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Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer
dc.creator | Zhu, Z. | |
dc.creator | Wang, M. | |
dc.creator | Gautam, A. | |
dc.creator | Nazor, Jovana | |
dc.creator | Momeu, C. | |
dc.creator | Prodanović, Radivoje | |
dc.creator | Schwaneberg, U. | |
dc.date.accessioned | 2018-11-22T00:10:24Z | |
dc.date.available | 2018-11-22T00:10:24Z | |
dc.date.issued | 2007 | |
dc.identifier.issn | 1860-6768 | |
dc.identifier.uri | https://cherry.chem.bg.ac.rs/handle/123456789/77 | |
dc.description.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. | en |
dc.rights | restrictedAccess | |
dc.source | Biotechnology Journal | |
dc.subject | Biofuel cell | en |
dc.subject | Directed evolution | en |
dc.subject | Ferrocenemethanol | en |
dc.subject | Glucose oxidase | en |
dc.subject | Mediator | en |
dc.title | Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer | en |
dc.type | article | |
dc.rights.license | ARR | |
dc.citation.volume | 2 | |
dc.citation.issue | 2 | |
dc.citation.spage | 241 | |
dc.citation.epage | 248 | |
dc.identifier.doi | 10.1002/biot.200600185 | |
dc.citation.other | 2(2): 241-248 | |
dc.type.version | publishedVersion | en |
dc.identifier.scopus | 2-s2.0-34248349964 |