Fatty acid production from butter using novel cutinase-displaying yeast |
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| Authors: | Ken Horii Takashi Adachi Takanori Tanino Tsutomu Tanaka Atsushi Kotaka Hiroshi Sahara Tsuneharu Hashimoto Nobuyuki Kuratani Seiji Shibasaki Chiaki Ogino Hideo Noda Yoji Hata Mitsuyoshi Ueda Akihiko Kondo |
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| Institution: | 1. San-eigen F.F.I., Inc, 1-1-11 Sanwa-cho, Toyonaka, Osaka 561-8588, Japan;2. Department of Chemical Science and Engineering, Graduate School of Engineering, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan;3. Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan;4. Research Institute, Gekkeikan Sake Co., Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto 612-8385, Japan;5. Kansai Chemical Engineering Co., Ltd., 1-9-7 Minaminanamatsu, Amagasaki 660-0053, Japan;6. Department of Pharmacy, School of Pharmacy, Hyogo University of Health Sciences, Minatojima, Chuo-ku, Kobe 650-8530, Japan;7. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwake, Sakyo-ku, Kyoto 606-8502, Japan |
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| Abstract: | We used cutinase from the filamentous fungi Aspergillus oryzae to produce dairy flavors. Secretory and displayed forms of cutinase were investigated using salt-free butter, which is composed mostly of triacylglycerides, as the substrate. The secretory form of cutinase, which was produced in recombinant A. oryzae, was suitable for producing butyric acids (16.8 mol%). Also, cutinase displayed on the cell surface of the yeast Saccharomyces cerevisiae as a fusion protein with α-agglutinin released butyric acid at a 2.7-fold rate (45.4 mol%) higher than that of the secreted form. Yeasts carrying two copies of cutinase genes into their chromosomes, which were constructed using the HELOH method, released free fatty acids rapidly and showed 2-fold higher lipase activity compared with yeasts carrying one copy of the cutinase gene. |
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