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

Research output: Contribution to journalArticle


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.


External organisations
  • Nestlé
  • Scripps Research Institute
  • c-LEcta GmbH
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Organic Chemistry


  • Amines, Enzyme catalysis, Operational stability, Pyridoxamine 5′-phosphate, Transaminase
Original languageEnglish
Pages (from-to)1482-1486
Issue number15
Early online date2017 Jun 13
Publication statusPublished - 2017 Aug 4
Publication categoryResearch