Match!

Self-encapsulation and controlled release of recombinant proteins using novel silica-forming peptides as fusion linkers

Published on Mar 1, 2019in International Journal of Biological Macromolecules4.784
· DOI :10.1016/j.ijbiomac.2018.12.160
Mohamed A.A. Abdelhamid1
Estimated H-index: 1
(Minia University),
Ki Baek Yeo4
Estimated H-index: 4
(KU: Korea University)
+ 1 AuthorsSeung Pil Pack12
Estimated H-index: 12
(KU: Korea University)
Abstract
Abstract Recently, the potential use of biomimetic silica as smart matrices for the auto-encapsulation and controlled release of functional proteins has gained increased interest because of the mild synthesis conditions. Inspired by biological silicification, in this study, we studied novel silica-forming peptides (SFPs), Volp1 and Salp1, to mediate the generation of silica hybrids in vitro. The fusion of SFPs to model fluorescent proteins directed their auto-encapsulation in wet sol-gel silica materials. Furthermore, the SFPs served as affinity linkers for the immobilization of recombinant proteins in silica. Interestingly, the SFP fusion proteins modulated silicic acid polycondensation and allowed for the self-immobilization of SFP fusion proteins in two distinct silica formulations depending on the ionic strength—precipitated silica particles or wet silica gel. The controlled release of Salp1/Volp1 fusion proteins from silica matrices was significantly greater than that of the silaffin R5 fusion proteins. Subsequently, we showed that multiple SFP-tagged proteins homogenously entrapped within a silica matrix could be separately released following pre-incubation with different concentrations of l -arginine solution. These new findings provide a simple and reproducible route for silica hybrid formation for in situ stable auto-encapsulation and the sustained release of recombinant proteins with potential applications in biotechnology.
  • References (47)
  • Citations (3)
📖 Papers frequently viewed together
12 Citations
1 Citations
2005
78% of Scinapse members use related papers. After signing in, all features are FREE.
References47
Newest
#1Marion J. Limo (NTU: Nottingham Trent University)H-Index: 4
#2Anna Sola-Rabada (NTU: Nottingham Trent University)H-Index: 4
Last. Carole C. Perry (NTU: Nottingham Trent University)H-Index: 46
view all 7 authors...
Metallo-oxide (MO)-based bioinorganic nanocomposites promise unique structures, physicochemical properties, and novel biochemical functionalities, and within the past decade, investment in research on materials such as ZnO, TiO2, SiO2, and GeO2 has significantly increased. Besides traditional approaches, the synthesis, shaping, structural patterning, and postprocessing chemical functionalization of the materials surface is inspired by strategies which mimic processes in nature. Would such materi...
6 CitationsSource
#1Damian Pawolski (TUD: Dresden University of Technology)H-Index: 3
#2Christoph Heintze (TUD: Dresden University of Technology)H-Index: 1
Last. Nils Kröger (TUD: Dresden University of Technology)H-Index: 8
view all 5 authors...
Abstract The genetically-controlled formation of complex-shaped inorganic materials by living organisms is an intriguing phenomenon. It illustrates our incomplete understanding of biological morphogenesis and demonstrates the feasibility of ecologically benign routes for materials technology. Amorphous SiO 2 (silica) is taxonomically the most widespread biomineral, with diatoms, a large group of single-celled microalgae, being the most prolific producers. Silica is the main component of diatom c...
5 CitationsSource
#1Mi-Ran Ki (KU: Korea University)H-Index: 7
#2Thi Khoa My Nguyen (KU: Korea University)H-Index: 2
Last. Seung Pil Pack (KU: Korea University)H-Index: 12
view all 4 authors...
Abstract Ferritin (FT), a cage-protein composed of 24 subunits, may carry small molecules inside the core of cage. To enhance the potential of FT as a more functional drug carrier, FT genetically-fused with silaffin R5 peptide at N -terminus (R5FT) and silica-enveloped cage protein (SiO 2 /R5FT) were developed. Doxorubicin (Dox) can be loaded into cage interior by re-assembly of R5FT. The release of Dox loaded in internal cages was retarded by the silica-coated matrix and showed the pH-controlle...
