Expression and characterization of soybean seed coat peroxidase in Escherichia coli BL21(DE3)
文献类型：期刊论文
| 作者 | Liu, Changqing1,2; Zheng, Kai3; Xu, Ying1,2; Stephen, Lacmata Tamekou1,2,5; Wang, Jiming1,2; Zhao, Hongwei1,2; Yue, Tongqing4; Nian, Rui1,2; Zhang, Haibo1,2; Xian, Mo1,2 |
| 刊名 | PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
 |
| 出版日期 | 2017 |
| 卷号 | 47期号:8页码:768-775 |
| 关键词 | Abts Escherichia Coli Heterologous Expression Ph Recombinant Protein Soybean Seed Coat Peroxidase Temperature |
| DOI | 10.1080/10826068.2017.1342258 |
| 文献子类 | Article |
| 英文摘要 | 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. |
| WOS关键词 | HORSERADISH-PEROXIDASE ; ANIONIC PEROXIDASE ; MOLECULAR-CLONING ; HEME ; TISSUE ; PURIFICATION ; STABILITY ; CULTURE ; TOBACCO ; SYSTEM |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000410881700004 |
| 资助机构 | 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) |
| 源URL | [http://ir.qibebt.ac.cn/handle/337004/9726]  |
| 专题 | 青岛生物能源与过程研究所_材料生物技术研究中心 |
| 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. |
入库方式： OAI收割
来源：青岛生物能源与过程研究所
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