Lin Senzhu, Chen Gefei, Liu Xiangqin, Meng Qing
Institute of Biological Sciences & Biotechnology, Donghua University, Shanghai, 201620, People's Republic of China.
Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada, B3H 4R2.
Biopolymers. 2016 Jul;105(7):385-92. doi: 10.1002/bip.22828.
Hybrid silks hold a great potential as specific biomaterials due to its controlled mechanical properties. To produce fibers with tunable properties, here we firstly made chimeric proteins in vitro, called W2C4CT and W2C8CT, with ligation of MaSp repetitive modules (C) with AcSp modules (W) by intein trans splicing technology from smaller precursors without final yield reduction. Intein mediated chimeric proteins form fibers at a low concentration of 0.4 mg/mL in 50 mM K3 PO4 pH 7.5 just drawn by hand. Hybrid fibers show smoother surface, and also have stronger chemical resistance as compared with fibers from W2CT (W fibers) and mixture of W2CT/C8CT (MHF8 fibers). Fibers from chimeric protein W2C4CT (HFH4) have improved mechanical properties than W fibers; however, with more C modules W2C8CT fibers (HFH8) properties decreased, indicates the length proportion of various modules is very important and should be optimized for fibers with specific properties. Generally, hybrid silks generated via chimeric proteins, which can be simplified by intein trans splicing, has greater potential to produce fibers with tunable properties. Our research shows that intein mediated directional protein ligation is a novel way to make large chimeric spider silk proteins and hybrid silks. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 385-392, 2016.
由于其可控的机械性能,杂合丝作为特定生物材料具有巨大潜力。为了生产具有可调性能的纤维,我们首先在体外制备了嵌合蛋白,称为W2C4CT和W2C8CT,通过内含肽反式剪接技术将MaSp重复模块(C)与AcSp模块(W)从较小的前体连接起来,且最终产量不减。内含肽介导的嵌合蛋白在50 mM K3PO4 pH 7.5的低浓度0.4 mg/mL下就能通过手工拉伸形成纤维。与来自W2CT的纤维(W纤维)和W2CT/C8CT混合物的纤维(MHF8纤维)相比,杂合纤维表面更光滑,化学抗性也更强。来自嵌合蛋白W2C4CT的纤维(HFH4)比W纤维具有更好的机械性能;然而,随着C模块增多,W2C8CT纤维(HFH8)的性能下降,这表明各种模块的长度比例非常重要,应针对具有特定性能的纤维进行优化。一般来说,通过嵌合蛋白产生的杂合丝可通过内含肽反式剪接简化,具有更大潜力生产具有可调性能的纤维。我们的研究表明,内含肽介导的定向蛋白质连接是制备大型嵌合蜘蛛丝蛋白和杂合丝的新方法。© 2016威利期刊公司。生物聚合物105: 385 - 392, 2016。