Department of Biological Functions Engineering, Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, Kitakyushu Science and Research Park, Kitakyushu, Fukuoka, Japan.
Bioindustry Division, Kyushu Medical Co, LTD, Fukuoka, Japan.
FEBS Open Bio. 2022 Aug;12(8):1534-1541. doi: 10.1002/2211-5463.13448. Epub 2022 Jun 15.
Cry toxins from Bacillus thuringiensis (Bt) have been extensively applied in agriculture to substitute the use of chemical insecticides. We have previously reported the use of a coexpression system in which late embryogenesis abundant (LEA) peptides under the control of the lac promoter increase the expression of insecticidal proteins in Bt. The use of lactose to induce the expression of LEA peptides may be a desirable alternative to isopropyl β-D-thiogalactopyranoside, the most frequently used inducer for recombinant protein expression. In this study we investigated the use of lactose as an inducer for optimal protein expression. We observed enhanced insecticidal Cry protein expression by applying a simple technique based on intermittent induction, and then optimized concentration and the point of induction time from the 11 h to the 15 h. Our data suggest that intermittent induction of lactose might be a new technique for the enhancement of bacterial protein expression.
苏云金芽孢杆菌的 Cry 毒素被广泛应用于农业,以替代化学杀虫剂的使用。我们之前曾报道过一种共表达系统,即在 lac 启动子的控制下,晚期胚胎丰富(LEA)肽增加了 Bt 中杀虫蛋白的表达。使用乳糖诱导 LEA 肽的表达可能是替代异丙基 β-D-硫代半乳糖苷的理想选择,异丙基 β-D-硫代半乳糖苷是最常用于诱导重组蛋白表达的诱导剂。在这项研究中,我们研究了使用乳糖作为诱导剂以获得最佳蛋白表达。我们通过应用一种基于间歇诱导的简单技术观察到了杀虫 Cry 蛋白表达的增强,然后优化了浓度,并将诱导时间从 11 小时调整到 15 小时。我们的数据表明,乳糖的间歇诱导可能是增强细菌蛋白表达的新技术。