Mass production of a S-layer protein of Bacillus thuringiensis and its toxicity to the cattle tick Rhipicephalus microplus
Mass production of a S-layer protein of Bacillus thuringiensis and its toxicity to the cattle tick Rhipicephalus microplus
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DOI：
10.1038/s41598-019-53854-3
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发表时间：
2019-11-26
影响因子：
4.6
通讯作者：
Pena-Chora, Guadalupe
中科院分区：
文献类型：
--
作者：
Lormendez, Caleb C.;Fernandez-Ruvalcaba, Manuel;Pena-Chora, Guadalupe
最常用的用于控制农业、森林和人类疾病媒介昆虫的生物杀虫剂来自苏云金芽孢杆菌，它在孢子形成阶段开始产生Cry和Cyt杀虫蛋白。一些苏云金芽孢杆菌菌株还产生对某些害虫有毒的S层蛋白。S层蛋白是细菌和古菌中含量最丰富的蛋白质。与Cry和Cyt不同，这种蛋白质设计高性能大规模生产的关键特征是它们在营养阶段持续表达，而Cry和Cyt仅限于产孢期。本工作采用分批培养发酵技术，对苏云金芽胞杆菌GP543菌株表达的对牛微立克蚤有毒力的S层蛋白进行了批量生产。此外，孢子-蛋白质复合体在300微克·毫升(-1)的剂量下，14d后对微小核桃蛾雌成虫的死亡率为75%。致死浓度50为69.7微克·毫升(-1)。200mU·g·L~(-1)的孢子蛋白复合体还可使产卵质量下降13%，孵化抑制率由80%降至92%。因此，这可能是控制这种寄生虫的一个很好的选择。S层蛋白质合成的优势集中在生产新一代蛋白质在害虫防治上。这是首次报道大规模生产对毒性负责的S层蛋白。
The most commonly used biopesticides to control agricultural, forest and insect vectors of human diseases are derived from the bacterium Bacillus thuringiensis, which begins to produce Cry and Cyt insecticidal proteins during the onset of the sporulation phase. Some B. thuringiensis strains also produce S-layer proteins that are toxic to certain pests. S-layer proteins are the most abundant proteins in bacteria and archaea. This proteins' key trait to design high performace processes for mass production is their continuous expression during the vegetative phase, unlike Cry and Cyt, which are restricted to the sporulation phase. In this work, a S-layer protein expressed by the GP543 strain of B. thuringiensis that is toxic to the cattle tick Rhipicephalus microplus was mass produced using the batch culture fermentation technique. In addition, the spore-protein complex showed a mortality rate of 75% with a dose of 300 mu g-mL(-1) on adult females of R. microplus after fourteen days. The lethal concentration 50 was 69.7 mu g-mL(-1). The treatment also caused a decrease of 13% in the weight of the mass of oviposited eggs with 200 mu g-mL(-1) of the spore-protein complex and inhibition of the hatching of eggs from 80 to 92%. Therefore, this could be a good option for controlling this parasite. The advantages of S-layer protein synthesis are focused on the production of a new generation of proteins in pest control. This is the first report on the mass production of an S-layer protein that is responsible for toxicity.