Expression and characterization of soybean seed coat peroxidase in Escherichia coli BL21(DE3)

doi:10.1080/10826068.2017.1342258

QIBEBT-IR > 生物基材料组群

	Expression and characterization of soybean seed coat peroxidase in Escherichia coli BL21(DE3)
	Liu, Changqing 1,2; Zheng, Kai 3; Xu, Ying 1,2; Stephen, Lacmata Tamekou 1,2,5; Wang, Jiming 1,2; Zhao, Hongwei 1,2; Yue, Tongqing 4; Nian, Rui 1,2; Zhang, Haibo 1,2; Xian, Mo 1,2; Liu, Huizhou 1,2
	2017
发表期刊	PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
卷号	47 期号:8 页码:768-775
摘要	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 39kDa. The effects of induction temperature, concentration of isopropyl--D-thiogalactoside and hemin, induction time, expression time were optimized to enhance SBP production with a maximum activity of 11.23U/mL (8.64U/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 degrees 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.
文章类型	Article
关键词	Abts Escherichia Coli Heterologous Expression Ph Recombinant Protein Soybean Seed Coat Peroxidase Temperature
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
DOI	10.1080/10826068.2017.1342258
关键词[WOS]	HORSERADISH-PEROXIDASE ; ANIONIC PEROXIDASE ; MOLECULAR-CLONING ; HEME ; TISSUE ; PURIFICATION ; STABILITY ; CULTURE ; TOBACCO ; SYSTEM
收录类别	SCI
语种	英语
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
项目资助者	National Natural Science Foundation of China (NSF)(31400084) ; Natural Science Foundation of Shandong Province(ZR2014CQ008) ; Taishan Scholars Climbing Program of Shandong(TSPD20150210) ; Youth Innovation Promotion Association CAS(2017252)
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS记录号	WOS:000410881700004
引用统计	被引频次：7[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/9726
专题	生物基材料组群
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biobased Mat, 189 Songling Rd, Qingdao 266101, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.Qilu Univ Technol, Jinan, Shandong, Peoples R China 4.Qingdao Univ, Coll Life Sci, Qingdao, Peoples R China 5.Univ Dschang, Dept Biochem, Lab Microbiol & Antimicrobials Subst, Fac Sci, Dschang, Cameroon
推荐引用方式 GB/T 7714	Liu, Changqing,Zheng, Kai,Xu, Ying,et al. Expression and characterization of soybean seed coat peroxidase in Escherichia coli BL21(DE3)[J]. PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY,2017,47(8):768-775.
APA	Liu, Changqing.,Zheng, Kai.,Xu, Ying.,Stephen, Lacmata Tamekou.,Wang, Jiming.,...&Liu, Huizhou.(2017).Expression and characterization of soybean seed coat peroxidase in Escherichia coli BL21(DE3).PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY,47(8),768-775.
MLA	Liu, Changqing,et al."Expression and characterization of soybean seed coat peroxidase in Escherichia coli BL21(DE3)".PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY 47.8(2017):768-775.