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实用的重组杂交贻贝生物粘合剂fp-151

Practical recombinant hybrid mussel bioadhesive fp-151.

作者信息

Hwang Dong Soo, Gim Youngsoo, Yoo Hyo Jin, Cha Hyung Joon

机构信息

Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea.

出版信息

Biomaterials. 2007 Aug;28(24):3560-8. doi: 10.1016/j.biomaterials.2007.04.039. Epub 2007 May 3.

Abstract

Mussel adhesive proteins (MAPs) have received increased attention as potential environmentally friendly adhesives under aqueous conditions and in medicine. However, attempts to produce functional recombinant MAPs (mainly foot protein type 1, fp-1) by several expression systems have failed. Even though we previously reported a functional expression of recombinant foot protein type 5 (fp-5) with significant adhesive ability in Escherichia coli, its practical use was limited by several problems such as low production yield, low purification yield, and high levels of post-purification insolubility. Here, to overcome these limitations, we designed and constructed the novel type of hybrid mussel bioadhesive fp-151, a fusion protein comprising six fp-1 decapeptide repeats at each fp-5 terminus. Using micro- and bulk-scale characterization and mammalian cell-adhesion analyses, we demonstrate that fp-151 has the potential to be a practical bioadhesive with strong adhesive ability, a simple purification process ( approximately 1g-purified protein per 1l-pilot-scale fed-batch bioreactor culture), proper manipulation properties ( approximately 330g/l solubility), and high biocompatibility.

摘要

贻贝粘附蛋白(MAPs)作为在水性条件下及医学领域中潜在的环境友好型粘合剂,已受到越来越多的关注。然而,利用多种表达系统生产功能性重组MAPs(主要是足蛋白1型,fp - 1)的尝试均告失败。尽管我们之前报道了重组足蛋白5型(fp - 5)在大肠杆菌中具有显著粘附能力的功能性表达,但其实际应用受到诸如低产量、低纯化率以及纯化后高不溶性等几个问题的限制。在此,为克服这些限制,我们设计并构建了新型的杂合贻贝生物粘合剂fp - 151,这是一种在每个fp - 5末端包含六个fp - 1十肽重复序列的融合蛋白。通过微观和宏观尺度表征以及哺乳动物细胞粘附分析,我们证明fp - 151有潜力成为一种实用的生物粘合剂,具有强大的粘附能力、简单的纯化过程(每1升中试规模补料分批生物反应器培养可获得约1克纯化蛋白)、合适的操作特性(约330克/升的溶解度)以及高生物相容性。

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