In situ structure and activity studies of an enzyme adsorbed on spectroscopically undetectable particles

S. Koutsopoulos, A.M. Tjeerdsma, J.F.T. van Lieshout, J. van der Oost, W. Norde

Research output: Contribution to journalArticleAcademicpeer-review

19 Citations (Scopus)

Abstract

The structural characteristics and the activity of a hyperthermophilic endoglucanase were investigated upon adsorption. Silica (hydrophilic) and Teflon (hydrophobic) surfaces were selected for the study. The materials were specially designed so that the interaction of the particles with light was negligible, and the enzyme conformation in the adsorbed state was monitored in situ. The adsorption isotherms were determined, and the adsorbed endoglucanase was studied using a number of spectroscopic techniques, enzymatic activity tests, and dynamic light scattering. Experiments were performed at pH values below, at, and above the isoelectric point of the enzyme. It was shown that the enzyme adsorbed on the hydrophobic surface of Teflon with higher affinity as compared to the hydrophilic silica nanoparticles. In all cases, adsorption was followed by (slight) changes in the secondary structure resulting in decreased -structural content. The changes were more profound upon adsorption on Teflon. The adsorbed enzyme remained active in the adsorbed state in spite of the structural changes induced when interacting with the surfaces
Original languageEnglish
Pages (from-to)1176-1184
JournalBiomacromolecules
Volume6
Issue number3
DOIs
Publication statusPublished - 2005

Fingerprint

Polytetrafluoroethylene
Enzymes
Polytetrafluoroethylenes
Cellulase
Adsorption
Silicon Dioxide
Silica
Dynamic light scattering
Adsorption isotherms
Conformations
Nanoparticles
Experiments

Keywords

  • archaeon pyrococcus-furiosus
  • solid-liquid interfaces
  • circular-dichroism
  • proteins
  • adsorption
  • endo-beta-1,3-glucanase
  • extremophiles
  • surfaces
  • features
  • sugar

Cite this

Koutsopoulos, S. ; Tjeerdsma, A.M. ; van Lieshout, J.F.T. ; van der Oost, J. ; Norde, W. / In situ structure and activity studies of an enzyme adsorbed on spectroscopically undetectable particles. In: Biomacromolecules. 2005 ; Vol. 6, No. 3. pp. 1176-1184.
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title = "In situ structure and activity studies of an enzyme adsorbed on spectroscopically undetectable particles",
abstract = "The structural characteristics and the activity of a hyperthermophilic endoglucanase were investigated upon adsorption. Silica (hydrophilic) and Teflon (hydrophobic) surfaces were selected for the study. The materials were specially designed so that the interaction of the particles with light was negligible, and the enzyme conformation in the adsorbed state was monitored in situ. The adsorption isotherms were determined, and the adsorbed endoglucanase was studied using a number of spectroscopic techniques, enzymatic activity tests, and dynamic light scattering. Experiments were performed at pH values below, at, and above the isoelectric point of the enzyme. It was shown that the enzyme adsorbed on the hydrophobic surface of Teflon with higher affinity as compared to the hydrophilic silica nanoparticles. In all cases, adsorption was followed by (slight) changes in the secondary structure resulting in decreased -structural content. The changes were more profound upon adsorption on Teflon. The adsorbed enzyme remained active in the adsorbed state in spite of the structural changes induced when interacting with the surfaces",
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author = "S. Koutsopoulos and A.M. Tjeerdsma and {van Lieshout}, J.F.T. and {van der Oost}, J. and W. Norde",
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In situ structure and activity studies of an enzyme adsorbed on spectroscopically undetectable particles. / Koutsopoulos, S.; Tjeerdsma, A.M.; van Lieshout, J.F.T.; van der Oost, J.; Norde, W.

In: Biomacromolecules, Vol. 6, No. 3, 2005, p. 1176-1184.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - In situ structure and activity studies of an enzyme adsorbed on spectroscopically undetectable particles

AU - Koutsopoulos, S.

AU - Tjeerdsma, A.M.

AU - van Lieshout, J.F.T.

AU - van der Oost, J.

AU - Norde, W.

PY - 2005

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N2 - The structural characteristics and the activity of a hyperthermophilic endoglucanase were investigated upon adsorption. Silica (hydrophilic) and Teflon (hydrophobic) surfaces were selected for the study. The materials were specially designed so that the interaction of the particles with light was negligible, and the enzyme conformation in the adsorbed state was monitored in situ. The adsorption isotherms were determined, and the adsorbed endoglucanase was studied using a number of spectroscopic techniques, enzymatic activity tests, and dynamic light scattering. Experiments were performed at pH values below, at, and above the isoelectric point of the enzyme. It was shown that the enzyme adsorbed on the hydrophobic surface of Teflon with higher affinity as compared to the hydrophilic silica nanoparticles. In all cases, adsorption was followed by (slight) changes in the secondary structure resulting in decreased -structural content. The changes were more profound upon adsorption on Teflon. The adsorbed enzyme remained active in the adsorbed state in spite of the structural changes induced when interacting with the surfaces

AB - The structural characteristics and the activity of a hyperthermophilic endoglucanase were investigated upon adsorption. Silica (hydrophilic) and Teflon (hydrophobic) surfaces were selected for the study. The materials were specially designed so that the interaction of the particles with light was negligible, and the enzyme conformation in the adsorbed state was monitored in situ. The adsorption isotherms were determined, and the adsorbed endoglucanase was studied using a number of spectroscopic techniques, enzymatic activity tests, and dynamic light scattering. Experiments were performed at pH values below, at, and above the isoelectric point of the enzyme. It was shown that the enzyme adsorbed on the hydrophobic surface of Teflon with higher affinity as compared to the hydrophilic silica nanoparticles. In all cases, adsorption was followed by (slight) changes in the secondary structure resulting in decreased -structural content. The changes were more profound upon adsorption on Teflon. The adsorbed enzyme remained active in the adsorbed state in spite of the structural changes induced when interacting with the surfaces

KW - archaeon pyrococcus-furiosus

KW - solid-liquid interfaces

KW - circular-dichroism

KW - proteins

KW - adsorption

KW - endo-beta-1,3-glucanase

KW - extremophiles

KW - surfaces

KW - features

KW - sugar

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