Characterization of a D-mannitol oxidase from Paenibacillus sp. and its application in the preparation of D-mannose.
- Author:
Ran LI
1
;
Cong SONG
1
;
Xiang ZHANG
1
;
Zhenhua JIA
1
Author Information
- Publication Type:Journal Article
- Keywords: D-mannitol oxidase; D-mannose; enzyme property; whole cell catalysis
- MeSH: Recombinant Proteins/metabolism*; Paenibacillus/metabolism*; Mannose/metabolism*; Escherichia coli/metabolism*; Mannitol/metabolism*
- From: Chinese Journal of Biotechnology 2023;39(11):4682-4693
- CountryChina
- Language:Chinese
- Abstract: D-mannose has many functional activities and is widely used in food, medicine, agriculture and other industries. D-mannitol oxidase that can efficiently convert D-mannitol into D-mannose has potential application in the enzymatic preparation of D-mannose. A D-mannitol oxidase (PsOX) was found from Paenibacillus sp. HGF5. The similarity between PsOX and the D-mannitol oxidase (AldO) from Streptomyces coelicolor was 50.94%. The molecular weight of PsOX was about 47.4 kDa. A recombinant expression plasmid pET-28a-PsOX was constructed and expressed in Escherichia coli BL21(DE3). The Km and kcat/Km values of PsOX for D-mannitol were 5.6 mmol/L and 0.68 L/(s·mmol). Further characterization of PsOX showed its optimal pH and temperature were 7.0 and 35 ℃, respectively, while its enzyme activity could be stably remained below 60 ℃. The molar conversion rate of 400 mmol/L D-mannitol by PsOX was 95.2%. The whole cells of PsOX and AldO were used to catalyze 73 g/L D-mannitol respectively. The reaction catalyzed by PsOX completed in 9 h and 70 g/L D-mannose was produced. PsOX showed a higher catalytic efficiency compared to that of AldO. PsOX may facilitate the enzymatic preparation of D-mannose as a novel D-mannose oxidase.
