Measurement of adsorption constants of laccase on gold nanoparticles to evaluate the enhancement in enzyme activity of adsorbed laccase

Miguel Peixoto de Almeida, Pedro Quaresma, Susana Sousa, Cláudia Couto, Inês Gomes, Ludwig Krippahl, Ricardo Franco, Eulália Pereira

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Adsorption of enzymes to nanoparticles and the mechanisms responsible for enzyme activity modulation of adsorbed enzymes are not well understood. In this work, gold nanoparticles were used for electrostatic adsorption of a plant-derived laccase. Adsorption constants were determined by four independent techniques: dynamic light scattering, electrophoretic light scattering, agarose gel electrophoresis and fluorescence quenching. Stable bionanoconjugates were formed with logK in the range 6.8-8.9. An increase in enzyme activity was detected, in particular at acidic and close to neutral pH values, a feature that expands the useful pH range of the enzyme. A model for the adsorption was developed, based on geometrical considerations and volume increase data from dynamic light scattering. This indicates that enzymes adsorbed to gold nanoparticles are ca. 9 times more active than the free enzyme.

Original languageEnglish
Pages (from-to)16761-16769
Number of pages9
JournalPhysical Chemistry Chemical Physics
Volume20
Issue number24
DOIs
Publication statusPublished - 1 Jan 2018

Fingerprint

Laccase
enzyme activity
Enzyme activity
Gold
enzymes
gold
Nanoparticles
Adsorption
nanoparticles
adsorption
augmentation
Enzymes
light scattering
Dynamic light scattering
electrophoresis
Electrophoresis
Light scattering
Sepharose
Electrostatics
Quenching

Cite this

@article{7986d92a4dbb450e8eaff6920e8becea,
title = "Measurement of adsorption constants of laccase on gold nanoparticles to evaluate the enhancement in enzyme activity of adsorbed laccase",
abstract = "Adsorption of enzymes to nanoparticles and the mechanisms responsible for enzyme activity modulation of adsorbed enzymes are not well understood. In this work, gold nanoparticles were used for electrostatic adsorption of a plant-derived laccase. Adsorption constants were determined by four independent techniques: dynamic light scattering, electrophoretic light scattering, agarose gel electrophoresis and fluorescence quenching. Stable bionanoconjugates were formed with logK in the range 6.8-8.9. An increase in enzyme activity was detected, in particular at acidic and close to neutral pH values, a feature that expands the useful pH range of the enzyme. A model for the adsorption was developed, based on geometrical considerations and volume increase data from dynamic light scattering. This indicates that enzymes adsorbed to gold nanoparticles are ca. 9 times more active than the free enzyme.",
author = "Almeida, {Miguel Peixoto de} and Pedro Quaresma and Susana Sousa and Cl{\'a}udia Couto and In{\^e}s Gomes and Ludwig Krippahl and Ricardo Franco and Eul{\'a}lia Pereira",
note = "Marta Giza is acknowledged for expert utilization of the UCSF Chimera package. Chimera is developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIGMS P41-GM103311). This work was supported by (a) Unidade de Ciencias Biomoleculares Aplicadas-UCIBIO which is financed by Portuguese national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728); and NOVA LINCS UID/CEC/04516/2013; and (b) European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundacao para a Ciencia e Tecnologia), under the Partnership Agreement PT2020 through project UID/QUI/50006/2013-POCI/01/0145/FEDER/007265 (LAQV/REQUIMTE), Programa Operacional Regional do Norte (ON.2 - O Novo Norte), under the Quadro de Referencia Estrategico Nacional (QREN) and funded by Fundo Europeu de Desenvolvimento Regional NORTE-01-0145-FEDER-000011. PQ, SS and IG acknowledge their postdoctoral fellowships (SFRH/BPD/84018/2012, SFRH/BPD/103788/2014 and SFRH/BPD/63850/2009, respectively), and MPA acknowledges his doctoral fellowship (SFRH/BD/95983/2013); all fellowships are financed by Fundacao para a Ciencia e a Tecnologia, Portugal.",
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Measurement of adsorption constants of laccase on gold nanoparticles to evaluate the enhancement in enzyme activity of adsorbed laccase. / Almeida, Miguel Peixoto de ; Quaresma, Pedro; Sousa, Susana; Couto, Cláudia; Gomes, Inês; Krippahl, Ludwig; Franco, Ricardo; Pereira, Eulália.

