A novel gene AAP that encodes aspartyl aminopeptidase from the peptidase family M18 in Aspergillus oryzae was discovered by bioinformatics analysis. The gene AAP was cloned and overexpressed in Pichia pastoris GS115. Enzymatic properties of recombinant AAP and its application in the hydrolysis of soybean protein were investigated. The results showed that the recombinant AAP had a molecular weight of about 54 kDa. It was active over a broad pH range (6.0-9.5). Its optimal reaction temperature and pH were 50 ℃ and 7.0, respectively. Moreover, Zn2+ and Ca2+ promoted the activity of recombinant AAP, while EDTA significantly inhibited its activity. Using Asp-pNA as a substrate, the Km and Vmax of the enzyme were 0.42 mmol/L and 21.57 μmol/(mL·min), respectively. Recombinant AAP exhibited a substrate specificity of hydrolyzing N-terminal aspartyl and glutamyl residues. In addition, recombinant AAP significantly decreased the bitterness of soybean pepides and improved its hydrolysis degree, which indicated its potential applications in food industries.
SONG Tingting
,
WU Zhenzhen
,
ZONG Hong
,
LU Xinyao
,
ZHU GE Bin
. Heterologous expression of a novel AAP gene from Aspergillus oryzae and characterization of the enzyme[J]. Food and Fermentation Industries, 2019
, 45(5)
: 38
-44
.
DOI: 10.13995/j.cnki.11-1802/ts.018831
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