Heterologous expression and characterization of Aspergillus oryzae acidic protease in Pichia pastoris.
- Author:
Xiaoping YUE
1
;
Peng CHEN
2
;
Yueming ZHU
2
;
Yan ZENG
2
;
Hanmin LIU
3
;
Hongyan LIU
3
;
Min WANG
1
;
Yuanxia SUN
2
Author Information
- Publication Type:Journal Article
- Keywords: Aspergillums oryzae; Pichia pastoris; acidic protease enzyme; enzymatic properties; heterologous expression
- MeSH: Aspergillus oryzae; Cloning, Molecular; Endopeptidases; Hydrogen-Ion Concentration; Pichia; Recombinant Proteins; Temperature
- From: Chinese Journal of Biotechnology 2019;35(3):415-424
- CountryChina
- Language:Chinese
- Abstract: Acid protease, an important aspartic protease, has been widely used in food, pharmaceutical and tanning industries. To promote the research and application of acid protease, an acid protease gene (pepA) from Aspergillus oryzae was obtained from fermented soy based on metagenome sequencing, and then cloned and transformed into Pichia pastoris GS115 for heterologous expression. The characteristic of recombinant PepA was also investigated. The activity of acid protease in the culture supernatant of P. pastoris was 50.62 U/mL. The molecular mass of PepA was about 50 kDa, and almost no other proteins in the supernatant were observed, as shown by SDS-PAGE. The optimum pH and temperature of PepA were determined as pH 4.5 and 50 ℃. Mn²⁺ and Cu²⁺ enhanced the activity of PepA, whereas Fe³⁺, Fe²⁺ and Ca² had inhibitory effects on its activity. The above findings can provide guidance for heterologous expression and industrial application of acid protease from Aspergillus oryzae.
