Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres
| dc.contributor.author | Twala, Busisiwe V | |
| dc.contributor.author | Sewell, Trevor B | |
| dc.contributor.author | Jordaan, Justin | |
| dc.date.accessioned | 2016-09-01T10:59:54Z | |
| dc.date.available | 2016-09-01T10:59:54Z | |
| dc.date.issued | 2012 | |
| dc.date.updated | 2016-09-01T10:58:58Z | |
| dc.description.abstract | The use of enzymes in industrial applications is limited by their instability, cost and difficulty in their recovery and re-use. Immobilisation is a technique which has been shown to alleviate these limitations in biocatalysis. Here we describe the immobilisation of two biocatalytically relevant co-factor recycling enzymes, glucose dehydrogenase (GDH) and NADH oxidase (NOD) on aldehyde functional ReSyn™ polymer microspheres with varying functional group densities. The successful immobilisation of the enzymes on this new high capacity microsphere technology resulted in the maintenance of activity of ∼40% for GDH and a maximum of 15.4% for NOD. The microsphere variant with highest functional group density of ∼3500 μmol g−1 displayed the highest specific activity for the immobilisation of both enzymes at 33.22 U mg−1 and 6.75 U mg−1 for GDH and NOD with respective loading capacities of 51% (0.51 mg mg−1) and 129% (1.29 mg mg−1). The immobilised GDH further displayed improved activity in the acidic pH range. Both enzymes displayed improved pH and thermal stability with the most pronounced thermal stability for GDH displayed on ReSyn™ A during temperature incubation at 65 °C with a 13.59 fold increase, and NOD with a 2.25-fold improvement at 45 °C on the same microsphere variant. An important finding is the suitability of the microspheres for stabilisation of the multimeric protein GDH. | en_ZA |
| dc.identifier | http://dx.doi.org/10.1016/j.enzmictec.2012.03.003 | |
| dc.identifier.apacitation | Twala, B. V., Sewell, T. B., & Jordaan, J. (2012). Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres. <i>Enzyme and Microbial Technology</i>, http://hdl.handle.net/11427/21646 | en_ZA |
| dc.identifier.chicagocitation | Twala, Busisiwe V, Trevor B Sewell, and Justin Jordaan "Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres." <i>Enzyme and Microbial Technology</i> (2012) http://hdl.handle.net/11427/21646 | en_ZA |
| dc.identifier.citation | Twala, B. V., Sewell, B. T., & Jordaan, J. (2012). Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres. Enzyme and microbial technology, 50(6), 331-336. | en_ZA |
| dc.identifier.issn | 0141-0229 | en_ZA |
| dc.identifier.ris | TY - Journal Article AU - Twala, Busisiwe V AU - Sewell, Trevor B AU - Jordaan, Justin AB - The use of enzymes in industrial applications is limited by their instability, cost and difficulty in their recovery and re-use. Immobilisation is a technique which has been shown to alleviate these limitations in biocatalysis. Here we describe the immobilisation of two biocatalytically relevant co-factor recycling enzymes, glucose dehydrogenase (GDH) and NADH oxidase (NOD) on aldehyde functional ReSyn™ polymer microspheres with varying functional group densities. The successful immobilisation of the enzymes on this new high capacity microsphere technology resulted in the maintenance of activity of ∼40% for GDH and a maximum of 15.4% for NOD. The microsphere variant with highest functional group density of ∼3500 μmol g−1 displayed the highest specific activity for the immobilisation of both enzymes at 33.22 U mg−1 and 6.75 U mg−1 for GDH and NOD with respective loading capacities of 51% (0.51 mg mg−1) and 129% (1.29 mg mg−1). The immobilised GDH further displayed improved activity in the acidic pH range. Both enzymes displayed improved pH and thermal stability with the most pronounced thermal stability for GDH displayed on ReSyn™ A during temperature incubation at 65 °C with a 13.59 fold increase, and NOD with a 2.25-fold improvement at 45 °C on the same microsphere variant. An important finding is the suitability of the microspheres for stabilisation of the multimeric protein GDH. DA - 2012 DB - OpenUCT DP - University of Cape Town J1 - Enzyme and Microbial Technology LK - https://open.uct.ac.za PB - University of Cape Town PY - 2012 SM - 0141-0229 T1 - Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres TI - Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres UR - http://hdl.handle.net/11427/21646 ER - | en_ZA |
| dc.identifier.uri | http://hdl.handle.net/11427/21646 | |
| dc.identifier.uri | http://www.sciencedirect.com/science/article/pii/S0141022912000348 | |
| dc.identifier.vancouvercitation | Twala BV, Sewell TB, Jordaan J. Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres. Enzyme and Microbial Technology. 2012; http://hdl.handle.net/11427/21646. | en_ZA |
| dc.language.iso | eng | |
| dc.publisher | Elsevier | en_ZA |
| dc.publisher.institution | University of Cape Town | |
| dc.source | Enzyme and Microbial Technology | en_ZA |
| dc.source.uri | http://www.journals.elsevier.com/enzyme-and-microbial-technology/ | |
| dc.subject.other | Immobilisation | |
| dc.subject.other | Dehydrogenase | |
| dc.subject.other | Oxidase | |
| dc.subject.other | ReSyn™ | |
| dc.subject.other | Stability | |
| dc.subject.other | Biocatalysis | |
| dc.title | Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres | en_ZA |
| dc.type | Journal Article | en_ZA |
| uct.type.filetype | Text | |
| uct.type.filetype | Image | |
| uct.type.publication | Research | en_ZA |
| uct.type.resource | Article | en_ZA |