Three in One: Temperature, Solvent and Catalytic Stability by Engineering the Cofactor-Binding Element of Amine Transaminase

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Bibtex

@article{d99f3583b61243fc8ed6e23670d09fdc,
title = "Three in One: Temperature, Solvent and Catalytic Stability by Engineering the Cofactor-Binding Element of Amine Transaminase",
abstract = "Amine transaminase (ATA) catalyse enantioselectively the direct amination of ketones, but insufficient stability during catalysis limits their industrial applicability. Recently, we revealed that ATAs suffer from substrate-induced inactivation mechanism involving dissociation of the enzyme-cofactor intermediate. Here, we report on engineering the cofactor-ring-binding element, which also shapes the active-site entrance. Only two point mutations in this motif improved temperature and catalytic stability in both biphasic media and organic solvent. Thermodynamic analysis revealed a higher melting point for the enzyme-cofactor intermediate. The high cofactor affinity eliminates the need for pyridoxal 5′-phosphate supply, thus making large-scale reactions more cost effective. This is the first report on stabilising a tetrameric ATA by mutating a single structural element. As this structural {"}hotspot{"} is a common feature of other transaminases it could serve as a general engineering target.",
keywords = "Amines, Enzyme catalysis, Operational stability, Pyridoxamine 5′-phosphate, Transaminase",
author = "Tim B{\"o}rner and Sebastian R{\"a}misch and Sebastian Bartsch and Andreas Vogel and Patrick Adlercreutz and Carl Grey",
year = "2017",
month = "8",
day = "4",
doi = "10.1002/cbic.201700236",
language = "English",
volume = "18",
pages = "1482--1486",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "John Wiley & Sons",
number = "15",

}