Expression of Functional Recombinant Mussel Adhesive Protein Mgfp-5 in Escherichia coli

ABSTRACT Mussel adhesive proteins have been suggested as a basis for environmentally friendly adhesives for use in aqueous conditions and in medicine. However, attempts to produce functional and economical recombinant mussel adhesive proteins (mainly foot protein type 1) in several systems have failed. Here, the cDNA coding for Mytilus galloprovincialis foot protein type 5 (Mgfp-5) was isolated for the first time. Using this cDNA, we produced a recombinant Mgfp-5 fused with a hexahistidine affinity ligand, which was expressed in a soluble form in Escherichia coli and was highly purified using affinity chromatography. The adhesive properties of purified recombinant Mgfp-5 were compared with the commercial extracted mussel adhesive Cell-Tak by investigating adhesion force using atomic force microscopy, material surface coating, and quartz crystal microbalance. Even though further macroscale assays are needed, these microscale assays showed that recombinant Mgfp-5 has significant adhesive ability and may be useful as a bioadhesive in medical or underwater environments.

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Characterization and Structure of Mussel Adhesive Proteins

MRS Proceedings ◽

10.1557/proc-174-237 ◽

1989 ◽

Vol 174 ◽

Cited By ~ 1

Author(s):

Richard A. Laursen ◽

Jung-Jung Ou ◽

Xiao-Tong Shen ◽

Michael J. Connors

Keyword(s):

Tandem Repeats ◽

Structural Characteristics ◽

Mytilus Californianus ◽

Adhesive Properties ◽

Marine Animals ◽

Mussel Adhesive Proteins ◽

Sequencing Studies ◽

Mussel Adhesive ◽

Folded Structures ◽

Adhesive Proteins

AbstractMany marine animals such as mussels and barnacles are able to anchor themselves to surfaces through the use of proteinaceous adhesives, which have the unique ability to form strong bonds in a wet environment. With eventual goal of developing wet-surface adhesives, we have carried out gene sequencing studies, using three species of mussel, in order to learn what structural characteristics give these proteins their special properties. Two classes of protein are revealed. Both are characterized by containing about 20% each of lysine and of tyrosine (or DOPA). However, the proteins from Mytilus edulis and Mytilus californianus consist primarily of tandem repeats of variations of the proline-rich decapeptide sequence, AKPSYPPTYK, while the protein from Geukensia demissa is rich in glycine and glutamine and is comprised of 11- and 13-residue variants, e.g., GKPTITYDAGYK, GQQKQTGYDTGYK and GGVQKTGYSAGYK. We propose that the repeat domains have definite folded structures, if not in solution, at least in the condensed (crosslinked) state that gives them their distinctive properties. In this respect, the mussel proteins may resemble collagen, which has both secondary structure and adhesive properties.

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Mass-Production of Practical Mussel Adhesive Protein in Escherichia Coli

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.47-50.857 ◽

2008 ◽

Vol 47-50 ◽

pp. 857-860

Author(s):

Dong Soo Hwang ◽

Seong Hye Lim ◽

Yoon Jeong Yang ◽

Hyung Joon Cha

Keyword(s):

Escherichia Coli ◽

Functional Expression ◽

Pilot Scale ◽

Bioreactor Culture ◽

Batch Bioreactor ◽

Adhesive Ability ◽

Scale Characterization ◽

Mussel Adhesive ◽

Aqueous Conditions ◽

Adhesive Proteins

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 celladhesion analyses, we demonstrate that fp-151 has the potential to be a practical bioadhesive with strong adhesive ability, a simple purification process (~1 g-purified protein per 1 liter-pilot-scale fed-batch bioreactor culture), proper manipulation properties (~330 g/l solubility), and high biocompatibility.

