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Method for improving yield of cordycepin obtained by liquid fermentation of cordyceps militaris
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A technology for fermentation of Cordyceps militaris and fungus, which is applied in the field of increasing the yield of cordycepin in liquid fermentation of Cordyceps militaris, and can solve the problems of easy pollution of feeding materials, inaccurate feeding, and low degree of increase in yield of cordycepin in Cordyceps militaris fungus fermentation liquid.
Active Publication Date: 2018-01-19
GANSU ACAD OF SCI INST OF BIOLOGY
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[0005] The present invention aims to provide a method for increasing the yield of cordycepin in the fermentation liquid of Cordyceps militaris, which overcomes the disadvantages of the prior art that the yield of cordycepin in the fermentation liquid of Cordyceps militaris is low, the feed is inaccurate, the feed is easy to pollute, and the feed process is complicated. technical flaw
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Embodiment 1
[0041] Embodiment 1; Step (1), Cordyceps militaris ( Cordyceps militaris ) CGMCC3.4655 was transferred to sterile PDA slant medium, cultured at 20°C in the dark for 6 days, and activated 3 times to obtain slant parent species.
[0042] Step (2), take a 3mm sized bacterial block from the mother species on the slant and inoculate it at the center of the culture medium on a sterile plate. The culture medium on the plate is: mix 33g of nutrientagar with 1000mL of pure water, pH7.3, in the dark at 20°C After culturing for 9 days, the mother species of the plate were prepared; the specific components of 33 g of nutrientagar were: 10 g of peptone, 3 g of beef powder, 5 g of sodiumchloride and 15 g of agar.
[0043] Step (3), use a puncher to inoculate 6 bacterial blocks with a size of 3mm on the same radius of the center of the mother species of the flat plate into a triangular flask of sterile seed medium (loading capacity: 300mL / 500mL), that is, the 6 bacterial blocks Inoculate...
Embodiment 2
[0052] Step (1), with embodiment 1;
[0053] Step (2), pH7.2; the same as in Example 1;
[0054] Step (3), with embodiment 1;
[0055] Step (4), with embodiment 1;
[0056] Step (5), the measured volume of the un-fed fermentation broth and the remaining amount of adenine were 95mL and 192.286µg / mL in turn, and substituted into the adenine feeding formula to calculate the mass concentration of the adenine feeding solution as 3.717 g / L .
[0057] Step (6) and step (7) are the same as in Example 1.
[0058] The test results showed that the measured value of the yield of cordycepin in the fermentation filtrate of the control group was 171.231 µg / mL, while the corrected value of the yield of the fermentation filtrate obtained in this example reached 893.798 µg / mL, which was 421.98% higher than that of the control.
Embodiment 3
[0060] Step (1), with embodiment 1;
[0061] Step (2), pH7.4; the same as in Example 1;
[0062] Step (3), with embodiment 1;
[0063] Step (4), with embodiment 1;
[0064] In step (5), the measured volume of the un-fed fermentation broth and the remaining amount of adenine were 115mL and 281.632µg / mL in turn, and the mass concentration of the adenine fed solution could be calculated as 3.976 g / L by substituting it into the adenine feeding formula .
[0065] Step (6) and step (7) are the same as in Example 1.
[0066] The test results showed that the measured value of cordycepin production in the fermentation filtrate of the control group was 138.601 μg / mL, while the corrected production value of the fermentation filtrate obtained in this example reached 856.207 μg / mL, which was 517.75% higher than that of the control group.
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Abstract
The invention provides a method for improving the yield of cordycepin in cordyceps militaris fermented liquid. The method comprises the following steps: transplanting a strain on a sterile PDA (PotatoDextrose Agar) slant culture medium, and culturing and activating to obtain a slant mother strain; transplanting the slant mother strain into a plate culture medium and culturing to obtain a flat mother strain; inoculating the flat mother strain into a seed culture medium and carrying out shaking table culture to obtain seed liquid; inoculating the seed liquid into an optimized culture medium andsealing a triangular flask opening by utilizing a sterile polypropylene bag; carrying out the shaking table culture to obtain fermented liquid to be supplemented; preparing an adenine supplementing solution according to an adenine supplementing formula; adding 100mL of the adenine supplementing solution into the fermented liquid to be supplemented to enable the content of adenine in the fermentedliquid to reach an optimized mass concentration, so as to obtain supplemented fermented liquid; after carrying out the shaking table culture, continually standing and culturing to obtain fermented liquid containing mycelia; filtering to remove the mycelia in the fermented liquid containing the mycelia, so as to obtain cordycepinfermentation filtrate. The method provided by the invention has theadvantages of simple technology and no pollution and the yield of the cordycepin in the cordyceps militaris fermented liquid reaches 956.020mug / mL.
Description
technical field [0001] The invention relates to the technical field of fungal fermentation, in particular to a method for increasing the yield of cordycepin in liquid fermentation of Cordyceps militaris. Background technique [0002] Cordycepin is 3' deoxyadenosine, which is a purinealkaloid with anti-virus, anti-inflammatory, tumor-inhibiting and improving immunity effects. In addition, cordycepin treats T lymphocytic leukemia through the ROS-mediated apoptosis pathway, and its research as a new drug for treating T lymphocytic leukemia has entered the second phase of clinical trials. Cordycepin is mainly produced by Cordyceps fungi. In addition, Aspergillus nidulans can also produce cordycepin, but Cordyceps militaris Cordyceps militaris The content of cordycepin in the fermented liquid is higher than that of other fungi. [0003] In recent years, there have been many studies on improving the production of cordycepin in the fermentation broth of Cordyceps militaris by te...
Claims
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