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Cordyceps militaris mutant strain with high yield of cordycepin and its application
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A technology of cordycepin and Cordyceps militaris, applied in the field of agricultural microorganisms, can solve the problems of strain degeneration, industrial development, economic loss, poor quality, etc.
Active Publication Date: 2020-08-21
北京市辐射中心
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[0004] In recent years, the scale of artificial cultivation of Cordyceps militaris has continued to expand, but the degradation of the strain has always been a difficult problem restricting the development of the industry. The main performance is that it does not grow or only grows a very small number of fruit bodies, and the quality is poor, which will cause great economic losses.
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Embodiment 1
[0026] Example 1 Heavy carbon ion beam implantation mutagenesis breeding of Cordyceps militaris mutants with high yield of cordycepin
[0027] 1. Strains
[0028] The starting strain: Cordyceps militaris (Cordyceps militaris) No. CM-2, preserved by the Microbiological Laboratory of Beijing Radiation Center.
[0029] 2. Media and reagents
[0030] (1) culture medium
[0031] PDA medium: PDA powder (purchased from BD Company) 39g, distilled water to 1000mL, pH natural, sterilized at 121°C for 20min.
[0032] Liquid medium, seed liquid medium: 30g sucrose, 30g peptone, 1g potassium dihydrogen phosphate, 0.5g magnesium sulfate, with natural pH, heated to dissolve, distilled water to 1000mL, sterilized at 115°C for 30min.
[0033] Cordyceps militaris solid medium: 60g rice, add 60mL nutrient solution, and sterilize at 115°C for 30min.
[0078] Embodiment 2 The solid culture of Cordyceps militaris mutant strain CGMCC No.14800
[0079] 1. Solid culture of mutant strains
[0080] The solid culture of the Cordyceps militaris strain uses liquid seeds, and the preparation method of the seed liquid is as described in Example 1 for liquid culture. Take the cultured 5mL seed solution and evenly sprinkle it on the surface of the sterilized and cooled solid medium. Cultivate at a constant temperature of 25°C and avoid light. The surface of the medium is covered with mycelia for 7 days. The mycelium and rice grains on the upper layer of the medium are broken up into small particles with sterile tweezers in an ultra-clean bench, and spread flat in the culture bottle. Then move into the artificial climate incubator at 20-23°C, set daytime for 8h, humidity 80%, temperature at 23°C, light intensity at 1200lux, night at night for 16h, humidity at 75%, and temperature at 22°C. Yellow becomes orange, and yellow gradually deep...
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Abstract
The invention relates to a mutant strain of cordyceps militaris (Cordyceps militaris) with high yield of cordycepin and its application. A mutant strain of Cordyceps militaris CGMCC No.14800 with high yield of cordycepin and fruiting bodies obtained by heavy carbon ion beam implantation mutagenesis technology was used as a fermentation strain for liquid fermentation to prepare cordycepin. Cordycepin production can reach up to 929.390μg / mL, which is 8.15 times of the yield of cordycepin in the fermentation broth obtained from the starting strain; for the solid culture of the strain CGMCC No.14800, the fruiting body yield can reach up to 22.376g / bottle, which is the highest yield of the starting strain. 1.72 times of the yield; the fruiting body cordycepin production can reach up to 11777.094μg / g, which is 1.75 times the fruiting body cordycepin production of the starting strain. The invention provides a new channel for the development of the Cordyceps militaris strain with high yield of cordycepin.
Description
technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a mutant strain of Cordyceps militaris with high yield of cordycepin and its application. Background technique [0002] Cordyceps militaris (Cordyceps militaris), also known as Cordyceps militaris, is a model species of the genus Cordyceps. hotspot. [0003] Cordycepin is the most important bioactive component of Cordyceps militaris, which has antitumor, immune regulation, antibacterial, antiviral, and anti-inflammatory effects. Currently, the price of cordycepin on the market is expensive, mainly due to the limitation of conditions that can only be chemically synthesized in the laboratory, and large-scale production cannot be realized. The existing cordycepin on the market is mainly extracted and isolated from the culture of Cordyceps militaris. Therefore, the breeding of Cordycepin militaris strains with high yield plays an important role in re...
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