Nitrile hydratase (nitrile hydratase, NHase, EC4.2.1.84) is a kind of metalloenzymes that hydrates nitriles to produce amides. It is used to produce nicotinamide through biological methods in the industry. Due to the exothermic process of the hydration reaction, the industry calls for NHase with higher stability. An NHase gene from thermophilic bacteria Caldalkalibacillus thermarum TA2. A1 was obtained by gene mining in our previous study, however, its catalytic activity was relatively low and cannot meet the application requirements in the industry. In this study, substrate access tunnel engineering was applied and after introducing lysine to βGly42, the specific activity of the βG42K mutant toward 3-cyanopyridine was 2.5 times higher than that of its parent enzyme with minimal loss of stability. The activity of the bacterial cells harboring the βG42K mutant reached 5 539.0 U/mL in the 5 L fermentor, which showed great potential for further application of amide production.
ZHANG Guanglin
,
ZHOU Zhemin
. Enhancing the catalytic performance of thermophilic nitrile hydratase toward 3-cyanopyridine based on substrate access tunnel engineering[J]. Food and Fermentation Industries, 2022
, 48(21)
: 24
-29
.
DOI: 10.13995/j.cnki.11-1802/ts.032826
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