多位点生物杀虫毒素BtA形成的HPLC分析

 基于多位点生物杀虫毒素理论和生物耦合技术，研制生物杀虫毒素BtA，为新生物农药的开发提供了一种新思路、新方法和新手段。将苏云金芽孢杆菌（B.t.）晶体进行酶解改造，形成带末端氨基的原毒素；将阿维菌素的羟基进行激活、衍生化，形成带羧基的阿维菌素衍生物（Abamectin-COONa）；再利用氨基-羧基偶联剂（EDC）进行两种生物毒素的生物耦合。利用反相液相色谱（HPLC）检测不同反应时间的BtA生物耦合体系，以确定生物耦合反应的发生；通过反应底物两两组合的分析比较，识别生物耦合产物BtA生成的色谱特征，分析生物耦合产物---多位点生物杀虫毒素BtA的产生过程。

HPLC Analysis on Formation of the Multi-Bioinsecticidal-Toxin BtA

Based on the theory of multi-bioinsecticidal toxin and the technique of bioconjugation,The multi-bioinsecticidal-toxin BtA was worked out in order to provide a new idea and a new method for development of a new type of biocide. The bioconjugation procedure of BtA included: decomposing Bacillus thuringiensis (B.t.) crystal by DTT into the protoxin with an end amino group; activation and derivation of abamectin to form abamectin-COONa with a carboxylic group; final bioconjugation by using amino-carboxylic conjugator EDC to achieve the substance combination of two biotoxins. In the detecting program,the experiments were carried out with HPLC (High Pressure Liquid Chromatogram) on measuring BtA bioconjugation system at different reaction times to identify the occurrence of bioconjugation reaction,on comparing BtA bioconjugate system with the three reactants and three two-reactant combinations to distinguish bioconjugated product BtA,and on a qualitative analysis of the bioconjugated product BtA. The results proved that a new bioconjugated substance was confirmed to present in BtA bioconjugation system,which was named as multi-bioinsecticidal-toxin BtA.