期刊
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
卷 119, 期 6, 页码 636-641出版社
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2014.10.022
关键词
Lactic acid bacteria; Hydroxy fatty acid; Hydratase; Hydration; Dehydration
资金
- Industrial Technology Research Grant Program [07A08005a]
- New Energy and Industrial Technology Development Organization (NEDO) of Japan
- Centers of Excellence for Microbial-Process Development Pioneering Future Production Systems from the Ministry of Education, Culture, Sports, Science, and Technology of Japan
- Bio-Oriented Technology Research Advancement Institution of Japan
- Advanced Low Carbon Technology Research and Development Program of Japan
- Japan Society for the Promotion of Science for Young Scientists
- [19780056]
- [16688004]
- [18208009]
- Grants-in-Aid for Scientific Research [14J00686] Funding Source: KAKEN
Linoleic acid Delta 9 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 degrees 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 K-m value for linoleic acid was estimated to be 92 mu M, with a k(cat) of 2.6.10(-2) s(-1) and a Hill factor of 3.3. The apparent K-m value for HYA was estimated to be 98 mu M, with a k(cat) of 1.2.10(-3) s(-1). (C) 2014, The Society for Biotechnology, Japan. All rights reserved.
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