Part:BBa_K3755003

Mfp5 2*CDS

Two consecutive recombinant mussel foot protein type-5 coding sequences, isolated from Mytilus galloprovincialis, expressed in E.coli BL21.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 457
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

Mussels are magical creatures using their byssus to stick tightly to a variety of substrata that are wet, saline, corroded, and/or fouled by biofilms^[1]. We found that mussel foot protein (Mfp) is the key to the mussel's remarkable adhesion ability. The adhesion of mussel foot proteins is related to several molecular forces, among which the most important are the H-bond, π−π stacking interactions, and metal complexation between the 3,4-dihydroxyphenylalanine(dopa) group in the protein and the hydrophilic group on the material surface^[1,2,7].

Mfp is attractive not just for its great adhesion in water conditions, but for its superior biocompatibility as well^[3]. Up to now, Mfp has been well documented as an excellent wound healing dressing by many articles^[2,4,5]. Our project plans to add Mfp to the hydrogel to increase its mechanical strength and to link broken bone pieces. Since the product needs to be put into the internal environment, which takes high requirements for human safety, Mfp is exactly the best bioglue we can find. By comparison, in the Mfp family, Mfp-5 has the highest dopamine content(∼26 mol %) and the strongest adhesion^[1,6,7]. Therefore, we take the Mfp-5 as the best choice for our adhesion system.

In order to obtain high yield Mfp5 efficiently and conveniently, we chose to express recombinant Mfp5 in E.coli. Luckily, Dong Soo Hwang's team from Pohang University has isolated the Mfp5 sequence from an M. galloprovincialis foot cDNA library and successfully expressed the recombinant Mfp5 in E.coli BL21 in 2003^[8]. Later in 2018, Eugene Kim's team from Washington University in St. Louis referred to Dong Soo Hwang's research, using synthetic DNA with codons optimized for expression and making more amazing designs for Mfp5^[9]. The above results provide us a significant basis for us to produce Mfp5 with high yield and high viscosity.

We learned form Eugene Kim's literatur that repeating the sequence of Mfp5 can lead to higher adhesion^[9]. So we design and registered the 2*Mfp5 CDS into this Part. The complete CDS for 2 repeats of Mfp5 protein expression is registered into the Composite Part part:BBa_K3755012(also named as Mfp5-2) and all the information about protein production and characterization would be shown on that page.

Experiment and Results

We characterized this part in a larger composite part part:BBa_K3755012, please go to that page for more details.

References

[1]Danner EW, Kan Y, Hammer MU, Israelachvili JN, Waite JH. Adhesion of mussel foot protein Mefp-5 to mica: an underwater superglue. Biochemistry. 2012 Aug 21;51(33):6511-8. doi: 10.1021/bi3002538. Epub 2012 Aug 8. PMID: 22873939; PMCID: PMC3428132.
[2]Yang B, Song J, Jiang Y, Li M, Wei J, Qin J, Peng W, Lasaosa FL, He Y, Mao H, Yang J, Gu Z. Injectable Adhesive Self-Healing Multicross-Linked Double-Network Hydrogel Facilitates Full-Thickness Skin Wound Healing. ACS Appl Mater Interfaces. 2020 Dec 30;12(52):57782-57797. doi: 10.1021/acsami.0c18948. Epub 2020 Dec 18. PMID: 33336572.
[3]Huang J, Li H, Wang Q. [Development of double-component rapid curing bioadhesive]. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2018 Dec 25;35(6):921-927. Chinese. doi: 10.7507/1001-5515.201805045. PMID: 30583318.
[4]Liang Y, Zhao X, Hu T, Han Y, Guo B. Mussel-inspired, antibacterial, conductive, antioxidant, injectable composite hydrogel wound dressing to promote the regeneration of infected skin. J Colloid Interface Sci. 2019 Nov 15;556:514-528. doi: 10.1016/j.jcis.2019.08.083. Epub 2019 Aug 24. PMID: 31473541.
[5]Yang Y, Liang Y, Chen J, Duan X, Guo B. Mussel-inspired adhesive antioxidant antibacterial hemostatic composite hydrogel wound dressing via photo-polymerization for infected skin wound healing. Bioact Mater. 2021 Jun 23;8:341-354. doi: 10.1016/j.bioactmat.2021.06.014. PMID: 34541405; PMCID: PMC8427086.
[6]Kord Forooshani P, Lee BP. Recent approaches in designing bioadhesive materials inspired by mussel adhesive protein. J Polym Sci A Polym Chem. 2017 Jan 1;55(1):9-33. doi: 10.1002/pola.28368. Epub 2016 Oct 11. PMID: 27917020; PMCID: PMC5132118.
[7]Lee BP, Messersmith PB, Israelachvili JN, Waite JH. Mussel-Inspired Adhesives and Coatings. Annu Rev Mater Res. 2011 Aug 1;41:99-132. doi: 10.1146/annurev-matsci-062910-100429. PMID: 22058660; PMCID: PMC3207216.
[8]Hwang DS, Yoo HJ, Jun JH, Moon WK, Cha HJ. Expression of functional recombinant mussel adhesive protein Mgfp-5 in Escherichia coli. Appl Environ Microbiol. 2004 Jun;70(6):3352-9. doi: 10.1128/AEM.70.6.3352-3359.2004. PMID: 15184131; PMCID: PMC427802.
[9]Kim E, Dai B, Qiao JB, Li W, Fortner JD, Zhang F. Microbially Synthesized Repeats of Mussel Foot Protein Display Enhanced Underwater Adhesion. ACS Appl Mater Interfaces. 2018 Dec 12;10(49):43003-43012. doi: 10.1021/acsami.8b14890. Epub 2018 Nov 27. PMID: 30480422.