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Engineering of Cellobiose Dehydrogenases for Improved Glucose Sensitivity and Reduced Maltose Affinityydrogenases for Improved Glucose Sensitivity and Reduced Maltose Affinity

Ortiz, Roberto LU ; Rahman, Mahbubur ; Zangrilli, Beatrice ; Sygmund, Christoph ; Micheelsen, Pernille O. ; Silow, Maria ; Toscano, Miguel D ; Ludwig, Roland and Gorton, Lo LU (2017) In ChemElectroChem 4(4). p.846-855
Abstract

Cellobiose dehydrogenase (CDH) is a fungal extracellular flavocytochrome capable of direct electron transfer (DET). Unlike other CDHs, the pH optimum for CDHs from Corynascus thermophilus (CtCDH) and Humicola insolens (HiCDH) is close to the human physiological pH in blood (7.4). These are, therefore, interesting candidates for glucose measurements in human blood and the application in enzymatic fuel cells is, however, limited by their relatively low activity with this substrate. In this work, the substrate specificities of CtCDH and HiCDH have been altered by a single cysteine to tyrosine substitution in the active sites of CtCDH (position 291) and HiCDH (position 285), which resulted in improved kinetic constants with glucose while... (More)

Cellobiose dehydrogenase (CDH) is a fungal extracellular flavocytochrome capable of direct electron transfer (DET). Unlike other CDHs, the pH optimum for CDHs from Corynascus thermophilus (CtCDH) and Humicola insolens (HiCDH) is close to the human physiological pH in blood (7.4). These are, therefore, interesting candidates for glucose measurements in human blood and the application in enzymatic fuel cells is, however, limited by their relatively low activity with this substrate. In this work, the substrate specificities of CtCDH and HiCDH have been altered by a single cysteine to tyrosine substitution in the active sites of CtCDH (position 291) and HiCDH (position 285), which resulted in improved kinetic constants with glucose while decreasing the activity with several disaccharides, including maltose. The DET properties of the generated CDH variants were tested in the absence and in the presence of substrates, on graphite electrodes and thiolic self-assembled monolayer (SAM)-modified Au electrodes. Seven different thiols with different spacer lengths were used, containing -COOH, -OH, and -NH2 end groups. The length and head functionality of the thiol govern the efficiency of the DET reaction and indicate different DET properties of CtCDH and HiCDH

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author
; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cellobiose dehydrogenase, Direct electron transfer, Enzymatic fuel cells, Glucose sensors, Site-directed mutagenesis
in
ChemElectroChem
volume
4
issue
4
pages
846 - 855
publisher
Wiley-Blackwell
external identifiers
  • wos:000399641500009
  • scopus:85011357375
ISSN
2196-0216
DOI
10.1002/celc.201600781
language
English
LU publication?
yes
id
45414310-1811-49e1-ba2c-66541c67db08
date added to LUP
2017-02-16 07:54:01
date last changed
2025-01-06 07:07:47
@article{45414310-1811-49e1-ba2c-66541c67db08,
  abstract     = {{<p>Cellobiose dehydrogenase (CDH) is a fungal extracellular flavocytochrome capable of direct electron transfer (DET). Unlike other CDHs, the pH optimum for CDHs from Corynascus thermophilus (CtCDH) and Humicola insolens (HiCDH) is close to the human physiological pH in blood (7.4). These are, therefore, interesting candidates for glucose measurements in human blood and the application in enzymatic fuel cells is, however, limited by their relatively low activity with this substrate. In this work, the substrate specificities of CtCDH and HiCDH have been altered by a single cysteine to tyrosine substitution in the active sites of CtCDH (position 291) and HiCDH (position 285), which resulted in improved kinetic constants with glucose while decreasing the activity with several disaccharides, including maltose. The DET properties of the generated CDH variants were tested in the absence and in the presence of substrates, on graphite electrodes and thiolic self-assembled monolayer (SAM)-modified Au electrodes. Seven different thiols with different spacer lengths were used, containing -COOH, -OH, and -NH<sub>2</sub> end groups. The length and head functionality of the thiol govern the efficiency of the DET reaction and indicate different DET properties of CtCDH and HiCDH</p>}},
  author       = {{Ortiz, Roberto and Rahman, Mahbubur and Zangrilli, Beatrice and Sygmund, Christoph and Micheelsen, Pernille O. and Silow, Maria and Toscano, Miguel D and Ludwig, Roland and Gorton, Lo}},
  issn         = {{2196-0216}},
  keywords     = {{Cellobiose dehydrogenase; Direct electron transfer; Enzymatic fuel cells; Glucose sensors; Site-directed mutagenesis}},
  language     = {{eng}},
  number       = {{4}},
  pages        = {{846--855}},
  publisher    = {{Wiley-Blackwell}},
  series       = {{ChemElectroChem}},
  title        = {{Engineering of Cellobiose Dehydrogenases for Improved Glucose Sensitivity and Reduced Maltose Affinityydrogenases for Improved Glucose Sensitivity and Reduced Maltose Affinity}},
  url          = {{http://dx.doi.org/10.1002/celc.201600781}},
  doi          = {{10.1002/celc.201600781}},
  volume       = {{4}},
  year         = {{2017}},
}