Use of pachycladous algal onychin
A technology of phycocyanin and thick branch, applied in the field of medicine, can solve the problems such as the same structure and activity that are not seen
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Embodiment 1
[0028] Embodiment one: the preparation of compound Crassicaulisine (Crassicaulisine)
[0029] (1) Extraction: soak 4.6 (kg) of air-dried Chondria crassicaulis Harv. in industrial methanol at room temperature and extract completely. The extract was concentrated under reduced pressure to obtain methanol extract. Dissolve the methanol extract in water to form a suspension with CH 2 Cl 2 Extract, concentrate the extract to get CH 2 Cl 2 Extract (70g).
[0030] (2) Separation: CH 2 Cl 2 The extract was subjected to repeated silica gel column chromatography, and the elution gradient was gradually increased with petroleum ether-ethyl acetate-methanol, and the fractions eluted with MeOH:EtOAc=1:4 were collected, then applied to the silica gel column, and used CHCl 3 :MeOH=9:1, the compound Crassicaulisine (538 mg) was obtained.
Embodiment 2
[0031] Example 2: PTP1B inhibitory activity experiment of the compound Crassicaulisine
[0032] experimental method:
[0033] The protein tyrosine phosphatase PTP1B gene used for screening was constructed on the pGEX-KG vector by the method of molecular cloning, and then transformed into E. coli expression strain BL21(DE3), induced by IPTG and broken through the glutathione agarose affinity layer The GST fusion protein was obtained by column purification. After separation and identification by 12% SDS-PAGE gel, the purity reached above 90%. Further analysis of its enzymatic properties (K m 、k cat ) are consistent with the recombinant protein reported in the literature.
[0034] Using the ultraviolet substrate pNPP (test principle in the summary of the invention), the product obtained by hydrolyzing the phospholipid bond in the substrate pNPP by PTP1B has a strong light absorption at 410nm, and the change of light absorption at 410nm can be directly monitored to observe the...
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