Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Biotechnol Bioeng. 2014 Jan;111(1):16-24. doi: 10.1002/bit.25012. Epub 2013 Aug 22.
The toluene-degrading bacterium Acinetobacter sp. Tol 5 shows high adhesiveness mediated by the bacterionanofiber protein AtaA, which is a new member of the trimeric autotransporter adhesin (TAA) family. In contrast to other reported TAAs, AtaA mediates the adhesion of Tol 5 to various abiotic surfaces ranging from hydrophobic plastics to hydrophilic glass and stainless steel. The expression of ataA in industrially relevant bacteria improves their adhesiveness and enables immobilization directly onto support materials. This represents a new method that can be alternated with conventional immobilization via gel entrapment and chemical bonding. In this study, we demonstrate the feasibility of this immobilizing method by utilizing AtaA. As a model case for this method, the indigo producer Acinetobacter sp. ST-550 was transformed with ataA and immobilized on a polyurethane support. The immobilized ST-550 cells were transferred directly to a reaction solution containing indole as the substrate. The immobilized ST-550 cells showed a faster indigo production rate at high concentrations of indole compared with planktonic ST-550 not expressing the ataA gene, implying that immobilization enhanced the tolerance of ST-550 to the substrate indole. As a result, the immobilized ST-550 produced fivefold higher levels of indigo than planktonic ST-550. These results proved that AtaA is useful for bacterial immobilization.
甲苯降解菌不动杆菌 sp. Tol 5 表现出高粘附性,这是由菌毛纤维蛋白 AtaA 介导的,AtaA 是三聚体自转运体粘附素(TAA)家族的新成员。与其他报道的 TAAs 不同,AtaA 介导 Tol 5 对各种非生物表面的粘附,范围从疏水性塑料到亲水性玻璃和不锈钢。ataA 在工业相关细菌中的表达提高了它们的粘附性,并能够直接固定在支持材料上。这代表了一种可以替代传统凝胶包埋和化学结合的固定化方法。在这项研究中,我们通过利用 AtaA 证明了这种固定化方法的可行性。作为该方法的模型案例,将吲哚生产菌不动杆菌 sp. ST-550 转化为 ataA 并固定在聚氨酯载体上。固定化的 ST-550 细胞直接转移到含有吲哚作为底物的反应溶液中。与不表达 ataA 基因的浮游 ST-550 相比,固定化的 ST-550 细胞在吲哚浓度较高时表现出更快的靛蓝生产速率,这意味着固定化增强了 ST-550 对底物吲哚的耐受性。结果,固定化的 ST-550 产生的靛蓝比浮游 ST-550 高出五倍。这些结果证明 AtaA 可用于细菌固定化。