β-内酰胺酶抑制剂快速筛选方法的建立

发布时间：2018-03-13 07:23

  本文选题：耐药菌　切入点：β-内酰胺酶抑制剂　出处：《中国预防兽医学报》2017年09期 　论文类型：期刊论文

【摘要】：为建立用于抗耐药菌的β-内酰胺酶(β-lactamase)抑制剂快速筛选方法,本研究采用双层平板法,以2%的琼脂为底层,1%的含碘试剂为上层,以青霉素为β-内酰胺酶的底物,克拉维酸作为体系验证的阳性对照,通过比较透明圈形成大小,优化反应体系,并对190个微生物发酵产物进行了初步筛选。研究最终确定的筛选条件为,pH7.0的PBS缓冲液体系,底物青霉素(160万单位)浓度为50 mg/mL,β-内酰胺酶浓度为10 U/mL,40℃孵育40 min。通过测量反应体系透明圈形成大小来筛选具有活性的β-内酰胺酶抑制剂。结果证实该方法可以为耐药菌β-内酰胺酶抑制剂的筛选提供快速简单、易行、稳定、可靠的研究平台,并初步筛选获得了具有较高β-内酰胺酶抑制活性的化合物。
[Abstract]:In order to establish a rapid screening method of 尾 -lactamase inhibitors for resistant bacteria, a double-layer plate method with 2% Agar as the bottom layer and 1% iodine containing reagent as the top layer and penicillin as the substrate of 尾 -lactamase was studied. Clavulanic acid was used as a positive control to verify the system. By comparing the size of transparent circle, the reaction system was optimized, and 190 microbial fermentation products were screened. The final screening conditions were determined as PBS buffer system with pH 7.0. The concentration of penicillin (1.6 million units) was 50 mg / mL, and the concentration of 尾 -lactamase was 10 U / mL at 40 鈩，

本文编号：1605361