Expression of the glyceraldehyde‐3‐phosphate dehydrogenase gene from the extremely thermophilic archaebacterium Methanothermus fervidus in E. coli

FEBS Letters

Hensel, Reinhard¹  Lehmacher, Anselm¹  Bode, Wolfram¹  Fabry, Stefan¹ 

[1] Max-Planck-Institut für Biochemie, Am Klopferspitz, D-8033 Martinsried, FRG

关键词: Methanobacteria;    Enzyme expression;    Protein thermostability;    Protein crystallography;   

DOI  :  10.1016/0014-5793(88)80204-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The gene of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the extremely thermophilic archaebacterium Methanothermus fervidus (growth optimum 82°C) was cloned in vector pJF118EH and expressed in E. coli cells. As shown by molecular mass determination, protein sequencing, heat stability, and substrate saturation kinetics, the enzyme synthesized in E. coli is identical to the original enzyme from M. fervidus. The high thermostability of the E. coli-produced M. fervidus GAPDH allows rapid purification to homogeneity. From this enzyme protein crystals were grown which proved to be suitable for X-ray analysis. The crystals are of tetragonal space group P4₁22 and contain a dimer per asymmetric unit.

【 授权许可】

Unknown