Phosphoserine aminotransferase from Bacillus circulans subsp alkalophilus: Purification, gene cloning and sequencing

: Battchikova N, Himanen JP, Ahjolahti M, Korpela T

Publisher: ELSEVIER SCIENCE BV

: 1996

: Biochimica et Biophysica Acta: Protein Structure and Molecular Enzymology

: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY

: BBA-PROTEIN STRUCT M

: 1295

: 2

: 187

: 194

: 8

: 0167-4838

DOI: https://doi.org/10.1016/0167-4838(96)00039-8

Two peaks of aspartate aminotransferase (AspAT) catalytic activity were observed during DEAE chromatography of a protein extract from alkalophilic B. circulans. The enzyme purified from the major peak appeared to be not aspartate but phosphoserine aminotransferase (PSAT) with a considerably high AspAT side activity. The sequence of the enzyme N-terminus was determined, and the PSAT gene was cloned as two separate fragments. DNA sequencing revealed the open reading frame for the PSAT starting from TTG, putative ribosomal binding site and terminator of transcription. The PSAT gene encodes a protein of 361 amino acids (M(r) 39 793) which shows moderate homology to other known phosphoserine aminotransferases (36-46% of identity, 60-64% of similarity). The PSAT from the alkalophile shares with all of them the consensus sequence pattern around the pyridoxal S-phosphate attachment site.