Fusion expression of D-amino acid oxidase from Trignoposis variabilis with maltose binding protein and Vitreoscilla hemoglobin.
- Author:
Huimin YU
1
;
Xianfeng MA
;
Hui LUO
;
Cheng WEN
;
Zhongyao SHEN
Author Information
1. Department of Chemical Engineering, Tsinghua University, Beijing 100084, China. yuhm@tsinghua.edu.cn
- Publication Type:Journal Article
- MeSH:
Bacterial Proteins;
biosynthesis;
genetics;
Carrier Proteins;
biosynthesis;
genetics;
D-Amino-Acid Oxidase;
biosynthesis;
genetics;
Escherichia coli;
genetics;
metabolism;
Maltose-Binding Proteins;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Truncated Hemoglobins;
biosynthesis;
genetics;
Yeasts;
enzymology;
genetics
- From:
Chinese Journal of Biotechnology
2008;24(6):1004-1009
- CountryChina
- Language:Chinese
-
Abstract:
D-amino acid oxidase (DAAO) is one of important industrial enzymes. To increase the solubility and activity of the TvDAAO from Trignoposis variabilis expressed in recombinant Escherichia coli (E. coli), a maltose binding protein (MBP) and Vitreoscilla hemoglobin (VHb) was introduced to fuse with N-terminal of the TvDAAO, respectively. Fusion protein of MBP-TvDAAO was constitutively expressed in JM105/pMKC-DAAO and inductively expressed in JM105/pMKL-DAAO. With respect to the control strain of BL21 (DE3)/pET-DAAO without MBP fusion, the constitutive fusion expression obtained 28% of soluble protein with 3.7 folds of solubility improvement. As for the inductive fusion expression, corresponding results changed to 17% and 1.8 folds, respectively. However, the DAAO activity significantly decreased in the MBP-fusing expression. Fusion protein of VHb-TvDAAO was constructed and inductively expressed in BL21 (DE3)/pET-VDAAO. Its DAAO activity highly reached 3.24 u/mL in flask culture, about 90% increase in contrast to the control without VHb.
