Artikel ; Online: YESS 2.0, a Tunable Platform for Enzyme Evolution, Yields Highly Active TEV Protease Variants.
ACS synthetic biology
2021 Band 10, Heft 1, Seite(n) 63–71
Abstract: Here we describe YESS 2.0, a highly versatile version of the yeast endoplasmic sequestration screening (YESS) system suitable for engineering and characterizing protein/peptide modifying enzymes such as proteases with desired new activities. By ... ...
Abstract | Here we describe YESS 2.0, a highly versatile version of the yeast endoplasmic sequestration screening (YESS) system suitable for engineering and characterizing protein/peptide modifying enzymes such as proteases with desired new activities. By incorporating features that modulate gene transcription as well as substrate and enzyme spatial sequestration, YESS 2.0 achieves a significantly higher operational and dynamic range compared with the original YESS. To showcase the new advantages of YESS 2.0, we improved an already efficient TEV protease variant (TEV-EAV) to obtain a variant (eTEV) with a 2.25-fold higher catalytic efficiency, derived almost entirely from an increase in turnover rate ( |
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Mesh-Begriff(e) | Amino Acid Sequence ; Biocatalysis ; Endopeptidases/genetics ; Endopeptidases/metabolism ; Evolution, Molecular ; Kinetics ; Metabolic Engineering ; Mutagenesis, Site-Directed ; Peptides/chemistry ; Peptides/metabolism ; Plasmids/genetics ; Plasmids/metabolism ; Saccharomyces cerevisiae/metabolism ; Substrate Specificity |
Chemische Substanzen | Peptides ; Endopeptidases (EC 3.4.-) ; TEV protease (EC 3.4.-) |
Sprache | Englisch |
Erscheinungsdatum | 2021-01-05 |
Erscheinungsland | United States |
Dokumenttyp | Journal Article |
ISSN | 2161-5063 |
ISSN (online) | 2161-5063 |
DOI | 10.1021/acssynbio.0c00452 |
Datenquelle | MEDical Literature Analysis and Retrieval System OnLINE |
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