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Journal of Chemical Sciences

, 130:145 | Cite as

Stimuli-responsive, protein hydrogels for potential applications in enzymology and drug delivery\(^{\S }\)

  • Ajith Pattammattel
  • Bobbi S Stromer
  • Clive Baveghems
  • Kyle Benson
  • Challa V Kumar
Regular Article
  • 34 Downloads

Abstract

Enhancing the stability of enzymes for sensing or biocatalysis applications is still an unmet challenge. Ordinary paper is a very attractive support for anchoring enzymes but enzyme attachment to cellulose without surface activation is still another challenge. To make progress toward these goals, we developed a simple method to prepare highly active and stable enzyme-hydrogels within the mesh of the cellulose fibers of paper. A mixture of the desired enzyme, bovine serum albumin (BSA) and arginine were reacted with carbodiimide to form stable hydrogels. A set of critical concentrations \((\hbox {BSA}([\hbox {BSA}]_{0}) \,{\ge }1~\hbox {mM})\), \([\hbox {carbodiimide}]_{0} \ge 100\) mM and [amino acid] \(\ge 100\) mM) were required to form transparent hydrogels. The thermal reversibility of gelation proved that the gels are stabilized by non-covalent bonding interactions between the BSA oligomers that were formed via covalent interactions. Both dynamic light scattering and SDS-PAGE studies, under pre-gelation conditions, support idea that one BSA oligomeric unit contained 40–70 protein molecules. Scanning electron micrographs, thermogravimetry and swelling studies suggest that the formation of water cavities inside the cross-linked gel matrix, where the water mass was 7–8 times higher than that of the protein and the free amino acid used as a linker/spacer. Due to the higher water content and benign gelation conditions, active enzymes could be incorporated into the gel structure during the synthesis. Hydrogels, thus, embedded with glucose oxidase (GOx) and horseradish peroxide (HRP) showed catalytic activity towards glucose, where efficient channeling of hydrogen peroxide from GOx to HRP was observed (70% efficiency in initial rate compared to free enzymes in solution). Moreover, the enzymes retained their activity after pasting the hydrogel onto ordinary paper, which was demonstrated as a glucose sensing platform with a detection limit of 5 mM glucose. Trypsin embedded in the gel showed temperature dependent self-degradation by utilizing optimum protease activity at \(37\,{^{\circ }}\hbox {C}\). The temperature-triggered degradation of the gel can be used as a drug delivery vehicle, which was demonstrated using a reporter dye. The hydrogel made of a completely proteinaceous material that releases drugs at body temperature but bound to the matrix at room temperature (\(25\,{^{\circ }}\hbox {C}\)) is useful for noninvasive drug delivery platforms. The biocompatibility and non-thermal synthetic route for the hydrogel makes it a superior material for incorporation of temperature sensitive enzymes, drug molecules or nucleic acids, for a diverse set of applications.

Graphical Abstract Synopsis

Synthesis, characterization and applications of multifunctional, biologically benign, protein-derived hydrogels are reported here. Protein hydrogels are formed by both covalent and noncovalent interactions between amino acid residues of the constituents. Active enzymes embedded inside the gel matrix were utilized for biosensing and drug delivery applications.

Keywords

Bovine serum albumin glucose oxidase dynamic light scattering SDS PAGE rhodamine B glucose detection paper guaiacol 

Notes

Acknowledgements

We thank the National Science Foundation (EAGER, DMR-1401879) and the University of Connecticut OVPR Research Excellence Program for financial support of this work. CB is currently a postdoctoral fellow at Boston University Medical School funded by the Multidisciplinary Training in Cardiovascular Research grant, NIH T32 HL07224.

Supplementary material

12039_2018_1538_MOESM1_ESM.pdf (149 kb)
Supplementary material 1 (pdf 149 KB)

