Efficient production of active form of recombinant cassava hydroxynitrile lyase using <Emphasis Type="Italic">Escherichia coli</Emphasis> in low-temperature culture |
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Authors: | Hisashi Semba Eita Ichige Tadayuki Imanaka Haruyuki Atomi Hideki Aoyagi |
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Institution: | (1) Strategic Technology Research Center, Nippon Shokubai Co., Ltd, 1-25-12, Kannondai, Tsukuba Ibaraki, 305-0856, Japan;(2) Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto 606-8501, Japan;(3) Graduate School of Life and Environmental Sciences, Tsukuba University, Tsukuba Ibaraki, 305-8572, Japan |
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Abstract: | Overexpression and production of the high concentration of hydroxynitrile lyase from cassava (Manihot esculenta (MeHNL, EC 4.1.2.39)) were investigated. Hydroxynitrile lyase is a useful enzyme for the production of optically active cyanohydrin
compounds. The production of MeHNL was increased by changing the rare codons of the original sequence of cassava MeHNL. However,
most of the produced MeHNL was in the insoluble form. In order to increase the solubility of MeHNL, the effects of the cultivation
temperature were investigated. When the cultivation temperature was reduced, the cell yield and the ratio of soluble MeHNL
increased significantly. The enzyme activity and yield at low-temperature cultures (17 °C) were 850 times higher than those
obtained at the optimum growth temperature of 37 °C. The rate of MeHNL production in the present study was calculated as 3,000 unit/h.
Low-temperature cultivation was very effective in improving the productivity of the active form of MeHNL. Unlike the temperature-shift
method, low-temperature cultivation has more potential for the large-scale production of MeHNL for the optically active cyanohydrin
production.
An erratum to this article can be found at |
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Keywords: | Hydroxynitrile lyase Recombinant enzyme High-density culture Low temperature cultivation Cyanohydrin |
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