Efficient Immobilization of Protein on Monodispersed Colloidal Silica Particles Modified by Copolymers of Maleic Anhydride and Styrene or Methyl Methacrylate

Yoshinaga, Kohji; Kondo, Kimihiko; Kondo, Akihiko

doi:10.1295/polymj.27.98

Cited by 11 publications

(4 citation statements)

References 8 publications

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“…One kind of successful sample is the commercial glucose electrodes, which have been developed based on immobilized glucose oxidase . So far, research of the immobilization of enzymes on different matrixes, such as polymers, sol−gels, phospholipids, mesoporous molecular sieves, and self-assembled monolayers, have attracted more and more attention and achieved many significant results. These hosts provide a protective environment for enzymes and improve the stability of immobilized enzymes, although the structures of most selected support matrixes and the formed composites are not well-ordered.…”

Section: Introductionmentioning

confidence: 99%

Layered Structural Heme Protein Magadiite Nanocomposites with High Enzyme-like Peroxidase Activity

et al. 2004

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Layered nanocomposites of global myoglobin (Mb)-magadiite and hemoglobin (Hb)magadiite with the interlayer distances of 43.2 and 68.2 Å, respectively, have been successfully prepared by using tetrabutylammonium (TBA)-magadiite as the intermediate under mild conditions. The structures, details of the interaction between the host magadiite and guest proteins, and saturated bounding amounts of Mb and Hb were characterized and studied by many techniques. The enzyme activity study showed that the fresh intercalated Mb and Hb nanocomposites retained most enzyme-like peroxidase activity and about 25.0% of activities based on the formation amounts of tetramers were kept after 30 min of reaction when these nanocomposites were stored for 1 month at room temperature. The relations between the structures and the peroxidase activities were discussed based on Michaelis-Menten model analyses. Such stable and high enzyme-like peroxidase activity for immobilized Hb and Mb provide potential application in biosensors and enzyme reactors.

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Section: Introductionmentioning

confidence: 99%

Layered Structural Heme Protein Magadiite Nanocomposites with High Enzyme-like Peroxidase Activity

et al. 2004

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“…Some examples have included polymers, sol-gels, surfactants and hydrogels. [4][5][6][7][8] Recently, inorganic nanoparticles have also been used to immobilize proteins or enzymes and demonstrated a unique and suitable microenvironment for the proteins. 9 Studies of direct electrochemistry of protein or enzymes at electrodes can serve as a basis for building electrochemical biosensors, enzymatic bioreactors, and biomedical devices.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical biosensors utilising electron transfer in heme proteins immobilised on Fe₃O₄nanoparticles

Cao

2003

Analyst

147

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Fe3O4 nanoparticles cast on pyrolytic graphite (PG) electrodes were used to immobilize hemoglobin (Hb), myoglobin (Mb) and horseradish peroxidase (HRP). The Fe3O4 nanoparticles provided a favorable microenvironment for the proteins to directly transfer electrons with electrodes. The protein-Fe3O4 films were used to electrochemically catalyze the reduction of oxygen, trichloroacetic acid, nitrite and hydrogen peroxide, and showed a potential applicability in fabricating biosensors. Transmission electron microscopy (TEM), UV-visible absorption and reflectance absorption infrared (RAIR) spectroscopy, and cyclic and square wave voltammetry, were used to characterize the films.

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“…The reasonably fast time-scales for the synthesis of the enzymelipid composites and the enzyme-friendly encapsulation conditions are a major improvement over other techniques currently being investigated. 1,3,4,6 Two of us (A. G. and C. D.) thank the Council for Scientific and Industrial Research (CSIR), Government of India, for financial assistance. Fig.…”

confidence: 99%

“…; retention of the native protein structure and accessibility of the encapsulated proteins to cofactors, substrates and redox agents is a problem of current interest especially where application in biosensors/ biocatalysis is sought. Proteins have been immobilized in phospholipid bilayers, 1 on self-assembled monolayers (SAMs), 2 in silicate sol-gels, 3 in polymer matrices, 4 in Langmuir-Blodgett films, 5 within the galleries of a-zirconium phosphates 6 as well as polymer microspheres. 7 Developing on our earlier work on the spontaneous self-organization of fatty acid salts 8 and electrostatic assembly of colloidal nanoparticles, 9,10 we show here, that the proteolytic enzyme pepsin (which occurs in the gastric juice of all mammals), can be encapsulated via electrostatic interaction in thermally evaporated fatty lipid matrices (octadecylamine, ODA) by simple immersion of the lipid film in the protein solution under extremely mild preparation conditions (Schematic 1, inset of Fig.…”

