1,5-Pentanediamine (pentanediamine) has good biological activity and is widely used in agriculture, medicine and other industries. Lysine decarboxylase can catalyze L-lysine to produce pentanediamine. In order to improve the efficiency of lysine decarboxylase to catalyze the synthesis of pentanediamine, we first screened, expressed and characterized lysine decarboxylases derived from Pseudomonas putida KT2440, Gluconobacter oxydans DSM3504, Bacillus licheniformis DSM13 and Serratia marcescens (SmcadA), in Escherichia coli. The biochemical characteristics showed that the optimal catalytic pH of SmcadA was 6.0 and the optimal catalytic temperature was about 40 ℃. Subsequently, we conducted a mutation study on the 348th amino acid of SmcadA, screened and obtained a mutant Gly348Ala with significantly improved catalytic efficiency. The main reason was that the mutation caused an increase in the number of hydrogen bond interactions between the amino acid residues of the protein and the substrate lysine, thereby influencing the proteins catalytic efficiency. Finally, the recombinant strain cells were studied for the synthesis of pentanediamine. The recombinant bacteria cells containing the mutant Gly348Ala could synthesize 218.2 g/L pentanediamine, while the wild-type SmcadA recombinant bacteria could only catalyze the synthesis of 159.2 g/L pentanediamine. The results provide a reference for the synthesis of pentanediamine catalyzed by industrialized enzymes.
OSIRE Tolbert
,
YANG Taowei
,
QIAO Zhina
,
SUN Yang
,
XU Meijuan
,
ZHANG Xian
,
SHAO Minglong
,
RAO Zhiming
. Molecular modification of lysine decarboxylase for catalytic synthesis of pentanediamine[J]. Food and Fermentation Industries, 2022
, 48(1)
: 8
-14
.
DOI: 10.13995/j.cnki.11-1802/ts.027293
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