The University of Southampton
University of Southampton Institutional Repository

Oxidation of NADH produced by a lactate dehydrogenase immobilised on poly(aniline)-poly(anion) composite films

Oxidation of NADH produced by a lactate dehydrogenase immobilised on poly(aniline)-poly(anion) composite films
Oxidation of NADH produced by a lactate dehydrogenase immobilised on poly(aniline)-poly(anion) composite films
The immobilisation of enzymes is important for applications in bioelectrochemistry such as biosensors or biofuel cells. In this paper we report the immobilisation of lactate dehydrogenase (LDH) on poly(aniline)–poly(acrylate) [PANi–PAA] and poly(aniline)–poly(vinylsulfonate) [PANi–PVS] composite films. Two genetically engineered forms of LDH (E.C.1.1.1.27) from Bacillus stearothermophilus, one with a poly(histidine) tag on the C-Terminus (LDH-CHis) the other with a poly(histidine) tag on the N-terminus (LDH-NHis), together with the wild type enzyme (WT-LDH) were studied. The LDH-CHis and LDH-NHis both have better affinity for the poly(aniline)–poly(anion) composite films than the WT-LDH. The immobilised LDH reduces the coenzyme NAD+ to NADH and oxidises the substrate, -lactate, to pyruvate. The NADH produced is then oxidised at the poly(aniline)–poly(anion) composite films. The effects of buffer concentration, temperature, NAD+ concentration, enzyme immobilisation conditions, film thickness and electrode rotation rate on the catalytic current were all investigated.
biosensors, chemically modified electrode, lactate dehydrogenase, adenine-dinucleotide nadh, poly(aniline)-coated electrodes, polyanilinefilms, immobilization, oxidase
1572-6657
253-259
Simon, Evelyne
890fd27c-8885-4929-872f-0afc8d076526
Halliwell, Catherine M.
6cc766e4-a027-4380-8604-c91a27301774
Toh, Chee Seng
87a356a3-63ee-41f6-8aab-84813ad8e30e
Cass, Anthony E.G.
6696e860-4ee0-42ea-958f-2cc9ca999e91
Bartlett, Philip N.
d99446db-a59d-4f89-96eb-f64b5d8bb075
Simon, Evelyne
890fd27c-8885-4929-872f-0afc8d076526
Halliwell, Catherine M.
6cc766e4-a027-4380-8604-c91a27301774
Toh, Chee Seng
87a356a3-63ee-41f6-8aab-84813ad8e30e
Cass, Anthony E.G.
6696e860-4ee0-42ea-958f-2cc9ca999e91
Bartlett, Philip N.
d99446db-a59d-4f89-96eb-f64b5d8bb075

Simon, Evelyne, Halliwell, Catherine M., Toh, Chee Seng, Cass, Anthony E.G. and Bartlett, Philip N. (2002) Oxidation of NADH produced by a lactate dehydrogenase immobilised on poly(aniline)-poly(anion) composite films. Journal of Electroanalytical Chemistry, 538-539, 253-259. (doi:10.1016/S0022-0728(02)00833-1).

Record type: Article

Abstract

The immobilisation of enzymes is important for applications in bioelectrochemistry such as biosensors or biofuel cells. In this paper we report the immobilisation of lactate dehydrogenase (LDH) on poly(aniline)–poly(acrylate) [PANi–PAA] and poly(aniline)–poly(vinylsulfonate) [PANi–PVS] composite films. Two genetically engineered forms of LDH (E.C.1.1.1.27) from Bacillus stearothermophilus, one with a poly(histidine) tag on the C-Terminus (LDH-CHis) the other with a poly(histidine) tag on the N-terminus (LDH-NHis), together with the wild type enzyme (WT-LDH) were studied. The LDH-CHis and LDH-NHis both have better affinity for the poly(aniline)–poly(anion) composite films than the WT-LDH. The immobilised LDH reduces the coenzyme NAD+ to NADH and oxidises the substrate, -lactate, to pyruvate. The NADH produced is then oxidised at the poly(aniline)–poly(anion) composite films. The effects of buffer concentration, temperature, NAD+ concentration, enzyme immobilisation conditions, film thickness and electrode rotation rate on the catalytic current were all investigated.

This record has no associated files available for download.

More information

Published date: 13 December 2002
Keywords: biosensors, chemically modified electrode, lactate dehydrogenase, adenine-dinucleotide nadh, poly(aniline)-coated electrodes, polyanilinefilms, immobilization, oxidase

Identifiers

Local EPrints ID: 19853
URI: http://eprints.soton.ac.uk/id/eprint/19853
ISSN: 1572-6657
PURE UUID: 5f4f5975-a0c1-484d-96a6-c790f7166557
ORCID for Philip N. Bartlett: ORCID iD orcid.org/0000-0002-7300-6900

Catalogue record

Date deposited: 22 Feb 2006
Last modified: 08 Jan 2022 02:38

Export record

Altmetrics

Contributors

Author: Evelyne Simon
Author: Catherine M. Halliwell
Author: Chee Seng Toh
Author: Anthony E.G. Cass

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×