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confidence: 99%

Encapsulation and biocatalytic activity of the enzyme pepsin in fatty lipid films by selective electrostatic interactions

et al. 2000

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The encapsulation of pepsin by electrostatically controlled diffusion from solution into thermally evaporated fatty amine films is described and the catalytic activity of the immobilized enzyme on hemoglobin is investigated.The entrapment of proteins in different inert matrices with the aim of protecting the proteins against microbial degradation, hydrolysis, autoproteolysis, deamidation, etc.; retention of the native protein structure and accessibility of the encapsulated proteins to cofactors, substrates and redox agents is a problem of current interest especially where application in biosensors/ biocatalysis is sought. Proteins have been immobilized in phospholipid bilayers, 1 on self-assembled monolayers (SAMs), 2 in silicate sol-gels, 3 in polymer matrices, 4 in Langmuir-Blodgett films, 5 within the galleries of a-zirconium phosphates 6 as well as polymer microspheres. 7 Developing on our earlier work on the spontaneous self-organization of fatty acid salts 8 and electrostatic assembly of colloidal nanoparticles, 9,10 we show here, that the proteolytic enzyme pepsin (which occurs in the gastric juice of all mammals), can be encapsulated via electrostatic interaction in thermally evaporated fatty lipid matrices (octadecylamine, ODA) by simple immersion of the lipid film in the protein solution under extremely mild preparation conditions (Schematic 1, inset of Fig. 1). The encapsulated enzyme showed good biocatalytic activity using hemoglobin as the substrate. The biocatalytic activity was determined by estimating the amount of acidsoluble tyrosine and tryptophan residues released by reaction of the encapsulated pepsin on hemoglobin.Pepsin (molecular weight = 37400; pI ≈ 1 where pI represents the isoelectric point) 11 was obtained from Sigma Chemicals and used as received. A 10 26 M solution of pepsin was prepared in glycine-HCl buffer (0.05 M, pH 3), close to the pH value at which pepsin exhibits maximum catalytic activity. 250 and 1000 Å thick ODA (Aldrich) films were deposited by thermal evaporation on gold coated AT cut quartz crystals (for quartz crystal microgravimetry (QCM) measurements) and Si (111) substrates (for FTIR and activity measurements) in an Edwards E306A chamber. Fig. 1 shows a plot of the QCM mass uptake recorded ex situ from a 250 Å thick ODA film as a function of time of immersion in the pepsin solution. Details of the QCM measurement procedure may be obtained from our earlier reports. 9,10 It can be seen that the diffusion of pepsin into the lipid matrix is extremely rapid with maximum protein uptake being accomplished within 10 min of immersion. The small time scale for encapsulation of the enzyme in the lipid matrix is a particularly attractive feature of this approach and considerably improves upon the days to weeks timeframe required in other techniques for synthesis of such biocomposites. 3,6 At pH 3, the ODA matrix is positively charged (pK B of ODA = 10.5) while the pepsin molecules are negatively charged (pI of pepsin ≈ 1.0 ) thereby leading to attractive electrostatic interacti...

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Efficient Immobilization of Protein on Monodispersed Colloidal Silica Particles Modified by Copolymers of Maleic Anhydride and Styrene or Methyl Methacrylate

Cited by 11 publications

References 8 publications

Layered Structural Heme Protein Magadiite Nanocomposites with High Enzyme-like Peroxidase Activity

Layered Structural Heme Protein Magadiite Nanocomposites with High Enzyme-like Peroxidase Activity

Electrochemical biosensors utilising electron transfer in heme proteins immobilised on Fe₃O₄nanoparticles

Encapsulation and biocatalytic activity of the enzyme pepsin in fatty lipid films by selective electrostatic interactions

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Efficient Immobilization of Protein on Monodispersed Colloidal Silica Particles Modified by Copolymers of Maleic Anhydride and Styrene or Methyl Methacrylate

Cited by 11 publications

References 8 publications

Layered Structural Heme Protein Magadiite Nanocomposites with High Enzyme-like Peroxidase Activity

Layered Structural Heme Protein Magadiite Nanocomposites with High Enzyme-like Peroxidase Activity

Electrochemical biosensors utilising electron transfer in heme proteins immobilised on Fe3O4nanoparticles

Encapsulation and biocatalytic activity of the enzyme pepsin in fatty lipid films by selective electrostatic interactions

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Electrochemical biosensors utilising electron transfer in heme proteins immobilised on Fe₃O₄nanoparticles