Department of Biology, Queen's University, Kingston, ON, K7L 3N6, Canada.
Appl Microbiol Biotechnol. 2017 Apr;101(7):2735-2745. doi: 10.1007/s00253-016-8056-9. Epub 2016 Dec 21.
The biosynthesis of quantum dots has been explored as an alternative to traditional physicochemical methods; however, relatively few studies have determined optimal synthesis parameters. Saccharomyces cerevisiae sequentially treated with sodium selenite and cadmium chloride synthesized CdSe quantum dots in the cytoplasm. These nanoparticles displayed a prominent yellow fluorescence, with an emission maximum of approximately 540 nm. The requirement for glutathione in the biosynthetic mechanism was explored by depleting its intracellular content through cellular treatments with 1-chloro-2,4-dinitrobenzene and buthionine sulfoximine. Synthesis was significantly inhibited by both of these reagents when they were applied after selenite treatment prior to the addition of cadmium, thereby indicating that glutathione contributes to the biosynthetic process. Determining the optimum conditions for biosynthesis revealed that quantum dots were produced most efficiently at entry into stationary phase followed by direct addition of 1 mM selenite for only 6 h and then immediately incubating these cells in fresh growth medium containing 3 mM Cd (II). Synthesis of quantum dots reached a maximum at 84 h of reaction time. Biosynthesis of 800-μg g fresh weight cells was achieved. For the first time, significant efforts have been undertaken to optimize each aspect of the CdSe biosynthetic procedure in S. cerevisiae, resulting in a 70% increased production.
量子点的生物合成已被探索作为传统物理化学方法的替代方法;然而,很少有研究确定了最佳的合成参数。酿酒酵母依次用亚硒酸钠和氯化镉处理,在细胞质中合成 CdSe 量子点。这些纳米粒子显示出明显的黄色荧光,发射最大值约为 540nm。通过用 1-氯-2,4-二硝基苯和丁硫氨酸亚砜处理细胞,耗尽其细胞内含量,来探索生物合成机制中谷胱甘肽的需求。在用亚硒酸盐处理后再加入镉之前,这两种试剂都显著抑制了合成,这表明谷胱甘肽有助于生物合成过程。确定生物合成的最佳条件表明,量子点在进入静止期后以最高效率产生,然后直接加入 1mM 亚硒酸盐仅 6 小时,然后立即在含有 3mM Cd(II)的新鲜生长培养基中孵育这些细胞。反应 84 小时时,量子点的合成达到最大值。合成了 800-μg g 鲜重细胞。首次对酿酒酵母中 CdSe 生物合成过程的各个方面进行了优化,产量增加了 70%。