Purification and Characterization of Ethyl 2-Methyl-3-oxobutanoate Reductase fromKlebsiella pneumoniaeIF03319
1
a Integrated Technology Laboratories , Takeda Chemical Industries Ltd. , Wadai 10, Tsukuba, Ibaraki 300-42 , Japan.
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Publication type: Journal Article
Publication date: 1996-01-01
scimago Q3
wos Q3
SJR: 0.405
CiteScore: 3.1
Impact factor: 1.3
ISSN: 09168451, 13476947
PubMed ID:
27281138
Organic Chemistry
Biochemistry
Molecular Biology
General Medicine
Analytical Chemistry
Applied Microbiology and Biotechnology
Biotechnology
Abstract
An enzyme that catalyzes a reduction of ethyl 2-methyl-3-oxobutanoate (1) to ethyl (2R,3S) 3-hydroxy- 2-methylbutanoate was found in Klebsiella pneumoniae IFO 3319 cells. The enzyme was isolated from the cells and purified 250-fold by ammonium sulfate fractionation, ion exchange chromatography, affinity chromatography, and gel filtration. The purified enzyme was found to be a monomer protein with a molecular weight of approximately 31,000 and an isoelectric point of 6.2. It was NADPH-dependent and had maximum activity at pH 7.0 and 45°C for the reduction and at pH 10.0 and 45°C for oxidation. The Km's at pH 7.0 were 5.6 mM for 1 and 12.5 mM for benzyl 2-methyl-3-oxobutanoate, respectively. Esters of 2-oxocycloalkane carboxylic acids as well as esters of 2-methyl-3-oxobutanoic acid served as substrates, and the corresponding reduced products were obtained with high stereoselectivity.
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Miya H., KAWADA M., Sugiyama Y. Purification and Characterization of Ethyl 2-Methyl-3-oxobutanoate Reductase fromKlebsiella pneumoniaeIF03319 // Bioscience, Biotechnology and Biochemistry. 1996. Vol. 60. No. 5. pp. 760-764.
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Miya H., KAWADA M., Sugiyama Y. Purification and Characterization of Ethyl 2-Methyl-3-oxobutanoate Reductase fromKlebsiella pneumoniaeIF03319 // Bioscience, Biotechnology and Biochemistry. 1996. Vol. 60. No. 5. pp. 760-764.
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RIS
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TY - JOUR
DO - 10.1271/bbb.60.760
UR - https://doi.org/10.1271/bbb.60.760
TI - Purification and Characterization of Ethyl 2-Methyl-3-oxobutanoate Reductase fromKlebsiella pneumoniaeIF03319
T2 - Bioscience, Biotechnology and Biochemistry
AU - Miya, Hiroyuki
AU - KAWADA, Mitsuru
AU - Sugiyama, Yoshio
PY - 1996
DA - 1996/01/01
PB - Taylor & Francis
SP - 760-764
IS - 5
VL - 60
PMID - 27281138
SN - 0916-8451
SN - 1347-6947
ER -
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@article{1996_Miya,
author = {Hiroyuki Miya and Mitsuru KAWADA and Yoshio Sugiyama},
title = {Purification and Characterization of Ethyl 2-Methyl-3-oxobutanoate Reductase fromKlebsiella pneumoniaeIF03319},
journal = {Bioscience, Biotechnology and Biochemistry},
year = {1996},
volume = {60},
publisher = {Taylor & Francis},
month = {jan},
url = {https://doi.org/10.1271/bbb.60.760},
number = {5},
pages = {760--764},
doi = {10.1271/bbb.60.760}
}
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MLA
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Miya, Hiroyuki, et al. “Purification and Characterization of Ethyl 2-Methyl-3-oxobutanoate Reductase fromKlebsiella pneumoniaeIF03319.” Bioscience, Biotechnology and Biochemistry, vol. 60, no. 5, Jan. 1996, pp. 760-764. https://doi.org/10.1271/bbb.60.760.