Somers P J, Yao R C, Doolin L E, McGowan M J, Fukuda D S, Mynderse J S
Eli Lilly and Company, Lilly Corporate Center, Indianapolis, Indiana 46285.
J Antibiot (Tokyo). 1987 Dec;40(12):1751-6. doi: 10.7164/antibiotics.40.1751.
A new method of screening for chitinase inhibitors in crude fermentation broths as a means of discovering new insecticidal leads has been developed. In this procedure soluble Remazol brilliant violet 5R dye-coupled chitin degradation products released from insoluble chitin azure substrate by hydrolysis with Streptomyces griseus chitinase are filtered in 0.45 micron Millititer HA 96 well filtration plates and collected in 96 well microtiter plates. Inhibitors of this reaction are detected by a decrease in absorbance (570 nm) of the filtrate. A chitinase inhibitor, designated A82516, produced by culture A82516 was discovered using this screen. Purified A82516 was found to have an IC50 of 3.7 X 10(-6) M for S. griseus chitinase. At a test concentration of 0.27 mg/ml, A82516 was 100% effective in preventing development of house fly larvae to pupae. Allosamidin, a recently reported chitinase inhibitor in vitro, has spectral properties identical to A82516.
已开发出一种在粗发酵液中筛选几丁质酶抑制剂的新方法，以此作为发现新的杀虫先导化合物的手段。在此过程中，用灰色链霉菌几丁质酶水解不溶性几丁质天蓝底物释放出的可溶性活性艳紫5R染料偶联几丁质降解产物，在0.45微米的Millititer HA 96孔过滤板中过滤，并收集于96孔微量滴定板中。通过滤液吸光度（570纳米）的降低来检测该反应的抑制剂。利用此筛选方法发现了由培养物A82516产生的一种几丁质酶抑制剂，命名为A82516。纯化后的A82516对灰色链霉菌几丁质酶的IC50为3.7×10^(-6) M。在0.27毫克/毫升的测试浓度下，A82516在阻止家蝇幼虫发育成蛹方面的效果达100%。Allosamidin是最近报道的一种体外几丁质酶抑制剂，其光谱特性与A82516相同。