4 CitationsSource
#1Ahmad Seyfaee (University of Newcastle)H-Index: 3
#2Roberto Moreno-Atanasio (University of Newcastle)H-Index: 13
Last. Frances Neville (University of Newcastle)H-Index: 12
view all 3 authors...
Abstract In this work, aqueous solutions of polymeric nuclei based on the polymer polyethyleneimine (PEI), were characterized as a function of pH. The viscosity and zeta potential values of the polymeric solutions were measured and have been related to the degree of protonation of PEI. In addition, the silication of the polymeric nuclei solutions was carried out using trimethoxymethylsilane and the products produced were characterised. The results demonstrate that the zeta potential of the silic...
2 CitationsSource
#1Alexander Kotzsch (TUD: Dresden University of Technology)H-Index: 14
#2Philip Gröger (TUD: Dresden University of Technology)H-Index: 4
Last. Nils Kröger (TUD: Dresden University of Technology)H-Index: 8
view all 7 authors...
Abstract Background Biological mineral formation (biomineralization) proceeds in specialized compartments often bounded by a lipid bilayer membrane. Currently, the role of membranes in biomineralization is hardly understood. Results Investigating biomineralization of SiO 2 (silica) in diatoms we identified Silicanin-1 (Sin1) as a conserved diatom membrane protein present in silica deposition vesicles (SDVs) of Thalassiosira pseudonana . Fluorescence microscopy of GFP-tagged Sin1 enabled, for the...
10 CitationsSource
#1K. G. SprengerH-Index: 5
#2Arushi PrakashH-Index: 3
Last. Jim PfaendtnerH-Index: 24
view all 4 authors...
Biomimetic silica formation, a process that is largely driven by proteins, has garnered considerable interest in recent years due to its role in the development of new biotechnologies. However, much remains unknown of the molecular-scale mechanisms underlying the binding of proteins to biomineral surfaces such as silica, or even of the key residue-level interactions between such proteins and surfaces. In this study, we employ molecular dynamics (MD) simulations to study the binding of R5—a 19-re...
2 CitationsSource
#1Ki Baek Yeo (KU: Korea University)H-Index: 4
#2Mi-Ran Ki (KU: Korea University)H-Index: 7
Last. Seung Pil Pack (KU: Korea University)H-Index: 12
view all 4 authors...
Abstract A synthetic R5 peptide showed the silica precipitation activity when added to silicic acid solution under ambient conditions. However, R5 peptide showed no silica precipitation activity below pH 7, an important pH range for biological applications. Here, we reported new silica-forming peptides (SFP), named EctP1 and EctP2. They showed better silica deposition ability than the R5 peptide. Particularly, at pH 6, EctP1 showed silica deposition activity while R5 did not. EctP1, EctP2, and R...
7 CitationsSource
#1Mohamed Abdel-Hamid (Minia University)H-Index: 39
#2Tsukasa Ikeda (Hiroshima University)H-Index: 30
Last. Akio Kuroda (Hiroshima University)H-Index: 34
view all 7 authors...
We recently reported that the spore coat protein, CotB1 (171 amino acids), from Bacillus cereus mediates silica biomineralization and that the polycationic C-terminal sequence of CotB1 (14 amino acids), designated CotB1p, serves as a silica-binding tag when fused to other proteins. Here, we reduced the length of this silica-binding tag to only seven amino acids (SB7 tag: RQSSRGR) while retaining its affinity for silica. Alanine scanning mutagenesis indicated that the three arginine residues in t...
3 CitationsSource
#1Anne Drathen (Technical University of Dortmund)H-Index: 1
#2Jens Reiber (KIT: Karlsruhe Institute of Technology)H-Index: 2
Last. Susanne Brakmann (Technical University of Dortmund)H-Index: 4
view all 7 authors...
A cell surface-displayed peptide library was employed for the selection of peptides with the capacity to bind fluorescent silica nanoparticles. The outcome of this approach enabled us to identify attributes important for recognition and binding the chosen siliceous material. Two of the peptides that were identified in the course of these experiments were studied in further detail and shown not only to bind silica nanoparticles with high affinity (Kd ∼ 100 nM) but also to induce the enrichment an...
