Purification and characterization of Archaeoglobus fulgidus shikimate 5-dehydrogenase

Author

Lim, Sierin ; Kim, Chihee ; Schröder, Imke ; Monbouquette, Harold G.

Author_Institution

Biomed. Eng. Interdepartmental Program, California Univ., Los Angeles, CA, USA

Volume

1

fYear

2002

fDate

2002

Firstpage

701

Abstract

Shikimate 5-dehydrogenase (EC 1.1.1.25) is an important enzyme of the aromatic amino acid biosynthesis pathway. The shikimate 5-dehydrogenase (SDH) gene from the hyperthermophile Archaeoglobus fulgidus was PCR cloned and over-expressed in E. coli. The resulting recombinant enzyme with a M_r of 27,000 was purified to homogeneity. The enzyme had a specific activity of 727 U/mg at 87°C, and exhibited K_ms for shikimate and NADP⁺ of 0.17 ± 0.03 mM and 0.19 ± 0.01, respectively. At 87°C, the half life of the SDH was 2 hours. At 60°C and a specific activity of 104 U/mg, the half life was 17 days. The combination of high stability and activity for this archaeal SDH may make it useful for industrial chiral synthesis.

Keywords

biochemistry; biotechnology; biothermics; genetics; microorganisms; proteins; 17 day; 2 hr; 60 C; 87 C; Archaeoglobus fulgidus shikimate 5-dehydrogenase; E. coli; NADP⁺; aromatic amino acid biosynthesis pathway; enzyme characterization; enzyme purification; half life; hyperthermophile; industrial chiral synthesis; Amino acids; Archaea; Biochemistry; Electrons; Microorganisms; Organisms; Proteins; Purification; Synchronous digital hierarchy; Temperature;

fLanguage

English

Publisher

ieee

Conference_Titel

Engineering in Medicine and Biology, 2002. 24th Annual Conference and the Annual Fall Meeting of the Biomedical Engineering Society EMBS/BMES Conference, 2002. Proceedings of the Second Joint

ISSN

1094-687X

Print_ISBN

0-7803-7612-9

Type

conf

DOI

10.1109/IEMBS.2002.1137025

Filename

1137025