Expression of functional recombinant mussel adhesive protein type 3A in Escherichia coli
Expression of functional recombinant mussel adhesive protein type 3A in Escherichia coli
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DOI：
10.1021/bp050014e
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发表时间：
2005-05-01
影响因子：
2.9
通讯作者：
Cha, HJ
中科院分区：
文献类型：
--
作者：
Hwang, DS;Gim, Y;Cha, HJ
贻贝黏附蛋白，包括足蛋白3型（fp-3）的20多种变体，已被认为是潜在的环境友好型黏附剂，可用于水环境和医学。本文报道了在大肠杆菌中与六组氨酸亲和配体融合的重组紫贻贝足蛋白3型变体a （Mgfp-3A）的新产物，其亲和层析纯化率接近99%。重组Mgfp-3A与先前报道的重组加洛省分枝杆菌足蛋白5型（Mgfp-5）相比，纯化率更高，在5%乙酸中具有更好的表观溶解度（大规模生产和实际应用的先决条件）。利用石英晶体微天平分析和改进原子力显微镜对纯化后的重组Mgfp-3A与Cell-Tak（一种商业贻贝提取物胶粘剂）和重组Mgfp-5的吸附能力和粘附力进行比较。这些实验表明，重组Mgfp-3A的粘附能力与Cell-Tak相当，但低于重组Mgfp-5。总的来说，这些结果表明重组Mgfp-3A可能作为商业生物粘合剂或医疗或水下环境中的粘合剂成分有用。
Mussel adhesive proteins, including the 20-plus variants of foot protein type 3 (fp-3), have been suggested as potential environmentally friendly adhesives for use in aqueous conditions and in medicine. Here we report the novel production of a recombinant Mytilus galloprovincialis foot protein type 3 variant A (Mgfp-3A) fused with a hexahistidine affinity ligand in Escherichia coli and its similar to 99% purification with affinity chromatography. Recombinant Mgfp-3A showed a superior purification yield and better apparent solubility in 5% acetic acid (prerequisites for large-scale production and practical use) compared to those of the previously reported recombinant M. galloprovincialis foot protein type 5 (Mgfp-5). The adsorption abilities and adhesion forces of purified recombinant Mgfp-3A were compared with those of Cell-Tak (a commercial mussel extract adhesive) and recombinant Mgfp-5 using quartz crystal microbalance analysis and modified atomic force microscopy, respectively. These assays showed that the adhesive ability of recombinant Mgfp-3A was comparable to that of Cell-Tak but lower than that of recombinant Mgfp-5. Collectively, these results indicate that recombinant Mgfp-3A may be useful as a commercial bioadhesive or an adhesive ingredient in medical or underwater environments.