A strain of Iberian monosporangium and its application
A technology for monosporic acid and cysts, which is applied to the application field of microorganisms and their natural products, can solve problems such as inability to obtain, and achieve the effects of low cost, simple and easy preparation method, and large extraction amount.
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Embodiment 1
[0034] Described monospora acid (Monospora acid) is made through the following steps:
[0035] (1) Strain activation: inoculate Monosporascus ibericus on a 90mm plate PDA medium, and culture at a constant temperature of 25°C for 72 hours to obtain activated strains;
[0036] (2) Fermentation culture: the activated bacterial classification prepared by step (1) is divided into 12 parts with a scalpel blade, and a part is inoculated in medium B (the preparation method of medium B: 60g rice and 100ml water are mixed, 121°C for 40 minutes), and the temperature is 25°C for static culture for 25 days to obtain the fermented product;
[0037] (3) Preparation of crude extract: Add 300ml of ethyl acetate to the fermented product prepared in step (2), cold-soak at 20-25°C for 12 hours, and filter with gauze to obtain ethyl acetate extract, which is repeatedly extracted 3 times , combining the three extracts to obtain the extract, and then recovering the ethyl acetate under reduced press...
Embodiment 2
[0047] Application of embodiment 2 monospora acid (Monospora acid) in the preparation of lipid-lowering drugs
[0048] Proprotein convertase subtilisin 9 (PCSK9) target assay
[0049] The specific method is as follows:
[0050] 2.1 Instrument reagents
[0051] The antibodies used in ELISA, such as His-tag primary antibody and HRP secondary antibody, were purchased from Abcam, and the EGF-AB protein and PCSK9 protein used in the experiment were all constructed, cultivated and purified by the Biocenter Laboratory. The remaining materials include 20xPBS (Sangon Biological), skimmed milk powder (BD), TMB high-efficiency chromogenic reagent (Solaibao Biological), 96-well plate for ELISA (Corning), concentrated sulfuric acid (Comeo), DPBS (HyClone), Tween 20 (Aladdin), ddH2O (Milli-Q preparation) and other necessary consumables.
[0052] The experimental instrument used Plate Reader EnVision (PerKinElmer) to read the OD value, and the constant temperature incubator (Shanghai Yihe...
Embodiment 3
[0073] The application of embodiment 3 monospora acid (Monospora acid) in the preparation anticoagulant medicine
[0074] anticoagulation test
[0075] The specific method is as follows:
[0076] 3.1 Experimental animals
[0077] Rex rabbits, male, weighing 2.0-2.5kg, were fed adaptively for one week. The temperature is 25±2°C, the light is 12 hours / day, the humidity is 40-45%, and there is free access to water and food.
[0078] 3.2 Instrument Reagents
[0079] HF6000-4 semi-automatic coagulation analyzer (Southern Hanfang Medical Instrument Co., Ltd.); TGL-16gR high-speed desktop refrigerated centrifuge (Shanghai Anting Scientific Instrument Factory); electronic balance (Mettle-Toledo, USA), sodium chloride injection ( Hebei Tiancheng Pharmaceutical Co., Ltd., A14091701); 0.109mol / L sodium citrate solution (self-made), vitamin K 1Injection (Tianjin Pharmaceutical Group Xinzheng Co., Ltd., 1403112); Breviscapine for injection (Hunan Hengsheng Pharmaceutical Co., Ltd., 20...
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