Fine-tuned preparation of cross-linked laccase nanoaggregates
| dc.authorid | SAHUTOGLU, Arif Sercan/0000-0003-1183-3553 | |
| dc.contributor.author | Sahutoglu, Arif Sercan | |
| dc.contributor.author | Akgul, Cahit | |
| dc.date.accessioned | 2025-01-27T20:16:54Z | |
| dc.date.available | 2025-01-27T20:16:54Z | |
| dc.date.issued | 2019 | |
| dc.department | Çanakkale Onsekiz Mart Üniversitesi | |
| dc.description.abstract | This study focuses on well-known but commonly overlooked or unreported factors in the preparation of cross-linked enzyme nano-aggregates (nano-CLEAs). The parameters including the ionic strength of the protein solution, protein, precipitant and cross-linker concentrations, pH and addition order of the reagents were fine-tuned for nanoaggregate preparation without the need of non-protein support material, special equipment or sophisticated procedures. For this purpose, precipitation as nano-aggregates and then cross-linking while maintaining submicron size distribution were studied independently. Moreover, nano-aggregate formation from reverse micellar solutions was also investigated to improve the scope of the method to membrane-bound enzymes. Five different precipitation agents together with three different cross-linkers were investigated for immobilization of the Trametes versicolor laccase as cross-linked nano-aggregates. Although complete activity recovery was possible for micro-aggregates, the best activity results for nano-aggregates were 40.5%+/- 5.0. The K-m values of the immobilized enzymes were slightly lower than the K-m values of the free counterparts which indicate little or no mass transfer limitation due to the nano-immobilization process. However, V-max values were also lower. The activity loss and V-max reduction upon immobilization were found to mainly result from the modification of the amine groups instead of excess crosslinking. The thermal stabilities of the crosslinked laccase nano-aggregates were significantly higher (similar to 10-30 fold at 60 degrees C) compared to free laccase and the nano-CLEAs retained up to 30% of their initial activities upon 7 consequent usages. The sizes of the obtained immobilized enzyme products were found to be greatly variable depending on the cross-linker type. The smaller particles (similar to 200 nm radius) were obtained when EDAC was used as the cross-linker. The larger products (similar to 600 nm radius) were prepared when the cross-linker was dextran poly-aldehyde. The addition order of the reagents was found to be effective on particle size and thermal stability values. | |
| dc.description.sponsorship | Scientific and Technological Research Council of Turkey (TUBITAK) [215Z097] | |
| dc.description.sponsorship | This work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK) under grant number 215Z097. | |
| dc.identifier.doi | 10.1080/10242422.2019.1602610 | |
| dc.identifier.endpage | 447 | |
| dc.identifier.issn | 1024-2422 | |
| dc.identifier.issn | 1029-2446 | |
| dc.identifier.issue | 6 | |
| dc.identifier.scopus | 2-s2.0-85065543382 | |
| dc.identifier.scopusquality | Q2 | |
| dc.identifier.startpage | 431 | |
| dc.identifier.uri | https://doi.org/10.1080/10242422.2019.1602610 | |
| dc.identifier.uri | https://hdl.handle.net/20.500.12428/21430 | |
| dc.identifier.volume | 37 | |
| dc.identifier.wos | WOS:000471560200001 | |
| dc.identifier.wosquality | Q3 | |
| dc.indekslendigikaynak | Web of Science | |
| dc.indekslendigikaynak | Scopus | |
| dc.language.iso | en | |
| dc.publisher | Taylor & Francis Ltd | |
| dc.relation.ispartof | Biocatalysis and Biotransformation | |
| dc.relation.publicationcategory | info:eu-repo/semantics/openAccess | |
| dc.rights | info:eu-repo/semantics/closedAccess | |
| dc.snmz | KA_WoS_20250125 | |
| dc.subject | Laccase | |
| dc.subject | cross-linked enzyme aggregates | |
| dc.subject | micro-emulsions | |
| dc.subject | nano-immobilization | |
| dc.title | Fine-tuned preparation of cross-linked laccase nanoaggregates | |
| dc.type | Article |
