Recombinant production and film properties of full-length hornet silk proteins
Recombinant production and film properties of full-length hornet silk proteins
复制标题
全长黄蜂丝蛋白的重组生产和薄膜特性
DOI：
10.1016/j.actbio.2014.05.013
复制
发表时间：
2014
影响因子：
9.7
通讯作者：
Kambe Y，Sutherland TD，Kameda T
中科院分区：
文献类型：
--
作者：
D. Elmazi;K. Matsuo;T. Oda;M. Ikeda and L. Barolli;Kambe Y，Sutherland TD，Kameda T
在大肠杆菌中制备了vespasimillimahornets蚕茧的四种主要成分Vssilk1-4的全长重组蛋白。在摇瓶中，重组Vssilk蛋白产生160-330 mg重组蛋白1−1。由单个Vssilk蛋白溶液生成的膜具有与天然大黄蜂丝生成的膜相似的二级结构。用单个重组大黄蜂丝蛋白制备的膜与天然大黄蜂丝制备的膜相比具有相似或增强的机械性能，这可能反映了重组蛋白的同质性。测定了各膜zeta （ζ）电位随ph的变化，测定了各膜的等电点pI分别为8.9、9.1、5.0和4.2。4种Vssilk蛋白组合的本地大黄蜂蚕丝的pI值为4.7，与桑蚕蚕丝的pI值相近。生理条件下，由Vssilk1和2制备的膜具有净正电荷，细胞粘附活性显著提高。认为重组大黄蜂丝是一种有价值的细胞培养新材料。
Full-length versions of the four main components of silk cocoons ofVespasimillimahornets, Vssilk1–4, were produced as recombinant proteins inEscherichia coli. In shake flasks, the recombinant Vssilk proteins yielded 160–330 mg recombinant protein l−1. Films generated from solutions of single Vssilk proteins had a secondary structure similar to that of films generated from native hornet silk. The films made from individual recombinant hornet silk proteins had similar or enhanced mechanical performance compared with films generated from native hornet silk, possibly reflecting the homogeneity of the recombinant proteins. The pH-dependent changes in zeta (ζ) potential of each Vssilk film were measured, and isoelectric points (pI) of Vssilk1–4 were determined as 8.9, 9.1, 5.0 and 4.2, respectively. The pI of native hornet silk, a combination of the four Vssilk proteins, was 4.7, a value similar to that ofBombyx morisilkworm silk. Films generated from Vssilk1 and 2 had net positive charge under physiological conditions and showed significantly higher cell adhesion activity. It is proposed that recombinant hornet silk is a valuable new material with potential for cell culture applications.