Psilocybe laccase with anti-tumor cell proliferation activity and preparation method thereof
A technology of Coprinus comatus and laccase, applied in the direction of antineoplastic drugs, medical preparations containing active ingredients, antiviral agents, etc., can solve the problems of toxic side effects and prominent drug resistance, and achieve HIV reverse transcriptase The effect of activity inhibition
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Embodiment 1
[0032] Embodiment 1, separation, purification and identification of Coprinus comatus laccase
[0033]The raw material of the present invention is to be isolated from the fruiting body of Coprinus comatus (Coprinus comatus) (China General Microorganisms Preservation Management Center, numbering CGMCC 5.615), utilizes PD medium (potato 200g, glucose 20g, add water to 1000ml), Obtain mycelium by fermentation under the following conditions: inoculate the Coprinus comatus species into a comprehensive PDA (200g potato, 20g glucose, 20g agar, add water to 1000ml) plate for activation, and cultivate at a constant temperature of 25°C until the mycelium is overgrown After plating, it was inoculated into PD medium, and after 7 days of shaking flask culture at 25°C and 160rpm, it was transferred to a liquid shake flask and cultured for 7 days to collect mycelium and store it in a -20°C refrigerator for use.
[0034] 1. Extraction and coarse separation
[0035] Suspend Coprinus comatus my...
Embodiment 2
[0057] Embodiment 2, the functional detection of Coprinus comatus laccase
[0058] 1. Functional detection of tumor cell proliferation inhibition in vitro
[0059] 1) Accurately weigh the pure laccase product obtained in Example 1, and prepare different doses of solutions (see Table 1) with serum-free culture medium to detect the activity of inhibiting tumor cell proliferation in vitro;
[0060] 2) Select human liver cancer cell line HepG2 (ATCC, HB-8065) and human breast cancer cell line MCF-7 (ATCC, HTB-22) as detection objects, and culture HepG2 and MCF-7 until the cells enter the logarithmic growth phase , the cells were arranged at a density of 8×10 3 Inoculate into a 96-well plate at a ratio of cells / well, and culture for 6 hours with serum-containing culture medium to make all the cells adhere to the wall;
[0061] 3) Remove the serum culture medium, wash the cells with PBS (Beijing Dingguo Changsheng Biotechnology Co., Ltd., BF-0013), replace with serum-free culture ...
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