“Toolbox” construction of an extremophilic nitrile hydratase from Streptomyces thermoautotrophicus for the promising industrial production of various amides

Highly efficient biosynthesis of isonicotinamide through a substrate access tunnel engineered nitrile hydratase from Carbonactinospora thermoautotrophicus

“…bL48D creates a larger active site cavity. 14 We can also observe a better orientation towards the cobalt ion. In the case of bL48D, Co-N distances were much lower than WT (4.67 Å and 5.47 Å vs. 6.13 Å and 5.86 Å).…”

“…In order to achieve highly efficient biosynthesis of isonicotinamide, a thermophilic NHase from Carbonactinospora thermoautotrophicus (C.t NHase) was chosen because it exhibits better stability than the other NHases that have been reported. [14][15][16] The specific activity of C.t NHase was then enhanced using the ''toolbox'' screening strategy established in our previous research. 14 The specific activity of WT and its variants was measured, and the highest activity was exhibited in its corresponding mutant, bL48D, showing a tremendous increase in catalytic activity compared to its parent enzyme (Fig.…”

“…[14][15][16] The specific activity of C.t NHase was then enhanced using the ''toolbox'' screening strategy established in our previous research. 14 The specific activity of WT and its variants was measured, and the highest activity was exhibited in its corresponding mutant, bL48D, showing a tremendous increase in catalytic activity compared to its parent enzyme (Fig. 1).…”

“…S3, ESI †) show that the structure is preserved during the MD simulations and the time evolution is very similar to other studies. 14,15,[18][19][20] Since the heterotetramer model was simulated in each simulation, we could analyze two dimers, each interacting with the ligand, and thus our statistics are twice better.…”

“…In this study, based on the excellent thermostability of C.t NHase, the catalytic activity of C.t NHase was further enhanced by the ''toolbox'' screening strategy established in our previous research. 14 One corresponding mutant, bL48D, showed a tremendous increase in the catalytic activity compared to its parent enzyme. The residue located in the substrate access tunnel of NHase, bLeu48, was found to play a key role in directing the activity of NHase.…”