Characterization of native glutamate dehydrogenase from an aerobic hyperthermophilic archaeon Aeropyrum pernix K1

• Published in: Applied microbiology and biotechnology Vol. 56; no. 3-4; pp. 388 - 394
• Main Authors: HELIANTI, I, MORITA, Y, YAMAMURA, A, MURAKAMI, Y, YOKOYAMA, K, TAMIYA, E
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01-08-2001; Springer Nature B.V
• Subjects: Acetonitrile; Aerobiosis; Aeropyrum pernix; Amino Acid Sequence; Amino acids; Archaea; Bacteria; Bacteriology; Base Sequence; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Dehydrogenase; Dehydrogenases; Desulfurococcaceae - enzymology; Desulfurococcaceae - growth & development; Enzyme Stability; Enzymes; Ethanol; Fundamental and applied biological sciences. Psychology; Glutamate Dehydrogenase - chemistry; Glutamate Dehydrogenase - isolation & purification; Glutamate Dehydrogenase - metabolism; Kinetics; methionine; Microbiology; Miscellaneous; Mission oriented research; Molecular Sequence Data; Organic solvents; Solvents - pharmacology; Substrate Specificity; Temperature; Glutamate dehydrogenase; Aerobiosis; Purification; Enzymatic activity; Archaeobacteria; Enzyme; Hyperthermophily; Bacteria; Oxidoreductases; Kinetic parameter; Physicochemical properties; Thermal stability
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s002530100575

Glutamate dehydrogenase (GDH) was purified and characterized from an aerobic hyperthermophilic archaeon Aeropyrum pernix (A. pernix) K1. The enzyme has a hexameric structure with a native molecular mass of about 285 +/- 15 kDa. It was specific for NADP and thermostable (74% activity was remained after 5 h incubation at 100 degrees C). The activity of the enzyme increased in the presence of polar water-miscible organic solvents such as acetonitrile, methanol, and ethanol. The N-terminal sequence of GDH is Met-Gln-Pro-Thr-Asp-Pro-Leu-Glu-Glu-Ala. This sequence, except for the methionine, corresponds to amino acids 7-15 of the open reading frame (ORF) encoding the predicted GDH (ORF APE 1386). In the ORF nucleotide sequence, the codon TTG appears at the position of the methionine, suggesting that the leucine codon might be recognized as an initiation codon and translated to methionine in A. pernix GDH.