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The adhesive properties of coacervated recombinant hybrid mussel adhesive proteins

Biomaterials ◽

10.1016/j.biomaterials.2010.01.063 ◽

2010 ◽

Vol 31 (13) ◽

pp. 3715-3722 ◽

Cited By ~ 94

Author(s):

Seonghye Lim ◽

Yoo Seong Choi ◽

Dong Gyun Kang ◽

Young Hoon Song ◽

Hyung Joon Cha

Keyword(s):

Adhesive Properties ◽

Mussel Adhesive Proteins ◽

Mussel Adhesive ◽

Adhesive Proteins

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Engineering of Biofilms with a Glycosylation Circuit for Biomaterial Applications

Biomaterials Science ◽

10.1039/d0bm02192j ◽

2021 ◽

Author(s):

Ebru Sahin Kehribar ◽

Musa E İsilak ◽

Eray U. Bozkurt ◽

Jozef Adamcik ◽

Raffaele Mezzenga ◽

...

Keyword(s):

Spider Silk ◽

Post Translational Modification ◽

Adhesive Protein ◽

Mussel Adhesive Proteins ◽

Wide Range ◽

Mussel Adhesive ◽

Adhesive Proteins ◽

Glycosylated Proteins

Glycosylation is a crucial post-translational modification for a wide range of functionalities. Adhesive protein-based biomaterials in nature rely on heavily glycosylated proteins such as spider silk and mussel adhesive proteins....

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Preparation of Sticky Escherichia coli through Surface Display of an Adhesive Catecholamine Moiety

Applied and Environmental Microbiology ◽

10.1128/aem.02223-13 ◽

2013 ◽

Vol 80 (1) ◽

pp. 43-53 ◽

Cited By ~ 18

Author(s):

Joseph P. Park ◽

Min-Jung Choi ◽

Se Hun Kim ◽

Seung Hwan Lee ◽

Haeshin Lee

Keyword(s):

Escherichia Coli ◽

Cell Surface ◽

Communication Systems ◽

Cell Attachment ◽

Surface Display ◽

Content Type ◽

E Coli ◽

Material Surfaces ◽

Mussel Adhesive ◽

Adhesive Proteins

ABSTRACTMussels attach to virtually all types of inorganic and organic surfaces in aqueous environments, and catecholamines composed of 3,4-dihydroxy-l-phenylalanine (DOPA), lysine, and histidine in mussel adhesive proteins play a key role in the robust adhesion. DOPA is an unusual catecholic amino acid, and its side chain is called catechol. In this study, we displayed the adhesive moiety of DOPA-histidine onEscherichia colisurfaces using outer membrane protein W as an anchoring motif for the first time. Localization of catecholamines on the cell surface was confirmed by Western blot and immunofluorescence microscopy. Furthermore, cell-to-cell cohesion (i.e., cellular aggregation) induced by the displayed catecholamine and synthesis of gold nanoparticles on the cell surface support functional display of adhesive catecholamines. The engineeredE. coliexhibited significant adhesion onto various material surfaces, including silica and glass microparticles, gold, titanium, silicon, poly(ethylene terephthalate), poly(urethane), and poly(dimethylsiloxane). The uniqueness of this approach utilizing the engineered stickyE. coliis that no chemistry for cell attachment are necessary, and the ability of spontaneousE. coliattachment allows one to immobilize the cells on challenging material surfaces such as synthetic polymers. Therefore, we envision that mussel-inspired catecholamine yielded stickyE. colithat can be used as a new type of engineered microbe for various emerging fields, such as whole living cell attachment on versatile material surfaces, cell-to-cell communication systems, and many others.

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Complex coacervates based on recombinant mussel adhesive proteins: their characterization and applications

Soft Matter ◽

10.1039/c7sm01735a ◽

2017 ◽

Vol 13 (42) ◽

pp. 7704-7716 ◽

Cited By ~ 20

Author(s):

Hyo Jeong Kim ◽

Byeongseon Yang ◽

Tae Yoon Park ◽

Seonghye Lim ◽

Hyung Joon Cha

Keyword(s):

Practical Applications ◽

Adhesive Protein ◽

Underwater Adhesion ◽

Significant Potential ◽

Mussel Adhesive Proteins ◽

Mussel Adhesive Protein ◽

Mussel Adhesive ◽

Adhesive Proteins

Recombinant mussel adhesive protein-based complex coacervates are an effective underwater adhesive and have significant potential in practical applications that require underwater adhesion.

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