Immobilization of the enzyme GpdQ on magnetite nanoparticles for organophosphate pesticide bioremediation

Lena J. Daumann; James A. Larrabee; David Ollis; Gerhard Schenk; Lawrence R. Gahan

doi:10.1016/j.jinorgbio.2013.10.007

Immobilization of the enzyme GpdQ on magnetite nanoparticles for organophosphate pesticide bioremediation

Lena J. Daumann, James A. Larrabee, David Ollis, Gerhard Schenk, Lawrence R. Gahan^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

48 Citations (Scopus)

Abstract

Annually thousands of people die or suffer from organophosphate (pesticide) poisoning. In order to remove these toxic compounds from the environment, the use of enzymes as bioremediators has been proposed. We report here a Ser127Ala mutant based on the enzyme glycerophosphodiesterase (GpdQ) from Enterobacter aerogenes. The mutant, with improved metal binding abilities, has been immobilized using glutaraldehyde on PAMAM dendrimer-modified magnetite nanoparticles. The immobilized system was characterized using elemental analysis as well as infrared, transmission electron and X-ray photoelectron spectroscopies. The amount of GpdQ that was immobilized with the optimized procedure was 1.488 nmol per g MNP. A kinetic assay has been designed to evaluate the activity of the system towards organophosphoester substrates. The specific activity towards BPNPP directly after immobilization was 3.55 μmol mg^{- 1} min^{- 1}, after one week 3.39 μmol mg^{- 1} min^{- 1} and after 120 days 3.36 μmol mg^{- 1} min ^{- 1}, demonstrating that the immobilized enzyme was active for multiple cycles and could be stored on the nanoparticles for a prolonged period.

Original language	English
Pages (from-to)	1-7
Number of pages	7
Journal	Journal of Inorganic Biochemistry
Volume	131
DOIs	https://doi.org/10.1016/j.jinorgbio.2013.10.007
Publication status	Published - 2014

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10.1016/j.jinorgbio.2013.10.007

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title = "Immobilization of the enzyme GpdQ on magnetite nanoparticles for organophosphate pesticide bioremediation",

abstract = "Annually thousands of people die or suffer from organophosphate (pesticide) poisoning. In order to remove these toxic compounds from the environment, the use of enzymes as bioremediators has been proposed. We report here a Ser127Ala mutant based on the enzyme glycerophosphodiesterase (GpdQ) from Enterobacter aerogenes. The mutant, with improved metal binding abilities, has been immobilized using glutaraldehyde on PAMAM dendrimer-modified magnetite nanoparticles. The immobilized system was characterized using elemental analysis as well as infrared, transmission electron and X-ray photoelectron spectroscopies. The amount of GpdQ that was immobilized with the optimized procedure was 1.488 nmol per g MNP. A kinetic assay has been designed to evaluate the activity of the system towards organophosphoester substrates. The specific activity towards BPNPP directly after immobilization was 3.55 μmol mg- 1 min- 1, after one week 3.39 μmol mg- 1 min- 1 and after 120 days 3.36 μmol mg- 1 min - 1, demonstrating that the immobilized enzyme was active for multiple cycles and could be stored on the nanoparticles for a prolonged period.",

keywords = "Bioremediation, Magnetite nanoparticles, Metalloenzyme, Organophosphate pesticide",

author = "Daumann, {Lena J.} and Larrabee, {James A.} and David Ollis and Gerhard Schenk and Gahan, {Lawrence R.}",

year = "2014",

doi = "10.1016/j.jinorgbio.2013.10.007",

language = "English",

volume = "131",

pages = "1--7",

journal = "Journal of Inorganic Biochemistry",

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T1 - Immobilization of the enzyme GpdQ on magnetite nanoparticles for organophosphate pesticide bioremediation

AU - Daumann, Lena J.

AU - Larrabee, James A.

AU - Ollis, David

AU - Schenk, Gerhard

AU - Gahan, Lawrence R.

PY - 2014

Y1 - 2014

N2 - Annually thousands of people die or suffer from organophosphate (pesticide) poisoning. In order to remove these toxic compounds from the environment, the use of enzymes as bioremediators has been proposed. We report here a Ser127Ala mutant based on the enzyme glycerophosphodiesterase (GpdQ) from Enterobacter aerogenes. The mutant, with improved metal binding abilities, has been immobilized using glutaraldehyde on PAMAM dendrimer-modified magnetite nanoparticles. The immobilized system was characterized using elemental analysis as well as infrared, transmission electron and X-ray photoelectron spectroscopies. The amount of GpdQ that was immobilized with the optimized procedure was 1.488 nmol per g MNP. A kinetic assay has been designed to evaluate the activity of the system towards organophosphoester substrates. The specific activity towards BPNPP directly after immobilization was 3.55 μmol mg- 1 min- 1, after one week 3.39 μmol mg- 1 min- 1 and after 120 days 3.36 μmol mg- 1 min - 1, demonstrating that the immobilized enzyme was active for multiple cycles and could be stored on the nanoparticles for a prolonged period.

AB - Annually thousands of people die or suffer from organophosphate (pesticide) poisoning. In order to remove these toxic compounds from the environment, the use of enzymes as bioremediators has been proposed. We report here a Ser127Ala mutant based on the enzyme glycerophosphodiesterase (GpdQ) from Enterobacter aerogenes. The mutant, with improved metal binding abilities, has been immobilized using glutaraldehyde on PAMAM dendrimer-modified magnetite nanoparticles. The immobilized system was characterized using elemental analysis as well as infrared, transmission electron and X-ray photoelectron spectroscopies. The amount of GpdQ that was immobilized with the optimized procedure was 1.488 nmol per g MNP. A kinetic assay has been designed to evaluate the activity of the system towards organophosphoester substrates. The specific activity towards BPNPP directly after immobilization was 3.55 μmol mg- 1 min- 1, after one week 3.39 μmol mg- 1 min- 1 and after 120 days 3.36 μmol mg- 1 min - 1, demonstrating that the immobilized enzyme was active for multiple cycles and could be stored on the nanoparticles for a prolonged period.

KW - Bioremediation

KW - Magnetite nanoparticles

KW - Metalloenzyme

KW - Organophosphate pesticide

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DO - 10.1016/j.jinorgbio.2013.10.007

M3 - Article

SN - 0162-0134

VL - 131

SP - 1

EP - 7

JO - Journal of Inorganic Biochemistry

JF - Journal of Inorganic Biochemistry

ER -

Immobilization of the enzyme GpdQ on magnetite nanoparticles for organophosphate pesticide bioremediation

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