2 CitationsSource
#1Małgorzata Geszke-Moritz (Adam Mickiewicz University in Poznań)H-Index: 9
#2Michał Moritz (PUT: Poznań University of Technology)H-Index: 12
Abstract Four mesoporous siliceous materials such as SBA-16, SBA-15, PHTS and MCF functionalized with (3-aminopropyl)triethoxysilane were successfully prepared and applied as the carriers for poorly water-soluble drug diflunisal. Several techniques including nitrogen sorption analysis, XRD, TEM, FTIR and thermogravimetric analysis were employed to characterize mesoporous matrices. Adsorption isotherms were analyzed using Langmuir, Freundlich, Temkin and Dubinin–Radushkevich models. In order to f...
16 CitationsSource
Cited By3
Newest
#1Mohamed A.A. Abdelhamid (Minia University)H-Index: 1
#2Alaa M.A. Meligy (CNU: Chungnam National University)
Last. Seung Pil Pack (KU: Korea University)H-Index: 12
view all 5 authors...
Abstract Bio-catalysis by enzymes on solid surfaces has been implemented in several practical applications. However, the current methods for efficient enzyme immobilization with retained activity need further development. Herein, a simple, rapid, and economical, bio-affinity-based approach was developed for the direct immobilization with high activity recovery of recombinant Bacillus subtilis catalase (CAT) from the crude lysates. Silaffin-3-derived pentalysine cluster (Sil3K) and its mutant var...
Source
Abstract Elastin-like polypeptides (ELPs), usually as a purification-tag, can be easily expressed and rapidly purified. We found ELPs[V9F-40], which did not contain hydroxyl or charged residues with the pI of 5.52, could biomimetically form silica when they were in ordered structure. The specific activity of ELPs[V9F-40] was 70.89 ± 9.53. Besides, the time needed for the completion of biomimetic silicification was about 138 s, which was only 1/3 of that for other reported peptides. The ELPs@sili...
Source
#1Mi-Ran Ki (KU: Korea University)H-Index: 7
#2Jong Ki Kim (KU: Korea University)
Last. Seung Pil Pack (KU: Korea University)H-Index: 12
view all 6 authors...
Abstract Combinational therapy has been exploited to enhance therapeutic efficiency against cancer by overcoming drug resistance and side-effects or by synergistic effects of multi drugs. However, the conventional co-delivery of combination regimens is often limited by the differences in physicochemical properties among different drugs. Here, to solve this problem, we demonstrated an advanced dual drug delivery system (dDDS) using a R5 peptide-fused ferritin (R5FT). Since the fused R5 can mediat...
Source
#1Ki Ha Min (KU: Korea University)H-Index: 1
#2Ki Baek Yeo (KU: Korea University)H-Index: 4
Last. Seung Pil Pack (KU: Korea University)H-Index: 12
view all 5 authors...
Abstract A novel silica forming peptide (SFP), RSGH was designated on basis of the peptide found from Equus caballus and investigated for its ability to precipitate biosilica under acidic conditions, where biosilica is not easily formed. At pH values of 4, 5 and 6, RSGH showed better performance in the generation of silica particles than the most well-known SFP, silaffin R5, and the amount of the produced biosilica particles increased with the concentration of the peptide. We also demonstrated t...
1 CitationsSource
#1Jong Ki Kim (KU: Korea University)
#2Mohamed A.A. Abdelhamid (Minia University)H-Index: 1
Last. Seung Pil Pack (KU: Korea University)H-Index: 12
view all 3 authors...
Abstract Immobilization of protein, compared to the use of free protein, offers improved stability, easy separation and continuous reusability. However, the classic routes for protein immobilization, based on non-specific adsorption, often negatively affect protein functionality. In this study, EctP1 peptide was explored as a novel short fusion tag for non-covalent adsorption on unmodified solid surfaces, silica and titania. A fusion of EctP1 with bovine carbonic anhydrase (BCA) was employed to ...
Source