Bulk adhesive strength of recombinant hybrid mussel adhesive protein
Bulk adhesive strength of recombinant hybrid mussel adhesive protein
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DOI：
10.1080/08927010802563108
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发表时间：
2009-01-01
期刊：
影响因子：
2.7
通讯作者：
Wilker, Jonathan J.
中科院分区：
文献类型：
--
作者：
Cha, Hyung Joon;Hwang, Dong Soo;Wilker, Jonathan J.
贻贝粘附蛋白（MAPs）作为一种潜在的生物医学和环境友好型粘合剂，受到越来越多的关注。然而，不经济的提取和不成功的遗传生产严重限制了MAP的实际应用。开发新的粘合剂需要获得大量的材料和批量机械性能的演示。在此之前，作者设计了fp-151，一种融合蛋白，其在每个MAP 5型（fp-5）末端包含6个MAP 1型（fp-1）十肽重复序列，并在大肠杆菌中成功表达。该重组杂合蛋白具有高效表达、纯化简单、生物相容性好、微尺度粘附能力强等特点。在本工作中，在空气中进行的半纯化（90%纯度）的融合fp-151的本体粘合性能的调查。表达的未修饰的重组fp-151含有酪氨酸残基，并显示出显著的剪切粘附力（0.33MPa）。在酶促氧化酪氨酸残基为1 - 3，4-二羟基苯丙氨酸（DOPA）基团后，粘附强度增加（0.45MPa）。添加交联剂如铁（III）、锰（III）和高碘酸盐（IO 4-）通常增强粘附性，尽管添加太多会降低粘附性。在所考察的三种交联剂中，非金属氧化剂高碘酸盐的剪切粘附力最高（0.86MPa）。此外，发现可以通过向样品添加重量来增加粘合强度。发现的最高粘合强度是与高碘酸盐交联的含DOPA的fp-151的粘合强度，并且具有施加到样品上的重量（1.06MPa）。两者合计，第一个批量规模的粘附力测量的表达重组杂交贻贝粘附蛋白。
Mussel adhesive proteins (MAPs) have received increased attention as potential biomedical and environmental friendly adhesives. However, practical application of MAPs has been severely limited by uneconomical extraction and unsuccessful genetic production. Developing new adhesives requires access to large quantities of material and demonstrations of bulk mechanical properties. Previously, the authors designed fp-151, a fusion protein comprised of six MAP type 1 (fp-1) decapeptide repeats at each MAP type 5 (fp-5) terminus and successfully expressed it in Escherichia coli. This recombinant hybrid protein exhibited high-level expression, a simple purification and high biocompatibility as well as strong adhesive ability on a micro-scale. In the present work, investigations on the bulk adhesive properties of semi-purified (90% purity) fusion fp-151 were performed in air. The unmodified recombinant fp-151, as expressed, contains tyrosine residues and showed significant shear-adhesive forces (0.33MPa). Adhesion strength increased (0.45MPa) after enzymatic oxidation of tyrosine residues to l-3,4-dihydroxyphenylalanine (DOPA) groups. Addition of cross-linkers such as iron(III), manganese(III) and periodate (IO4-) generally enhanced adhesion, although too much addition decreased adhesion. Among the three cross-linking reagents examined, the non-metallic oxidant periodate showed the highest shear-adhesive forces (0.86MPa). In addition, it was found that adhesive strengths could be increased by adding weights to the samples. The highest adhesion strength found was that of DOPA-containing fp-151 cross-linked with periodate and having weights applied to the samples (1.06MPa). Taken together, the first bulk-scale adhesive force measurements are presented for an expressed recombinant hybrid mussel adhesive protein.