1. Characterization of the linoleic acid Δ9 hydratase catalyzing the first step of polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a.
- Author
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Takeuchi M, Kishino S, Hirata A, Park SB, Kitamura N, and Ogawa J
- Subjects
- Flavin-Adenine Dinucleotide metabolism, Hydrogen-Ion Concentration, Hydrolases metabolism, Kinetics, Lipid Metabolism, NAD metabolism, Oleic Acids chemistry, Oleic Acids metabolism, Substrate Specificity, Temperature, Biocatalysis, Lactobacillus plantarum enzymology, Lactobacillus plantarum metabolism, Linoleic Acid metabolism
- Abstract
Linoleic acid Δ9 hydratase, which is involved in linoleic acid saturation metabolism of Lactobacillus plantarum AKU 1009a, was cloned, expressed as a his-tagged recombinant enzyme, purified with an affinity column, and characterized. The enzyme required FAD as a cofactor and its activity was enhanced by NADH. The maximal activities for the hydration of linoleic acid and for the dehydration of 10-hydroxy-cis-12-octadecenoic acid (HYA) were observed at 37 °C in buffer at pH 5.5 containing 0.5 M NaCl. Free C16 and C18 fatty acids with cis-9 double bonds and 10-hydroxy fatty acids served as substrates for the hydration and dehydration reactions, respectively. The apparent Km value for linoleic acid was estimated to be 92 μM, with a kcat of 2.6∙10(-2) s(-1) and a Hill factor of 3.3. The apparent Km value for HYA was estimated to be 98 μM, with a kcat of 1.2∙10(-3) s(-1)., (Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)
- Published
- 2015
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