Lincomycin hydrochloride (LH) was purchased from Anhui Shuanghe Pharmaceutical Co., Ltd. (Anhui, China). ELISA kits were purchased from Shanghai MLBIO Biotechnology Co., Ltd. QIAamp DNA stool mini kit was manufactured by TIANGEN (Germany). Other reagents were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
According to our previous methods, CSP-1 was isolated from Cereus sinensis (19 (link), 21 (link)). After being soaked in NaCl solution (2%) to remove impurities, fresh Cereus sinensis was crushed and mixed with an equal volume of acetone for 12 h. The solution was filtered and freeze-dried to obtain degreased Cereus sinensis powder. The powder was mixed with distilled water at 72°C for 3 h and then precipitated with 3 times the volume of ethanol for 2 days. The above solution was centrifuged to obtain the precipitate, which was the crude polysaccharides. 50 mg/mL crude polysaccharide solution was mixed with 3% trichloroacetic acid for 12 h to remove protein. The solution was centrifuged, concentrated, dialyzed (3500D MWCO), and lyophilized. 5 mL of 20 mg/mL polysaccharide solution were further purified by column chromatography, involving DEAE-52 ion-exchange column and Sephadex G-100 column. 0.3 mol/L NaCl solution was used as eluate. The solution containing polysaccharides was collected, concentrated, dialyzed, and lyophilized to obtain CSP-1.
According to our previous methods, CSP-1 was isolated from Cereus sinensis (19 (link), 21 (link)). After being soaked in NaCl solution (2%) to remove impurities, fresh Cereus sinensis was crushed and mixed with an equal volume of acetone for 12 h. The solution was filtered and freeze-dried to obtain degreased Cereus sinensis powder. The powder was mixed with distilled water at 72°C for 3 h and then precipitated with 3 times the volume of ethanol for 2 days. The above solution was centrifuged to obtain the precipitate, which was the crude polysaccharides. 50 mg/mL crude polysaccharide solution was mixed with 3% trichloroacetic acid for 12 h to remove protein. The solution was centrifuged, concentrated, dialyzed (3500D MWCO), and lyophilized. 5 mL of 20 mg/mL polysaccharide solution were further purified by column chromatography, involving DEAE-52 ion-exchange column and Sephadex G-100 column. 0.3 mol/L NaCl solution was used as eluate. The solution containing polysaccharides was collected, concentrated, dialyzed, and lyophilized to obtain CSP-1.
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