Lagerman_2023_Protein.J__

Reference

Title : Improvement of alpha-amino Ester Hydrolase Stability via Computational Protein Design - Lagerman_2023_Protein.J__
Author(s) : Lagerman CE , Joe EA , Grover MA , Rousseau RW , Bommarius AS
Ref : Protein J , : , 2023
Abstract :

Amino ester hydrolases (AEHs) are capable of rapid synthesis of cephalexin but suffer from rapid deactivation even at low temperatures. Previous efforts to engineer AEH have generated several improved variants but have been limited in scope in part due to limitations in activity assay throughput for beta-lactam synthesis reactions. Rational design of 'whole variants' was explored to rapidly improve AEH thermostability by mutating between 3-15% of residues. Most variants were found to be inactive due to a mutated calcium binding site, the function of which has not previously been described. Four active variants, all with improved melting temperatures, were characterized in terms of synthesis and hydrolysis activity, melting temperature, and deactivation at 25 degreesC. Two variants were found to have improved total turnover numbers relative to the initial AEH variant; however, a clear tradeoff exists between improved stability and overall activity of each variant.

PubMedSearch : Lagerman_2023_Protein.J__
PubMedID: 37819423

Related information

Citations formats

Lagerman CE, Joe EA, Grover MA, Rousseau RW, Bommarius AS (2023)
Improvement of alpha-amino Ester Hydrolase Stability via Computational Protein Design
Protein J :

Lagerman CE, Joe EA, Grover MA, Rousseau RW, Bommarius AS (2023)
Protein J :