Utilize este identificador para referenciar este registo:
https://hdl.handle.net/1822/74378
Título: | Evolutionary engineering reveals amino acid substitutions in Ato2 and Ato3 that allow improved growth of Saccharomyces cerevisiae on lactic acid |
Autor(es): | Baldi, Nicolò de Valk, Sophie Claire Sousa-Silva, M. Casal, Margarida Soares-Silva, Isabel João Mans, Robert |
Palavras-chave: | Yeast Carboxylate transporters Biotechnology Evolutionary engineering Transport Protein structure Reverse engineering Carboxylic acids |
Data: | Mai-2021 |
Editora: | Federation of European Microbiological Societies |
Revista: | FEMS Yeast Research |
Citação: | Baldi, N., de Valk, S. C., Sousa-Silva, M., Casal, M., Soares-Silva, I., & Mans, R. (2021). Evolutionary engineering reveals amino acid substitutions in Ato2 and Ato3 that allow improved growth of Saccharomyces cerevisiae on lactic acid. FEMS Yeast Research, 21(4) |
Resumo(s): | In Saccharomyces cerevisiae, the complete set of proteins involved in transport of lactic acid across the cell membrane has not been determined. In this study we aimed to identify transport proteins not previously described to be involved in lactic acid transport via a combination of directed evolution, whole-genome resequencing and reverse engineering. Evolution of a strain lacking all known lactic acid transporters on lactate led to the discovery of mutated Ato2 and Ato3 as two novel lactic acid transport proteins. When compared to previously identified S. cerevisiae genes involved in lactic acid transport, expression of ATO3T284C was able to facilitate the highest growth rate (0.15 ± 0.01 h-1) on this carbon source. A comparison between (evolved) sequences and 3D models of the transport proteins showed that most of the identified mutations resulted in a widening of the narrowest hydrophobic constriction of the anion channel. We hypothesize that this observation, sometimes in combination with an increased binding affinity of lactic acid to the sites adjacent to this constriction, are responsible for the improved lactic acid transport in the evolved proteins. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/74378 |
DOI: | 10.1093/femsyr/foab033 |
ISSN: | 1567-1356 |
e-ISSN: | 1567-1364 |
Versão da editora: | https://academic.oup.com/femsyr/article-abstract/21/4/foab033/6286924 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
Aparece nas coleções: | CBMA - Artigos/Papers |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
---|---|---|---|---|
foab033_Vfinal artigo.pdf | 461,8 kB | Adobe PDF | Ver/Abrir |