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A novel combined thermometric and amperometric biosensor for lactose determination based on immobilised cellobiose dehydrogenase.

Yakovleva, Maria LU ; Buzas, Orsolya; Matsumura, Hirotoshi LU ; Samejima, Masahiro; Igarashi, Kiyohiko; Larsson, Per-Olof LU ; Gorton, Lo LU and Danielsson, Bengt LU (2012) In Biosensors & Bioelectronics 31. p.251-256
Abstract
A novel method for lactose determination in milk is proposed. It is based on oxidation of lactose by cellobiose dehydrogenase (CDH) from the basidiomycete Phanerochaete chrysosporium, immobilised in an enzyme reactor. The reactor was prepared by cross-linking CDH onto aminopropyl-silanised controlled pore glass (CPG) beads using glutaraldehyde. The combined biosensor worked in flow injection analysis (FIA) mode and was developed for simultaneous monitoring of the thermometric signal associated with the enzymatic oxidation of lactose using p-benzoquinone as electron acceptor and the electrochemically generated current associated with the oxidation of the hydroquinone formed. A highly reproducible linear response for lactose was obtained... (More)
A novel method for lactose determination in milk is proposed. It is based on oxidation of lactose by cellobiose dehydrogenase (CDH) from the basidiomycete Phanerochaete chrysosporium, immobilised in an enzyme reactor. The reactor was prepared by cross-linking CDH onto aminopropyl-silanised controlled pore glass (CPG) beads using glutaraldehyde. The combined biosensor worked in flow injection analysis (FIA) mode and was developed for simultaneous monitoring of the thermometric signal associated with the enzymatic oxidation of lactose using p-benzoquinone as electron acceptor and the electrochemically generated current associated with the oxidation of the hydroquinone formed. A highly reproducible linear response for lactose was obtained between 0.05mM and 30mM. For a set of more than 500 samples an R.S.D. of less than 10% was achieved. The assay time was ca. 2min per sample. The sensor was applied for the determination of lactose in dairy milk samples (milk with a fat content of 1.5% or 3% and also "lactose free" milk). No sample preparation except dilution with buffer was needed. The proposed method is rapid, suitable for repeated use and allows the possibility to compare results from two different detection methods, thus providing a built-in quality assurance. Some differences in the response observed between the methods indicate that the dual approach can be useful in mechanistic studies of redox enzymes. In addition, a dual system opens up interesting possibilities for studies of enzyme properties and mechanisms. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biosensors & Bioelectronics
volume
31
pages
251 - 256
publisher
Elsevier
external identifiers
  • wos:000300468400039
  • pmid:22078845
  • scopus:84455208197
ISSN
1873-4235
DOI
10.1016/j.bios.2011.10.027
language
English
LU publication?
yes
id
41ed4635-dd55-4eb4-bf94-dcce89b28e88 (old id 2220850)
date added to LUP
2011-12-13 17:32:25
date last changed
2017-09-24 03:55:56
@article{41ed4635-dd55-4eb4-bf94-dcce89b28e88,
  abstract     = {A novel method for lactose determination in milk is proposed. It is based on oxidation of lactose by cellobiose dehydrogenase (CDH) from the basidiomycete Phanerochaete chrysosporium, immobilised in an enzyme reactor. The reactor was prepared by cross-linking CDH onto aminopropyl-silanised controlled pore glass (CPG) beads using glutaraldehyde. The combined biosensor worked in flow injection analysis (FIA) mode and was developed for simultaneous monitoring of the thermometric signal associated with the enzymatic oxidation of lactose using p-benzoquinone as electron acceptor and the electrochemically generated current associated with the oxidation of the hydroquinone formed. A highly reproducible linear response for lactose was obtained between 0.05mM and 30mM. For a set of more than 500 samples an R.S.D. of less than 10% was achieved. The assay time was ca. 2min per sample. The sensor was applied for the determination of lactose in dairy milk samples (milk with a fat content of 1.5% or 3% and also "lactose free" milk). No sample preparation except dilution with buffer was needed. The proposed method is rapid, suitable for repeated use and allows the possibility to compare results from two different detection methods, thus providing a built-in quality assurance. Some differences in the response observed between the methods indicate that the dual approach can be useful in mechanistic studies of redox enzymes. In addition, a dual system opens up interesting possibilities for studies of enzyme properties and mechanisms.},
  author       = {Yakovleva, Maria and Buzas, Orsolya and Matsumura, Hirotoshi and Samejima, Masahiro and Igarashi, Kiyohiko and Larsson, Per-Olof and Gorton, Lo and Danielsson, Bengt},
  issn         = {1873-4235},
  language     = {eng},
  pages        = {251--256},
  publisher    = {Elsevier},
  series       = {Biosensors & Bioelectronics},
  title        = {A novel combined thermometric and amperometric biosensor for lactose determination based on immobilised cellobiose dehydrogenase.},
  url          = {http://dx.doi.org/10.1016/j.bios.2011.10.027},
  volume       = {31},
  year         = {2012},
}