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Alteration of coenzyme specificity of malate dehydrogenase from Streptomyces coelicolor A3(2) by site-directed mutagenesis.
Genet Mol Res. 2014 Jul 29; 13(3):5758-66.GM

Abstract

We describe here for the first time the alteration of coenzyme specificity of malate dehydrogenase (MDH) from Streptomyces coelicolor A3(2) (ScMDH). In the present study, we replaced four amino acid residues in the Rossmann fold (βB-αC) region of NADH-dependent ScMDH by site-directed mutagenesis with those of NADPH-dependent MDH (Glu42Gly, Ile43Ser, Pro45Arg, and Ala46Ser). The coenzyme specificity of the mutant enzyme (ScMDH-T4) was examined. Coenzyme specificity of ScMDH-T4 was shifted 2231.3-fold toward NADPH using kcat/Km(coenzyme) as the measurement of coenzyme specificity. Accordingly, the effect of the replacements on coenzyme specificity is discussed. Our work provides further insight into the coenzyme specificity of ScMDH.

Authors+Show Affiliations

Institute of Molecular Biology and Biotechnology, Key Laboratory of Molecular Evolution and Biodiversity, Key Laboratory of the Biotic Environment and Ecological Safety in Anhui Province, Anhui Normal University, Wuhu, Anhui, China.Institute of Molecular Biology and Biotechnology, Key Laboratory of Molecular Evolution and Biodiversity, Key Laboratory of the Biotic Environment and Ecological Safety in Anhui Province, Anhui Normal University, Wuhu, Anhui, China.Institute of Molecular Biology and Biotechnology, Key Laboratory of Molecular Evolution and Biodiversity, Key Laboratory of the Biotic Environment and Ecological Safety in Anhui Province, Anhui Normal University, Wuhu, Anhui, China.Institute of Molecular Biology and Biotechnology, Key Laboratory of Molecular Evolution and Biodiversity, Key Laboratory of the Biotic Environment and Ecological Safety in Anhui Province, Anhui Normal University, Wuhu, Anhui, China.Institute of Molecular Biology and Biotechnology, Key Laboratory of Molecular Evolution and Biodiversity, Key Laboratory of the Biotic Environment and Ecological Safety in Anhui Province, Anhui Normal University, Wuhu, Anhui, China gpz1996@yahoo.com.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25117334

Citation

Ge, Y D., et al. "Alteration of Coenzyme Specificity of Malate Dehydrogenase From Streptomyces Coelicolor A3(2) By Site-directed Mutagenesis." Genetics and Molecular Research : GMR, vol. 13, no. 3, 2014, pp. 5758-66.
Ge YD, Song P, Cao ZY, et al. Alteration of coenzyme specificity of malate dehydrogenase from Streptomyces coelicolor A3(2) by site-directed mutagenesis. Genet Mol Res. 2014;13(3):5758-66.
Ge, Y. D., Song, P., Cao, Z. Y., Wang, P., & Zhu, G. P. (2014). Alteration of coenzyme specificity of malate dehydrogenase from Streptomyces coelicolor A3(2) by site-directed mutagenesis. Genetics and Molecular Research : GMR, 13(3), 5758-66. https://doi.org/10.4238/2014.July.29.3
Ge YD, et al. Alteration of Coenzyme Specificity of Malate Dehydrogenase From Streptomyces Coelicolor A3(2) By Site-directed Mutagenesis. Genet Mol Res. 2014 Jul 29;13(3):5758-66. PubMed PMID: 25117334.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Alteration of coenzyme specificity of malate dehydrogenase from Streptomyces coelicolor A3(2) by site-directed mutagenesis. AU - Ge,Y D, AU - Song,P, AU - Cao,Z Y, AU - Wang,P, AU - Zhu,G P, Y1 - 2014/07/29/ PY - 2014/8/14/entrez PY - 2014/8/15/pubmed PY - 2015/4/24/medline SP - 5758 EP - 66 JF - Genetics and molecular research : GMR JO - Genet. Mol. Res. VL - 13 IS - 3 N2 - We describe here for the first time the alteration of coenzyme specificity of malate dehydrogenase (MDH) from Streptomyces coelicolor A3(2) (ScMDH). In the present study, we replaced four amino acid residues in the Rossmann fold (βB-αC) region of NADH-dependent ScMDH by site-directed mutagenesis with those of NADPH-dependent MDH (Glu42Gly, Ile43Ser, Pro45Arg, and Ala46Ser). The coenzyme specificity of the mutant enzyme (ScMDH-T4) was examined. Coenzyme specificity of ScMDH-T4 was shifted 2231.3-fold toward NADPH using kcat/Km(coenzyme) as the measurement of coenzyme specificity. Accordingly, the effect of the replacements on coenzyme specificity is discussed. Our work provides further insight into the coenzyme specificity of ScMDH. SN - 1676-5680 UR - https://www.unboundmedicine.com/medline/citation/25117334/Alteration_of_coenzyme_specificity_of_malate_dehydrogenase_from_Streptomyces_coelicolor_A3_2__by_site_directed_mutagenesis_ L2 - http://www.geneticsmr.com/articles/3393 DB - PRIME DP - Unbound Medicine ER -