The glyoxysomal 3‐ketoacyl‐CoA thiolase precursor from Brassica napus has enzymatic activity when synthesized in Escherichia coli

FEBS Letters

Brandt, Anders²  Svendsen, Ib¹  Thomsen, Karl Kristian²  Olesen, Christian² 

[1] Department of Chemistry, Carlsberg Laboratory, Gamle Carlsbergvej 10, DK-2500 Copenhagen Valby, Denmark;Department of Physiology, Carlsberg Laboratory, Gamle Carlsbergvej 10, DK-2500 Copenhagen Valby, Denmark

关键词: Expression;    Microbody;    Presequence;   

DOI  :  10.1016/S0014-5793(97)00766-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Glyoxysomal 3-ketoacyl-CoA thiolase is the last enzyme in the β-oxidation of fatty acids in plant glyoxysomes. A full-length cDNA of the glyoxysomal 3-ketoacyl-CoA thiolase from Brassica napus and a truncated version, lacking the N-terminal targeting signal were cloned in a T7 promoter-based vector. Both recombinant proteins were expressed in Escherichia coli and activity was measured. Full-length and truncated 3-ketoacyl-CoA thiolase have comparable activity in E. coli. Moreover, full-length 3-ketoacyl-CoA thiolase was purified from E. coli and N-terminal sequencing of the protein confirmed that the precursor form indeed is enzymatically active.

【 授权许可】

Unknown