Lagerman_2023_Chem.Eng.Sci_277_118804

Reference

Title : Total turnover number prediction of an aggregating biocatalyst: Amino ester hydrolase (AEH) - Lagerman_2023_Chem.Eng.Sci_277_118804
Author(s) : Lagerman CE , Blum JK , Rogers TA , Grover MA , Rousseau RW , Bommarius AS
Ref : Chem Eng Sci , 277 :118804 , 2023
Abstract :

Amino ester hydrolase (AEH) from Xanthomonas campestris is a promising candidate for beta-lactam synthesis but suffers from low thermostability and rapid deactivation. Wild-type AEH was found to deactivate with an apparent order that varies with temperature from n = 2 at 30 C (close to the temperature of optimum activity Topt) to n = 1.5 at 25 C. AEH features two transitional temperatures: (1) from native N to partially unfolded, inactive intermediate I, and (2) from partially unfolded to fully unfolded entity U at the melting temperature Tm. CD and light scattering data suggest aggregation near Topt. To determine the total turnover number TTN, AEH was deactivated by imposing a temperature gradient and recording instantaneous rates of cephalexin hydrolysis. The TTN increased ~ 5-fold from the wild-type (WT) for the quadruple variant N186D/A275P/E143H/V622I (QVH) at 25 C and 10 nM AEH but varied significantly with AEH concentration and temperature.

PubMedSearch : Lagerman_2023_Chem.Eng.Sci_277_118804
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Lagerman CE, Blum JK, Rogers TA, Grover MA, Rousseau RW, Bommarius AS (2023)
Total turnover number prediction of an aggregating biocatalyst: Amino ester hydrolase (AEH)
Chem Eng Sci 277 :118804

Lagerman CE, Blum JK, Rogers TA, Grover MA, Rousseau RW, Bommarius AS (2023)
Chem Eng Sci 277 :118804