Preparation method of cyclic dipeptide compound
A technology of compound and cyclic dipeptide, which is applied in the field of preparation of cyclic dipeptide, can solve the problem that the ability to synthesize active secondary metabolites is not fully exerted, and achieve the effect of simple composition, simple process, and low fermentation cost
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Embodiment 1
[0027] Embodiment 1: the selection of culture medium
[0028] Penicillium citrinum MNP12010101 was cultured under stress in high-concentration metal ion medium, the diversity of metabolites was analyzed by HPLC fingerprint method, and the antitumor activity of total fermented extract was detected by tetramethylazozole method (MTT method), so as to screen A metal ion can be added to the culture medium to promote the production of more anti-tumor active substances by Penicillium citrinum. Experimental metal ions include Mg 2+ , Mn 2+ , Fe 3+ 、Ba 2+ 、Co 2+ and Cu 2+ Wait. The results showed that Mg 2+ , Mn 2+ , Fe 3+ 、Ba 2+ and Cu 2+ When added to the fermentation medium, the diversity of metabolites of Penicillium citrinum was basically unchanged, and the antitumor activity of the total fermented extract was not significantly improved. Only the addition of 1.0g / L cobalt chloride (CoCl 2 ), the diversity of metabolites of Penicillium citrinum increases, and the antitu...
Embodiment 2
[0040] Embodiment 2: the selection of cobalt ion concentration of fermentation medium
[0041] The concentration of cobalt chloride in the fermentation medium was changed, and Penicillium citrinum MNP12010101 was cultured under stress. The diversity of metabolites was analyzed by HPLC fingerprint method, and the antitumor activity of total fermented extract was detected by tetramethylazozole method (MTT method). The specific experimental steps and methods are as follows:
[0042] (1) According to the method of Example 1, the seed liquid of Penicillium citrinum MNP12010101 was prepared;
[0043] (2) With the inoculation amount of step (2) seed liquid with 10% volume concentration, inoculate in the fermentation medium (250mL Erlenmeyer flask) that contains different concentrations of cobalt chloride of 50mL, see table 1 for cobalt chloride concentration, at 28 ℃, 200r / min shaking conditions, cultured for 5 days (see Table 1 for the dry weight yield of bacteria under different c...
Embodiment 3
[0051] Embodiment 3: the preparation of fermented total extract
[0052] Utilize the fermentation culture medium that contains the cobalt chloride of 1.0g / L, to Penicillium citrinum MNP12010101 large-scale fermentation culture, prepare fermentation total extract, concrete steps are:
[0053] Penicillium citrinum MNP12010101 was inoculated on the slant medium, and cultured at 28°C for 60 hours to obtain the activated strain spores; the activated strain spores were picked and inoculated into 100mL seed medium (500mL Erlenmeyer flask), Cultivate at 28°C and 200r / min shaking conditions for 36h to obtain seed liquid; transplant the seed liquid into 400mL fermentation medium (packed in a 1L Erlenmeyer flask) with an inoculum of 10% volume concentration, and inoculate at 28°C and 200r / min After culturing for 7 days under shaking conditions (the dry weight of the bacteria is 12.2 g / L), and then standing at 28° C. for 5 days, a culture was obtained. Following the steps above, 80 L of ...
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