Modulating D-amino acid oxidase (DAAO) substrate specificity through facilitated solvent access

Kalyanasundaram Subramanian, Artur Góra*, Ruud Spruijt, Karolina Mitusińska, Maria Suarez-Diez, Vitor Martins Dos Santos, Peter J. Schaap

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

17 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article numbere0198990
JournalPLoS ONE
Volume13
Issue number6
DOIs
Publication statusPublished - 1 Jan 2018

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