Homoserine dehydrogenase (HSD) is a key enzyme in the biosynthesis of aspartate-family amino acids such as L-homoserine and L-threonine.But HSD exhibited low activity and is feedback inhibited by L-threonine, which severely restricts the biosynthesis level of L-homoserine and L-threonine.In this study, eight HSDs from different species were mined through database search.Among of them, BdHSD derived from Brachypodium distachyon had the highest catalytic activity with 7.6 U/mg, and was not feedback-inhibited by L-threonine.The optimal catalytic pH of BdHSD was 10.5, and the optimal catalytic temperature was 38 ℃.To improve the catalytic activity of BdHSD, this study further performed the directed evolution of BdHSD, and three BdHSD mutants T186A, N283K, and A137T/I188V with higher catalytic activity were obtained through multiple rounds of screening.The enzyme activity of the mutant T186A reached 10.3 U/mg, which was 35.6% higher than that of the wild type.The L-homoserine fermentation analysis suggested that the BdHSD mutant could effectively enhance the synthesis level of L-homoserine.In summary, this study had mined and evolved the BdHSD with high efficient catalytic, which provided a powerful catalytic element for the efficient biosynthesis of L-homoserine, L-threonine, L-methionine, and other aspartate-family amino acids.
WU Shuo
,
HUANG Xinyan
,
LI Mengya
,
XU Ning
,
WEI Liang
,
LIU Jun
. Research on mining and evolution of a novel homoserine dehydrogenase[J]. Food and Fermentation Industries, 2024
, 50(12)
: 9
-16
.
DOI: 10.13995/j.cnki.11-1802/ts.035737
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