TY - JOUR
T1 - Modulating D-amino acid oxidase (DAAO) substrate specificity through facilitated solvent access
AU - Subramanian, Kalyanasundaram
AU - Góra, Artur
AU - Spruijt, Ruud
AU - Mitusińska, Karolina
AU - Suarez-Diez, Maria
AU - Martins Dos Santos, Vitor
AU - Schaap, Peter J.
PY - 2018/1/1
Y1 - 2018/1/1
N2 - D-amino acid oxidase (DAAO) degrades D-amino acids to produce α-ketoacids, hydrogen peroxide and ammonia. DAAO has often been investigated and engineered for industrial and clinical applications. We combined information from literature with a detailed analysis of the structure to engineer mammalian DAAOs. The structural analysis was complemented with molecular dynamics simulations to characterize solvent accessibility and product release mechanisms. We identified non-obvious residues located on the loops on the border between the active site and the secondary binding pocket essential for pig and human DAAO substrate specificity and activity. We engineered DAAOs by mutating such critical residues and characterised the biochemical activity of the resulting variants. The results highlight the importance of the selected residues in modulating substrate specificity, product egress and enzyme activity, suggesting further steps of DAAO re-engineering towards desired clinical and industrial applications.
AB - D-amino acid oxidase (DAAO) degrades D-amino acids to produce α-ketoacids, hydrogen peroxide and ammonia. DAAO has often been investigated and engineered for industrial and clinical applications. We combined information from literature with a detailed analysis of the structure to engineer mammalian DAAOs. The structural analysis was complemented with molecular dynamics simulations to characterize solvent accessibility and product release mechanisms. We identified non-obvious residues located on the loops on the border between the active site and the secondary binding pocket essential for pig and human DAAO substrate specificity and activity. We engineered DAAOs by mutating such critical residues and characterised the biochemical activity of the resulting variants. The results highlight the importance of the selected residues in modulating substrate specificity, product egress and enzyme activity, suggesting further steps of DAAO re-engineering towards desired clinical and industrial applications.
U2 - 10.1371/journal.pone.0198990
DO - 10.1371/journal.pone.0198990
M3 - Article
C2 - 29906280
AN - SCOPUS:85055206994
SN - 1932-6203
VL - 13
JO - PLoS ONE
JF - PLoS ONE
IS - 6
M1 - e0198990
ER -