Chemically glycosylation improves the stability of an amperometric horseradish peroxidase biosensor

Hernández-Cancel, Griselle; Suazo-Dávila, Dámaris; Medina-Guzmán, Johnsue; Rosado-González, María; Díaz-Vázquez, Liz M.; Griebenow, Kai

doi:10.1016/j.aca.2014.11.008

Cited by 36 publications

(15 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2 b). We observed the presence of two different N atom populations; one at 400 eV attributed to the protonated amine of EDC and the second at 403 eV indicating the presence of the nitrogen of the sulfo-NHS in accordance with previously reported data [ 22 ]. An intense N binding energy peak at 398 eV corresponding to the N 1s 1/2 orbital was observed in the high-resolution spectra of the protein (Fig.…”

Section: Resultssupporting

confidence: 92%

Graphene oxide as a protein matrix: influence on protein biophysical properties

et al. 2015

Self Cite

View full text Add to dashboard Cite

BackgroundThis study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO.ResultsBioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. Enzyme kinetic parameters (km and kcat) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. However, GO produced slight changes in the secondary structure. To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation.ConclusionsIt was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-015-0134-0) contains supplementary material, which is available to authorized users.

show abstract

Section: Resultssupporting

confidence: 92%

Graphene oxide as a protein matrix: influence on protein biophysical properties

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…The nitrogen binding energy is found in the high‐resolution N1s spectrum at 401.8 ± 0.1 eV in Figure S2 (Supporting Information), which can be assigned to the succinimidyl ester intermediate formed by the reaction of EDC/sulfo‐NHS with the carboxylic acid groups of the SAM and is in accordance to previous literature . A second observed nitrogen species at 399.6 ± 0.1 eV can be attributed to the presence of protonated amine of EDC …”

Section: Resultssupporting

confidence: 87%

Characterization of Covalently Bound Anti‐Human Immunoglobulins on Self‐Assembled Monolayer Modified Gold Electrodes

et al. 2017

View full text Add to dashboard Cite

Bioconjugated gold surfaces constitute interesting platforms for biosensing applications. The immobilization of antibodies such as anti-immunoglobulin G and M (anti-IgG and anti-IgM) on gold electrodes via self-assembled monolayers (SAMs) is here studied as a model system for further immunoassays development. The biolayer is characterized by means of X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), a dedicated thin-film transistor (TFT)-based platform and electrochemical surface plasmon resonance (EC-SPR). XPS analysis confirms the presence of all the chemical species involved in the fabrication process as well as the covalent attachment of the antibodies with high reproducibility. Visualization of the biolayer topography by AFM shows nanostructures with a thickness consistent with the actual size of the protein, which is also verified by SPR measurements. EC-SPR allows taking advantage of complementary electrochemical and optical signals during the functionalization steps. Moreover, the functionalization of gold leads to a change in the work function, which is demonstrated in an electrolyte gated thin-film transistor configuration. Such configuration enables also to evaluate the electrostatic changes occurring on the gate that are connected with the threshold voltage shifts. The data support that functional biomodified gold surfaces can be reproducibly prepared, which is a prerequisite for further biosensor development

show abstract

“…The peroxidase stability in extracts derived from potato pulp and brewers' spent grain was higher, compared to other the peroxidase‐containing preparations. Previously, it was reported that the glycosylated HRP used for biosensor construction retained 60% of the initial activity after 100 days of storage at 4°C (Hernández‐Cancel et al, ). HRP immobilized on nano‐Au monolayer retained 60% of the initial activity after 1 month of storage (Lei, Hu, Shen, & Yu, ), and the same enzyme conjugated with starch retained 58 and 24% of the initial activity after 21 and 56 days of storage, respectively (Kagliwal & Singhal, ).…”

Section: Resultsmentioning

confidence: 99%

Study on utilizing solid food industry waste with brewers' spent grain and potato pulp as possible peroxidase sources

et al. 2017

View full text Add to dashboard Cite

This study found that extracts containing ionically bound proteins, isolated from potato pulp and brewers' spent grain, were characterized by high peroxidase activities. The kinetic parameters, namely K m and V max values, were typical for plant peroxidases. Seven peroxidase isoenzymes in potato pulp and two isoenzymes in brewers' spent grain were obtained from their respective ionically bound fractions. Peroxidases from both potato pulp and brewers' spent grain displayed high storage stability, over a 90-day-long storage period, if stored at 2208C with glycerol added to a concentration of 50% or as unsupplemented extracts at 48C. Peroxidase activity was present in the covalently bound fraction of potato pulp, whereas it was absent in the respective fraction of brewers' spent grain. Covalently bound peroxidases from potato pulp displayed high activity, but low stability. Peroxidases extracted from brewers' spent grain and potato pulp, followed the pingpong mechanism and the sequence mechanism, respectively. Practical applicationsPlant peroxidases are widely applicable in various fields of biotechnology and diagnostics. These enzymes are used for biosensor and glucometer construction and in conjugation with antibodies for the purpose of enzyme immunoassays. Using waste products as a peroxidase source provides a cost-friendly alternative to commercially available horseradish peroxidase and gives an opportunity to recycle waste from the food industry-processed plant biomass. Here, the properties of peroxidases from food waste products, namely potato pulp and brewers' spent grain are analyzed. The extracts were characterized by high storage stability and high enzymatic activity, which are two key traits necessary for the practical use of enzyme preparations. K E Y W O R D Sbrewers' spent grain, enzyme storage stability, kinetic studies, peroxidase, potato pulp

show abstract

Chemically glycosylation improves the stability of an amperometric horseradish peroxidase biosensor

Cited by 36 publications

References 43 publications

Graphene oxide as a protein matrix: influence on protein biophysical properties

Graphene oxide as a protein matrix: influence on protein biophysical properties

Characterization of Covalently Bound Anti‐Human Immunoglobulins on Self‐Assembled Monolayer Modified Gold Electrodes

Study on utilizing solid food industry waste with brewers' spent grain and potato pulp as possible peroxidase sources

Contact Info

Product

Resources

About