Department of Biomedical Engineering , Tufts University , Medford , Massachusetts 02155 , United States.
Space Biosciences Division, Ames Research Center , National Aeronautics and Space Administration , Moffett Field , California 94035 , United States.
Biomacromolecules. 2018 Sep 10;19(9):3853-3860. doi: 10.1021/acs.biomac.8b00980. Epub 2018 Aug 20.
Despite significant efforts to engineer their heterologous production, recombinant spider silk proteins (spidroins) have yet to replicate the unparalleled combination of high strength and toughness exhibited by natural spider silks, preventing their use in numerous mechanically demanding applications. To overcome this long-standing challenge, we have developed a synthetic biology approach combining standardized DNA part assembly and split intein-mediated ligation to produce recombinant spidroins of previously unobtainable size (556 kDa), containing 192 repeat motifs of the Nephila clavipes dragline spidroin. Fibers spun from our synthetic spidroins are the first to fully replicate the mechanical performance of their natural counterparts by all common metrics, i.e., tensile strength (1.03 ± 0.11 GPa), modulus (13.7 ± 3.0 GPa), extensibility (18 ± 6%), and toughness (114 ± 51 MJ/m). The developed process reveals a path to more dependable production of high-performance silks for mechanically demanding applications while also providing a platform to facilitate production of other high-performance natural materials.
尽管人们已经付出了巨大的努力来对其进行异源生产,但重组蜘蛛丝蛋白(丝氨酸)仍然无法复制天然蜘蛛丝所具有的无与伦比的高强度和韧性组合,这限制了它们在许多机械性能要求高的应用中的使用。为了克服这一长期存在的挑战,我们开发了一种合成生物学方法,结合标准化 DNA 片段组装和分裂内含子介导的连接,以生产以前无法获得的大小(556 kDa)的重组丝氨酸,其中包含 192 个重复的 Nephila clavipes 拖丝丝氨酸重复序列。我们合成的丝氨酸纺制的纤维是首次通过所有常见指标完全复制了天然纤维的机械性能,即拉伸强度(1.03 ± 0.11 GPa)、模量(13.7 ± 3.0 GPa)、伸长率(18 ± 6%)和韧性(114 ± 51 MJ/m)。所开发的工艺为机械性能要求高的应用提供了更可靠的高性能丝的生产途径,同时也为其他高性能天然材料的生产提供了一个平台。