Production and characterization of a novel aminopeptidase A from Lactococcus lactis / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 3494-3507, 2023.
Article
in Chinese
| WPRIM
| ID: wpr-1007972
ABSTRACT
Aminopeptidase A (Pep A) is a metal-dependent enzyme that specifically hydrolyze peptides with the N-terminal amino acids glutamic acid (Glu) and aspartic acid (Asp). A possible application of PepA is the hydrolysis of Glu/Asp-rich food proteins such as wheat gluten and casein, increasing the flavor and solubility of food protein. In the present study, the gene encoding a Pep A from Lactococcus lactis ssp. lactis IL1403 was synthesized and introduced into Pichia pastoris GS115 (His4). Lc-Pep A was successfully expressed and secreted to the culture medium, followed by identification and purification to homogeneity. Characteristics study demonstrated that Lc-Pep A could specifically hydrolyze the substrates Glu-pNA and Asp-pNA with similar catalytic activity, and this was further confirmed by the kinetics parameters measured. Additionally, Lc-Pep A showed a broad thermostability and pH stability with an optimum temperature of 60 ℃ and an optimum pH of 8.0. The enzyme activity of Lc-Pep A was activated by metal ions Co2+, Mn2+, and Zn2+ but was strongly inhibited by Ni2+and Cu2+. The routine proteinase inhibitor had no effect on the activity of Lc-Pep A. However, Lc-Pep A was strongly inhibited by the metallopeptidase inhibitor, EDTA, and disulfide bond-reducing agents. The study may facilitate production and application of Lc-Pep A.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Biological Transport
/
Lactococcus lactis
/
Glutamic Acid
/
Culture Media
/
Glutamyl Aminopeptidase
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2023
Type:
Article
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