Liu Changqing, Zheng Kai, Xu Ying, Stephen Lacmata Tamekou, Wang Jiming, Zhao Hongwei, Yue Tongqing, Nian Rui, Zhang Haibo, Xian Mo, Liu Huizhou
a CAS Key Laboratory of Biobased Materials , Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences , Qingdao , China ; University of Chinese Academy of Sciences , Beijing , China.
b Qilu University of Technology , Jinan , Shandong Province , P. R. China.
Prep Biochem Biotechnol. 2017 Sep 14;47(8):768-775. doi: 10.1080/10826068.2017.1342258. Epub 2017 Jun 23.
Soybean seed coat peroxidase (SBP) is a valuable enzyme having a broad variety of applications in analytical chemistry, biochemistry, and food processing. In the present study, the sscp gene (Gene ID: 548068) was optimized based on the preferred codon usage of Escherichia coli, synthesized, and expressed in E. coli BL21(DE3). SDS-PAGE and western blot analysis of this expressed protein revealed that its molecular weight is approximately 39 kDa. The effects of induction temperature, concentration of isopropyl-β-D-thiogalactoside and hemin, induction time, expression time were optimized to enhance SBP production with a maximum activity of 11.23 U/mL (8.64 U/mg total protein). Furthermore, the kinetics of enzyme-catalyzed reactions of recombinant protein was determined. When 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) was used as substrate, optimum reaction temperature and pH of the enzyme were 85°C and 5.0, respectively. The effects of metal ions on the enzymatic reaction were also further investigated. The SBP was successfully expressed in E. coli BL21(DE3) which would provide a more efficient production strategy for industrial applications of SBP.
