Premium
Expression of the glyceraldehyde‐3‐phosphate dehydrogenase gene from the extremely thermophilic archaebacterium Methanothermus fervidus in E. coli
Author(s) -
Fabry Stefan,
Lehmacher Anselm,
Bode Wolfram,
Hensel Reinhard
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80204-7
Subject(s) - thermostability , glyceraldehyde 3 phosphate dehydrogenase , thermophile , dehydrogenase , biochemistry , enzyme , biology , glyceraldehyde , chemistry , microbiology and biotechnology
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 P4 1 22 and contain a dimer per asymmetric unit.