Zafar Muhammad S, Belton David J, Hanby Benjamin, Kaplan David L, Perry Carole C
Interdisciplinary Biomedical Research Centre, School of Science and Technology, Nottingham Trent University , Clifton Lane, Nottingham, NG11 8NS United Kingdom.
Biomacromolecules. 2015 Feb 9;16(2):606-14. doi: 10.1021/bm501667j. Epub 2015 Jan 20.
Bombyx mori (BM) silk fibroin is composed of two different subunits: heavy chain and light chain fibroin linked by a covalent disulfide bond. Current methods of separating the two silk fractions is complicated and produces inadequate quantities of the isolated components for the study of the individual light and heavy chain silks with respect to new materials. We report a simple method of separating silk fractions using formic acid. The formic acid treatment partially releases predominately the light chain fragment (soluble fraction) and then the soluble fraction and insoluble fractions can be converted into new materials. The regenerated original (total) silk fibroin and the separated fractions (soluble vs insoluble) had different molecular weights and showed distinctive pH stabilities against aggregation/precipitation based on particle charging. All silk fractions could be electrospun to give fiber mats with viscosity of the regenerated fractions being the controlling factor for successful electrospinning. The silk fractions could be mixed to give blends with different proportions of the two fractions to modify the diameter and uniformity of the electrospun fibers formed. The soluble fraction containing the light chain was able to modify the viscosity by thinning the insoluble fraction containing heavy chain fragments, perhaps analogous to its role in natural fiber formation where the light chain provides increased mobility and the heavy chain producing shear thickening effects. The simplicity of this new separation method should enable access to these different silk protein fractions and accelerate the identification of methods, modifications, and potential applications of these materials in biomedical and industrial applications.
家蚕(BM)丝素蛋白由两个不同的亚基组成:重链丝素蛋白和轻链丝素蛋白,它们通过共价二硫键相连。目前分离这两种丝组分的方法很复杂,并且所获得的分离组分数量不足以用于研究单个轻链丝和重链丝在新材料方面的特性。我们报道了一种使用甲酸分离丝组分的简单方法。甲酸处理主要部分释放出轻链片段(可溶部分),然后可溶部分和不溶部分都可以转化为新材料。再生的原始(总)丝素蛋白以及分离出的组分(可溶与不可溶)具有不同的分子量,并且基于颗粒带电情况,在抗聚集/沉淀方面表现出独特的pH稳定性。所有丝组分都可以进行静电纺丝以得到纤维毡,再生组分的粘度是成功进行静电纺丝的控制因素。丝组分可以混合以得到两种组分比例不同的共混物,从而改变所形成的静电纺丝纤维的直径和均匀性。含有轻链的可溶部分能够通过使含有重链片段的不溶部分变稀来改变粘度,这可能类似于其在天然纤维形成中的作用,即轻链提供增加的流动性,而重链产生剪切增稠效应。这种新分离方法的简单性应能使人们获取这些不同的丝蛋白组分,并加速对这些材料在生物医学和工业应用中的方法、改性及潜在应用的鉴定。