| Author | Trevizano, Larissa Mattos | |
| Author | Ventorim, Rafaela Zandonade | |
| Author | Rezende, Sebastião Tavares de | |
| Author | Silva Junior, Floriano Paes | |
| Author | Guimarães, Valéria Monteze | |
| Access date | 2016-08-25T14:40:42Z | |
| Available date | 2016-08-25T14:40:42Z | |
| Document date | 2012 | |
| Citation | TREVIZANO, Larissa Mattos; et al. Thermostability improvement of Orpinomyces sp. xylanase by directed evolution. Journal of Molecular Catalysis B: Enzymatic, v.81, p.12-18, Sept. 2012. | pt_BR |
| ISSN | 1381-1177 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/15399 | |
| Language | eng | pt_BR |
| Publisher | ScienceDirect | pt_BR |
| Rights | restricted access | |
| Subject in Portuguese | Reação em Cadeia da Polimerase | pt_BR |
| Subject in Portuguese | Xylanase | pt_BR |
| Subject in Portuguese | Termoestablidade | pt_BR |
| Subject in Portuguese | Neocallimastigales | pt_BR |
| Title | Thermostability improvement of Orpinomyces sp. xylanase by directed evolution | pt_BR |
| Type | Article | |
| DOI | 10.1016/j.molcatb.2012.04.021 | |
| Abstract | The methodology of directed evolution, using the mutagenic technique of error-prone PCR has been used to improve the thermostability of enzymes. This method was applied to the endo-β-1,4-xylanase from Orpinomyces strain PC-2. The constructed library of xylanase (xynA) mutants was subjected to several screening cycles in plates with azo-xylan-agarose as substrate and four thermostable mutants (M1–M4) were selected. Homology models for these thermostable mutants were constructed to identify the location of the residues changed by error-prone PCR and to investigate the effect of these mutations on the xylanase properties. Xylanase activities of the mutants and wild type were maximal at 60 °C and in the pH range of 5–7. The mutants displayed higher thermostability than the wild type XynA, where the wild type showed a half-life at 60 °C of 7.92 min, while half-life values for M1, M2, M3 and M4 were 209, 33.2, 401 and 15.3 min, respectively. Additionally, M3 and M4 presented a good performance in more extreme pH conditions. The mutants retained their ability to hydrolyze birchwood and oat spelt xylans, which are substrates presenting different degrees of branching. | pt_BR |
| Affilliation | Universidade Federal de Viçosa. BIOAGRO, Viçosa, MG, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Viçosa. BIOAGRO, Viçosa, MG, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Viçosa. BIOAGRO, Viçosa, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz., Instituto Oswaldo Cruz. Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Viçosa. BIOAGRO, Viçosa, MG, Brasil. | pt_BR |
| Subject | Error-prone PCR | pt_BR |
| Subject | Xylanase | pt_BR |
| Subject | Thermostability | pt_BR |
| Subject | Orpinomyces | pt_BR |
| Embargo date | 2030-01-01 | |
