Overexpression in escherichia coli and characterization of the chloroplast triosephosphate isomerase from spinach
文献类型：期刊论文
| 作者 | Tang GL(唐功利) ; Wang YF(王燕芳) ; Bao JS(鲍建绍) ; Chen HB(陈海宝) |
| 刊名 | Protein Expr. Purif.
 |
| 出版日期 | 1999 |
| 卷号 | 16期号:3页码:432-439 |
| ISSN号 | 1046-5928 |
| 其他题名 | 菠菜叶绿体三糖磷酸酯异构化酶在大肠杆菌中高表达及其性能测定 |
| 通讯作者 | 陈海宝 |
| 英文摘要 | An important calvin cycle enzyme, chloroplast triosephosphate isomerase (cpTPI) from spinach, has been cloned and expressed in up to 15% of the total cell protein using the Pl expression vector in escherichia coli. An even higher level expression, up to 36% of the total protein, was achevd by replacing the nucleotide sequence between the ribosomal binding site and the initial codon, ATG, with an AT-rich sequence. Computer modeling revealed that the moderate change in the standard free energy (5'△G^o) of mRNA secondary structure in the translation initial region might be the major factor which led to the later high-level expression. The overexpressed spinach cpTPI was solubel and fully active and was abel to be purified beyond 95% purity by DEAE-Sepharose and sephadex G-75, and around 55 mg of purified enzymes was obtained from 1 liter of cultured bacteria. With D-glyceraldehyde 3-phosphate as substrate, Km(D-3-P) is 0.68 mM, V max (G-3-P) is 3.16×10^4 μmol/min.mg, and Kcat (G-3-P0 is 4.51×10^3/s; with dihydroxyacetone phosphate as substrate, the corresponding values are 7.27 mM, 1.04×10^3 μmol/min.mg, and 1.16 ×10^2/s, are 7.27 mM, 1.04×10^3 μmol/min.mg, and 1.16 ×10^2/s, respectively. |
| 学科主题 | 生命有机化学 |
| 收录类别 | SCI |
| 原文出处 | http://dx.doi.org/10.1006/prep.1999.1087 |
| 语种 | 英语 |
| WOS记录号 | WOS:000081814900009 |
| 公开日期 | 2013-06-19 |
| 源URL | [http://202.127.28.38/handle/331003/25741]  |
| 专题 | 上海有机化学研究所_生命有机化学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | Tang GL,Wang YF,Bao JS,et al. Overexpression in escherichia coli and characterization of the chloroplast triosephosphate isomerase from spinach[J]. Protein Expr. Purif.,1999,16(3):432-439. |
| APA | 唐功利,王燕芳,鲍建绍,&陈海宝.(1999).Overexpression in escherichia coli and characterization of the chloroplast triosephosphate isomerase from spinach.Protein Expr. Purif.,16(3),432-439. |
| MLA | 唐功利,et al."Overexpression in escherichia coli and characterization of the chloroplast triosephosphate isomerase from spinach".Protein Expr. Purif. 16.3(1999):432-439. |
入库方式： OAI收割
来源：上海有机化学研究所
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