TY  - JOUR
AU  - Zhu, Z.
AU  - Wang, M.
AU  - Gautam, A.
AU  - Nazor, Jovana
AU  - Momeu, C.
AU  - Prodanović, Radivoje
AU  - Schwaneberg, U.
PY  - 2007
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/77
AB  - A directed evolution protocol was developed for glucose oxidase (GOx) from Aspergillus niger that mimics applications conditions and employs a well-known mediator, oxidized ferrocenemethanol, in a medium throughput screen (96-well plate format). Upon reduction, oxidized ferrocenemethanol shows a color change from blue to pale yellow that can be recorded at 625 nm. Under optimized screening conditions, a CV of less than 20% was achieved in 96-well microtiter plates. For validating the screening system, two mutant libraries of GOx were generated by standard error-prone PCR conditions (0.04 mM MnCl2) and Saccharomyces cerevisiae was employed as host for secreted GOx expression. Two screening of ∼2000 GOx mutants yielded a double mutant (T30S I94V) with improved pH and thermal resistance. Thermal resistance at a residual activity of 50% was increased from 58°C (wild type, WT) to 62°C (T30S I94V) and pH stability was improved at basic pH (pH 8-11). Km for glucose remained nearly unchanged (20.8 mM WT; 21.3 mM T30S I94V) and kcat increased (69.5/s WT; 137.7/s T30S 194V). © 2007 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
T2  - Biotechnology Journal
T1  - Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer
VL  - 2
IS  - 2
SP  - 241
EP  - 248
DO  - 10.1002/biot.200600185
ER  - 

@article{
author = "Zhu, Z. and Wang, M. and Gautam, A. and Nazor, Jovana and Momeu, C. and Prodanović, Radivoje and Schwaneberg, U.",
year = "2007",
abstract = "A directed evolution protocol was developed for glucose oxidase (GOx) from Aspergillus niger that mimics applications conditions and employs a well-known mediator, oxidized ferrocenemethanol, in a medium throughput screen (96-well plate format). Upon reduction, oxidized ferrocenemethanol shows a color change from blue to pale yellow that can be recorded at 625 nm. Under optimized screening conditions, a CV of less than 20% was achieved in 96-well microtiter plates. For validating the screening system, two mutant libraries of GOx were generated by standard error-prone PCR conditions (0.04 mM MnCl2) and Saccharomyces cerevisiae was employed as host for secreted GOx expression. Two screening of ∼2000 GOx mutants yielded a double mutant (T30S I94V) with improved pH and thermal resistance. Thermal resistance at a residual activity of 50% was increased from 58°C (wild type, WT) to 62°C (T30S I94V) and pH stability was improved at basic pH (pH 8-11). Km for glucose remained nearly unchanged (20.8 mM WT; 21.3 mM T30S I94V) and kcat increased (69.5/s WT; 137.7/s T30S 194V). © 2007 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.",
journal = "Biotechnology Journal",
title = "Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer",
volume = "2",
number = "2",
pages = "241-248",
doi = "10.1002/biot.200600185"
}

Zhu, Z., Wang, M., Gautam, A., Nazor, J., Momeu, C., Prodanović, R.,& Schwaneberg, U.. (2007). Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer. in Biotechnology Journal, 2(2), 241-248.
https://doi.org/10.1002/biot.200600185

Zhu Z, Wang M, Gautam A, Nazor J, Momeu C, Prodanović R, Schwaneberg U. Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer. in Biotechnology Journal. 2007;2(2):241-248.
doi:10.1002/biot.200600185 .

Zhu, Z., Wang, M., Gautam, A., Nazor, Jovana, Momeu, C., Prodanović, Radivoje, Schwaneberg, U., "Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer" in Biotechnology Journal, 2, no. 2 (2007):241-248,
https://doi.org/10.1002/biot.200600185 . .