a conventional liquid culture product, and is rich in terpenoid compounds induced by methyl jasmonate.
  9. 9. Use of the ganoderma lucidum product according to claim 7 in the preparation of functional food or health care products.

Description

Method for rapidly culturing ganoderma lucidum by liquid and application thereof Technical Field The invention belongs to the technical field of microbial culture and bioengineering, and particularly relates to a liquid submerged culture method of ganoderma lucidum. In particular to a method for realizing rapid, high-yield and high-quality culture of ganoderma lucidum and obtaining a functionalized product rich in specific active ingredients by innovating a culture medium formula and a directional fermentation induction technology and application thereof. Background Ganoderma lucidum, a rare large-scale colistin complex or specific fungus, is endowed with various health care functions in traditional cognition due to slow growth and peculiar morphology. Modern researches show that the compound is rich in bioactive substances such as polysaccharide, triterpene, protein and the like, has potential value in the aspects of regulating immunity, resisting oxidation and the like, and has increasingly growing market demands. However, the industrialization development of ganoderma lucidum is severely limited by the bottlenecks of low efficiency, uncontrollable quality and the like of the traditional culture mode. Currently, cultivation of ganoderma lucidum is mainly dependent on solid matrix cultivation or simple liquid stationary cultivation. The solid culture period is as long as several months to years, the occupied area is large, the labor is intensive, and the solid culture period is easy to be polluted by sundry fungus, so that the yield and the quality are extremely unstable. The existing liquid culture technology shortens the growth period to a certain extent, but has the obvious defects that firstly, the culture medium formula is mainly composed of glucose, peptone and other conventional raw materials, the nutrition components are single, the complex growth environment of the ganoderma lucidum in nature is difficult to simulate, the mycelium biomass accumulation is limited, the synthesis efficiency of key active components (such as beta-glucan) is low, secondly, the culture technology is extensive, the constant temperature oscillation at a single temperature is usually used, the precise regulation and control on different stages of growth and development of the ganoderma lucidum are lacking, the synthesis of high-value secondary metabolites cannot be effectively induced, thirdly, the harvesting treatment mode is simple, the high-temperature drying is often adopted, the degradation and inactivation of the thermosensitive active components are easy to cause, and the efficacy of the final product is influenced. The prior art has a plurality of limitations that limit the high-efficiency development and utilization of the ganoderma lucidum, namely, the traditional solid culture is like eating by the day, the period is too long, the success rate is low, and the requirement of large-scale production can not be met; secondly, although the speed is improved by simple liquid culture, the content of active ingredients of the obtained product is often inferior to that of a wild or solid culture, the efficacy is greatly reduced, the commercial value is limited, thirdly, the controllability of the whole culture process is poor, and a systematic bioengineering regulation and control means is lacked from inoculation, growth induction to harvest drying, so that the quality difference among batches is large, and the product standardization degree is low. In addition, the prior art rarely relates to directional nutrition fortification (such as selenium enrichment, selenium enrichment and other functional designs) of products through a culture process. Disclosure of Invention The invention aims to solve the core technical problems of overlong growth period, low content of bioactive substances, uncontrollable culture process, single function of the product and the like existing in the prior ganoderma lucidum culture technology. The traditional solid culture mode has low efficiency, and simple liquid culture is difficult to effectively induce and synthesize high-value secondary metabolites, so that the product quality cannot meet the high-end market demand. In order to solve the problems, the invention provides a method for rapidly culturing ganoderma lucidum based on an innovative culture medium formula and liquid induced by directional fermentation. The method has the core innovation that firstly, a liquid culture medium system with comprehensive nutrition and biological induction activity is constructed by introducing non-traditional raw materials such as enzymolysis modified oat flour, deep sea fish peptone, a special microencapsulated composite microbial inoculum and the like, and secondly, a two-stage directional fermentation process of 'dark proliferation-blue light induction' is adopted, and pulsed induction of methyl jasmonate is assisted, so that the growth and metabolic pathway of ganoderma lucidum are accurately regul