8JPW

Crystal Structure of Single-chain L-Glutamate Oxidase Mutant from Streptomyces sp. X-119-6

Summary for 8JPW

• Entry DOI: 10.2210/pdb8jpw/pdb
• Descriptor: L-glutamate oxidase, FLAVIN-ADENINE DINUCLEOTIDE, 2-OXOGLUTARIC ACID, ... (5 entities in total)
• Functional Keywords: amino acid oxidase, mutant, oxidoreductase
• Biological source: Streptomyces sp. X-119-6
• Total number of polymer chains: 1
• Total formula weight: 74369.08

Authors

Yamaguchi, H.,Takahashi, K.,Tatsumi, M.,Tagami, U.,Mizukoshi, T.,Miyano, H.,Sugiki, M. (deposition date: 2023-06-13, release date: 2023-08-09, Last modification date: 2024-05-08)

Primary citation

Yamaguchi, H.,Takahashi, K.,Tatsumi, M.,Tagami, U.,Mizukoshi, T.,Miyano, H.,Sugiki, M.
Development of a novel single-chain l-glutamate oxidase from Streptomyces sp. X-119-6 by inserting flexible linkers.
Enzyme.Microb.Technol., 170:110287-110287, 2023

PubMed Abstract: L-glutamate oxidase (LGOX, EC: 1.4.3.11) is an oxidoreductase that catalyzes L-glutamate deamination. LGOX from Streptomyces sp. X-119-6 is used widely for L-glutamate quantification in research and industrial applications. This enzyme encoded as a single precursor chain that undergoes post-translational cleavage to four fragments by an endogenous protease to become highly active. Efficient preparation of active LGOX by heterologous expression without proteolysis process should be indispensable for wide application of this enzyme. Thus, developing an LGOX that requires no protease treatment should expand the potential applications of recombinant LGOX. In this report, we succeeded in obtaining an active single-chain LGOX by connecting the four fragments of the mature form with insertion of flexible linkers. The most active single-chain mutant showed the similar activity to that of the mature form from Streptomyces sp. X-119-6. The structure of this mutant was determined at 2.9 Å resolution by X-ray crystallography. It was revealed that this single-stranded mutant had the similar conformation to that of mature form. This single-chain LGOX can be produced efficiently and should expand LGOX applications.

PubMed: 37487431
DOI: 10.1016/j.enzmictec.2023.110287

Experimental method

X-RAY DIFFRACTION (2.66 Å)