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Crystal Structure and Biochemical Characterization of Ketol-Acid Reductoisomerase from Corynebacterium glutamicum.
J Agric Food Chem. 2019 Aug 07; 67(31):8527-8535.JA

Abstract

l-Valine belongs to the branched-chain amino acids (BCAAs) and is an essential amino acid that is crucial for all living organisms. l-Valine is industrially produced by the nonpathogenic bacterium Corynebacterium glutamicum and is synthesized by the BCAA biosynthetic pathway. Ketol-acid reductoisomerase (KARI) is the second enzyme in the BCAA pathway and catalyzes the conversion of (S)-2-acetolactate into (R)-2,3-dihydroxy-isovalerate, or the conversion of (S)-2-aceto-2-hydroxybutyrate into (R)-2,3-dihydroxy-3-methylvalerate. To elucidate the enzymatic properties of KARI from C. glutamicum (CgKARI), we successfully produced CgKARI protein and determined its crystal structure in complex with NADP+ and two Mg2+ ions. Based on the complex structure, docking simulations, and site-directed mutagenesis experiments, we revealed that CgKARI belongs to Class I KARI and identified key residues involved in stabilization of the substrate, metal ions, and cofactor. Furthermore, we confirmed the difference in the binding of metal ions that depended on the conformational change.

Authors+Show Affiliations

School of Life Sciences, BK21 Plus KNU Creative BioResearch Group , Kyungpook National University , Daehak-ro 80, Buk-ku , Daegu 702-701 , Korea. KNU Institute for Microorganisms , Kyungpook National University , Daegu 41566 , Republic of Korea.School of Life Sciences, BK21 Plus KNU Creative BioResearch Group , Kyungpook National University , Daehak-ro 80, Buk-ku , Daegu 702-701 , Korea. KNU Institute for Microorganisms , Kyungpook National University , Daegu 41566 , Republic of Korea.School of Life Sciences, BK21 Plus KNU Creative BioResearch Group , Kyungpook National University , Daehak-ro 80, Buk-ku , Daegu 702-701 , Korea. KNU Institute for Microorganisms , Kyungpook National University , Daegu 41566 , Republic of Korea.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31298526

Citation

Lee, Donghoon, et al. "Crystal Structure and Biochemical Characterization of Ketol-Acid Reductoisomerase From Corynebacterium Glutamicum." Journal of Agricultural and Food Chemistry, vol. 67, no. 31, 2019, pp. 8527-8535.
Lee D, Hong J, Kim KJ. Crystal Structure and Biochemical Characterization of Ketol-Acid Reductoisomerase from Corynebacterium glutamicum. J Agric Food Chem. 2019;67(31):8527-8535.
Lee, D., Hong, J., & Kim, K. J. (2019). Crystal Structure and Biochemical Characterization of Ketol-Acid Reductoisomerase from Corynebacterium glutamicum. Journal of Agricultural and Food Chemistry, 67(31), 8527-8535. https://doi.org/10.1021/acs.jafc.9b03262
Lee D, Hong J, Kim KJ. Crystal Structure and Biochemical Characterization of Ketol-Acid Reductoisomerase From Corynebacterium Glutamicum. J Agric Food Chem. 2019 Aug 7;67(31):8527-8535. PubMed PMID: 31298526.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Crystal Structure and Biochemical Characterization of Ketol-Acid Reductoisomerase from Corynebacterium glutamicum. AU - Lee,Donghoon, AU - Hong,Jiyeon, AU - Kim,Kyung-Jin, Y1 - 2019/07/25/ PY - 2019/7/13/pubmed PY - 2019/8/17/medline PY - 2019/7/13/entrez KW - KW - -valine KW - ketol-acid reductoisomerase SP - 8527 EP - 8535 JF - Journal of agricultural and food chemistry JO - J. Agric. Food Chem. VL - 67 IS - 31 N2 - l-Valine belongs to the branched-chain amino acids (BCAAs) and is an essential amino acid that is crucial for all living organisms. l-Valine is industrially produced by the nonpathogenic bacterium Corynebacterium glutamicum and is synthesized by the BCAA biosynthetic pathway. Ketol-acid reductoisomerase (KARI) is the second enzyme in the BCAA pathway and catalyzes the conversion of (S)-2-acetolactate into (R)-2,3-dihydroxy-isovalerate, or the conversion of (S)-2-aceto-2-hydroxybutyrate into (R)-2,3-dihydroxy-3-methylvalerate. To elucidate the enzymatic properties of KARI from C. glutamicum (CgKARI), we successfully produced CgKARI protein and determined its crystal structure in complex with NADP+ and two Mg2+ ions. Based on the complex structure, docking simulations, and site-directed mutagenesis experiments, we revealed that CgKARI belongs to Class I KARI and identified key residues involved in stabilization of the substrate, metal ions, and cofactor. Furthermore, we confirmed the difference in the binding of metal ions that depended on the conformational change. SN - 1520-5118 UR - https://www.unboundmedicine.com/medline/citation/31298526/Crystal_Structure_and_Biochemical_Characterization_of_Ketol-acid_Reductoisomerase_from_Corynebacterium_glutamicum L2 - https://dx.doi.org/10.1021/acs.jafc.9b03262 DB - PRIME DP - Unbound Medicine ER -