Biosystems Engineering Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres Bello, República 239, 8370146 Santiago, Chile.
BioNanotechnology and Microbiology Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres Bello, República 239, 8370146 Santiago, Chile.
J Biotechnol. 2017 Dec 20;264:29-37. doi: 10.1016/j.jbiotec.2017.10.013. Epub 2017 Oct 19.
Microbial polymers and nanomaterials production is a promising alternative for sustainable bioeconomics. To this end, we used Pseudomonas putida KT2440 as a cell factory in batch cultures to coproduce two important nanotechnology materials- medium-chain-length (MCL)-polyhydroxyalkanoates (PHAs) and CdS fluorescent nanoparticles (i.e. quantum dots [QDots]). Due to high cadmium resistance, biomass and PHA yields were almost unaffected by coproduction conditions. Fluorescent nanocrystal biosynthesis was possible only in presence of cysteine. Furthermore, this process took place exclusively in the cell, displaying the classical emission spectra of CdS QDots under UV-light exposure. Cell fluorescence, zeta potential values, and particles size of QDots depended on cadmium concentration and exposure time. Using standard PHA-extraction procedures, the biosynthesized QDots remained associated with the biomass, and the resulting PHAs presented no traces of CdS QDots. Transmission electron microscopy located the synthesized PHAs in the cell cytoplasm, whereas CdS nanocrystals were most likely located within the periplasmic space, exhibiting no apparent interaction. This is the first report presenting the microbial coproduction of MCL-PHAs and CdS QDots in P. putida KT2440, thus constituting a foundation for further bioprocess developments and strain engineering towards the efficient synthesis of these highly relevant bioproducts for nanotechnology.
微生物聚合物和纳米材料的生产是可持续生物经济的一种有前途的替代方案。为此,我们使用恶臭假单胞菌 KT2440 作为细胞工厂在分批培养中共同生产两种重要的纳米技术材料——中链长度(MCL)-聚羟基脂肪酸酯(PHA)和 CdS 荧光纳米粒子(即量子点 [QDs])。由于高镉抗性,生物量和 PHA 产量几乎不受共生产条件的影响。只有在半胱氨酸存在的情况下才能进行荧光纳米晶体的生物合成。此外,该过程仅在细胞内进行,在暴露于紫外线下时显示出 CdS QDots 的典型发射光谱。细胞荧光、Zeta 电位值和 QDots 的粒径取决于镉浓度和暴露时间。使用标准 PHA 提取程序,生物合成的 QDots 仍然与生物质相关联,并且得到的 PHAs 没有 CdS QDots 的痕迹。透射电子显微镜将合成的 PHAs 定位在细胞质中,而 CdS 纳米晶体很可能位于周质空间内,没有明显的相互作用。这是首次报道恶臭假单胞菌 KT2440 中共同生产 MCL-PHAs 和 CdS QDots,为进一步的生物工艺开发和菌株工程奠定了基础,以高效合成这些对纳米技术具有重要意义的生物产品。