Back to Search
Start Over
Structure and activity of NADPH-dependent reductase Q1EQE0 from Streptomyces kanamyceticus, which catalyses the R-selective reduction of an imine substrate
- Source :
- ChemBioChem
- Publication Year :
- 2013
-
Abstract
- NADPH-dependent oxidoreductase Q1EQE0 from Streptomyces kanamyceticus catalyzes the asymmetric reduction of the prochiral monocyclic imine 2-methyl-1-pyrroline to the chiral amine (R)-2-methylpyrrolidine with >99% ee, and is thus of interest as a potential biocatalyst for the production of optically active amines. The structures of Q1EQE0 in native form, and in complex with the nicotinamide cofactor NADPH have been solved and refined to a resolution of 2.7 A. Q1EQE0 functions as a dimer in which the monomer consists of an N-terminal Rossman-fold motif attached to a helical C-terminal domain through a helix of 28 amino acids. The dimer is formed through reciprocal domain sharing in which the C-terminal domains are swapped, with a substrate-binding cleft formed between the N-terminal subunit of monomer A and the C-terminal subunit of monomer B. The structure is related to those of known β-hydroxyacid dehydrogenases, except that the essential lysine, which serves as an acid/base in the (de)protonation of the nascent alcohol in those enzymes, is replaced by an aspartate residue, Asp187 in Q1EQE0. Mutation of Asp187 to either asparagine or alanine resulted in an inactive enzyme.
- Subjects :
- Stereochemistry
Dimer
Imine
Reductase
010402 general chemistry
Crystallography, X-Ray
01 natural sciences
Biochemistry
Substrate Specificity
chemistry.chemical_compound
Bacterial Proteins
Oxidoreductase
Catalytic Domain
Molecular Biology
Alanine
chemistry.chemical_classification
Binding Sites
biology
010405 organic chemistry
Chemistry
Organic Chemistry
biology.organism_classification
Recombinant Proteins
Streptomyces
0104 chemical sciences
Amino acid
Protein Structure, Tertiary
Kinetics
Molecular Medicine
Imines
Oxidoreductases
Dimerization
Alpha helix
NADP
Streptomyces kanamyceticus
Subjects
Details
- ISSN :
- 14397633
- Volume :
- 14
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- Chembiochem : a European journal of chemical biology
- Accession number :
- edsair.doi.dedup.....ee57991a1c1c702f1a4618705c0fe7f0