References

  1. 1.
    Caló E and Khutoryanskiy V V 2015 Biomedical applications of hydrogels: a review of patents and commercial products Eur. Polym. J. 65 252CrossRefGoogle Scholar
  2. 2.
    Chai Q, Jiao Y and Yu X 2017 Hydrogels for biomedical applications: their characteristics and the mechanisms behind them Gels 3 6CrossRefGoogle Scholar
  3. 3.
    Owens G J, Singh R K, Foroutan F, Alqaysi M, Han C M, Mahapatra C, Kim H W and Knowles JC 2016 Sol–gel based materials for biomedical applications Progress Mater. Sci. 77 1CrossRefGoogle Scholar
  4. 4.
    Langer R and Tirrell D A 2004 Designing materials for biology and medicine Nature 428 487CrossRefGoogle Scholar
  5. 5.
    Van Vlierberghe S, Dubruel P and Schacht E 2011 Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review Biomacromolecules 12 1387CrossRefGoogle Scholar
  6. 6.
    Ahmed E M 2015 Hydrogel: preparation, characterization, and applications: a review J. Adv. Res6 105CrossRefGoogle Scholar
  7. 7.
    Gagner J E, Kim W and Chaikof E L 2014 Designing protein-based biomaterials for medical applications Acta Biomater10 1542CrossRefGoogle Scholar
  8. 8.
    Lee K Y and David J M 2001 Hydrogels for tissue engineering Chem. Rev.  101 1869CrossRefGoogle Scholar
  9. 9.
    Payne J W 1973 Polymerization of proteins with glutaraldehyde. Soluble molecular-weight markers Biochem. J.  135 867CrossRefGoogle Scholar
  10. 10.
    Nakajima N and Ikada Y 1995 Mechanism of amide formation by carbodiimide for bioconjugation in aqueous media Bioconj. Chem. 6 123CrossRefGoogle Scholar
  11. 11.
    Benson K, Ghimire A, Pattammattel A and Kumar C V 2017 Protein biophosphors: biodegradable, multifunctional, protein-based hydrogel for white emission, sensing, and pH detection Adv. Funct. Mater. 27 1702955CrossRefGoogle Scholar
  12. 12.
    Ghimire A, Pattammattel A, Maher C E, Kasi R M and Kumar C V 2017 Three-dimensional, enzyme biohydrogel electrode for improved bioelectrocatalysis. ACS Appl. Mater. Interfaces 9 42556CrossRefGoogle Scholar
  13. 13.
    Raybould A, Burns A and Hamer M 2014 high concentrations of protein test substances may have non-toxic effects on daphnia magna: implications for regulatory study designs and ecological risk assessments for gm crops GM Crops Food 5 296CrossRefGoogle Scholar
  14. 14.
    Nunn B L, Norbeck A and Keil R G 2003 Hydrolysis patterns and the production of pepetide intermediates during protein degreadation in marine systems Marine Chem. 83 59CrossRefGoogle Scholar
  15. 15.
    Lill J R, Ingles E S, Liu P S, Pham V and Sandoval W N 2007 Microwave assisted proteomics Mass Spectrom. Rev. 26 657CrossRefGoogle Scholar
  16. 16.
    Lohcharoenkal W, Wang L, Chen Y C and Rojanasakul Y 2014 Protein nanoparticles as drug delivery carriers for cancer therapy Biomed. Res. Int. 2014 180549CrossRefGoogle Scholar
  17. 17.
    Xing Q, Yates K, Vogt C, Qian Z, Frost M C and Zhao F 2014 Increasing mechanical strength of gelatin hydrogels by divalent metal ion removal Sci. Rep. 4 4706CrossRefGoogle Scholar
  18. 18.
    Novak M J, Pattammattel A, Koshmerl B, Puglia M, Williams C and Kumar C V 2016 “Stable-on-the-Table” Enzymes: Engineering the Enzyme–Graphene Oxide Interface for Unprecedented Kinetic Stability of the Biocatalyst ACS Catal. 6 339CrossRefGoogle Scholar
  19. 19.
    Ma X, Sun X, Hargrove D, Chen J, Song D, Dong Q, Lu X, Fan T H, Fu Y and Lei Y 2016 A biocompatible and biodegradable protein hydrogel with green and red autofluorescence: preparation, characterization and in vivo biodegradation tracking and modeling Sci. Rep. 6 19370CrossRefGoogle Scholar
  20. 20.
    Gebregeorgis A, Bhan C, Wilson O and Raghavan D 2013 Characterization of silver/bovine serum albumin (Ag/BSA) nanoparticles structure: morphological, compositional, and interaction studies J. Colloid Interface Sci.  389 31CrossRefGoogle Scholar
  21. 21.
    Shi Q, Zhou Y and Sun Y 2005 Influence of pH and ionic strength on the steric mass-action model parameters around the isoelectric point of protein Biotechnol. Prog. 21 516CrossRefGoogle Scholar
  22. 22.
    Yasuda K, Nakamura R and Hayakawa S 1986 Factors affecting heat-induced gel formation of bovine serum albumin J. Food Sci. 51 1289CrossRefGoogle Scholar
  23. 23.
    Majorek K A, Porebski P J, Dayal A, Zimmerman M D, Jablonska K, Stewart A J, Chruszcz M and Minor W 2012 Structural and immunologic characterization of bovine, horse, and rabbit serum albumins Mol. Immunol. 52 174CrossRefGoogle Scholar
  24. 24.
    Flecha F L G and Levi V 2003 Determination of the molecular size of BSA by fluorescence anisotropy Biochem. Mol. Biol. Edu.  31 319CrossRefGoogle Scholar
  25. 25.
    Riccardi C M, Mistri D, Hart O, Anuganti M, Lin Y, Kasi R M and Kumar C V 2016 Covalent interlocking of glucose oxidase and peroxidase in the voids of paper: enzyme–polymer “spider webs” Chem. Commun.  52 2593CrossRefGoogle Scholar
  26. 26.
    Li S, Su Y, Luo W and Hong M 2010 Water-protein interactions of an arginine-rich membrane peptide in lipid bilayers investigated by solid-state nuclear magnetic resonance spectroscopy J. Phys. Chem. B 114 4063CrossRefGoogle Scholar
  27. 27.
    Knipe J M, Chen F and Peppas N A 2015 Enzymatic biodegradation of hydrogels for protein delivery targeted to the small intestine Biomacromolecules 16 962CrossRefGoogle Scholar
  28. 28.
    Olsen J V, Ong S E and Mann M 2004 Trypsin cleaves exclusively C-terminal to arginine and lysine residues Mol. Cell Proteomics 3 608CrossRefGoogle Scholar

Copyright information

© Indian Academy of Sciences 2018

Authors and Affiliations

  • Ajith Pattammattel
    • 1
  • Bobbi S Stromer
    • 1
  • Clive Baveghems
    • 1
  • Kyle Benson
    • 1
  • Challa V Kumar
    • 1
  1. 1.Department of Chemistry and Molecular Cell Biology, Institute of Material SciencesUniversity of ConnecticutStorrsUSA

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