In: Physical Chemistry Chemical Physics, Vol. 20, No. 24, 01.01.2018, p. 16761-16769.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Measurement of adsorption constants of laccase on gold nanoparticles to evaluate the enhancement in enzyme activity of adsorbed laccase

AU - Almeida, Miguel Peixoto de

AU - Quaresma, Pedro

AU - Sousa, Susana

AU - Couto, Cláudia

AU - Gomes, Inês

AU - Krippahl, Ludwig

AU - Franco, Ricardo

AU - Pereira, Eulália

N1 - Marta Giza is acknowledged for expert utilization of the UCSF Chimera package. Chimera is developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIGMS P41-GM103311). This work was supported by (a) Unidade de Ciencias Biomoleculares Aplicadas-UCIBIO which is financed by Portuguese national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728); and NOVA LINCS UID/CEC/04516/2013; and (b) European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundacao para a Ciencia e Tecnologia), under the Partnership Agreement PT2020 through project UID/QUI/50006/2013-POCI/01/0145/FEDER/007265 (LAQV/REQUIMTE), Programa Operacional Regional do Norte (ON.2 - O Novo Norte), under the Quadro de Referencia Estrategico Nacional (QREN) and funded by Fundo Europeu de Desenvolvimento Regional NORTE-01-0145-FEDER-000011. PQ, SS and IG acknowledge their postdoctoral fellowships (SFRH/BPD/84018/2012, SFRH/BPD/103788/2014 and SFRH/BPD/63850/2009, respectively), and MPA acknowledges his doctoral fellowship (SFRH/BD/95983/2013); all fellowships are financed by Fundacao para a Ciencia e a Tecnologia, Portugal.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Adsorption of enzymes to nanoparticles and the mechanisms responsible for enzyme activity modulation of adsorbed enzymes are not well understood. In this work, gold nanoparticles were used for electrostatic adsorption of a plant-derived laccase. Adsorption constants were determined by four independent techniques: dynamic light scattering, electrophoretic light scattering, agarose gel electrophoresis and fluorescence quenching. Stable bionanoconjugates were formed with logK in the range 6.8-8.9. An increase in enzyme activity was detected, in particular at acidic and close to neutral pH values, a feature that expands the useful pH range of the enzyme. A model for the adsorption was developed, based on geometrical considerations and volume increase data from dynamic light scattering. This indicates that enzymes adsorbed to gold nanoparticles are ca. 9 times more active than the free enzyme.

AB - Adsorption of enzymes to nanoparticles and the mechanisms responsible for enzyme activity modulation of adsorbed enzymes are not well understood. In this work, gold nanoparticles were used for electrostatic adsorption of a plant-derived laccase. Adsorption constants were determined by four independent techniques: dynamic light scattering, electrophoretic light scattering, agarose gel electrophoresis and fluorescence quenching. Stable bionanoconjugates were formed with logK in the range 6.8-8.9. An increase in enzyme activity was detected, in particular at acidic and close to neutral pH values, a feature that expands the useful pH range of the enzyme. A model for the adsorption was developed, based on geometrical considerations and volume increase data from dynamic light scattering. This indicates that enzymes adsorbed to gold nanoparticles are ca. 9 times more active than the free enzyme.

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U2 - 10.1039/c8cp03116a

DO - 10.1039/c8cp03116a

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JO - Physical Chemistry Chemical Physics

JF - Physical Chemistry Chemical Physics

SN - 1463-9076

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