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Discovery of an l‑Fucono-1,5-lactonase from cog3618 of the Amidohydrolase Superfamily

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journal contribution
posted on 08.01.2013, 00:00 by Merlin Eric Hobbs, Matthew Vetting, Howard J. Williams, Tamari Narindoshvili, Devon M. Kebodeaux, Brandan Hillerich, Ronald D. Seidel, Steven C. Almo, Frank M. Raushel
A member of the amidohydrolase superfamily, BmulJ_04915 from Burkholderia multivorans, of unknown function was determined to hydrolyze a series of sugar lactones: l-fucono-1,4-lactone, d-arabino-1,4-lactone, l-xylono-1,4-lactone, d-lyxono-1,4-lactone, and l-galactono-1,4-lactone. The highest activity was shown for l-fucono-1,4-lactone with a kcat value of 140 s–1 and a kcat/Km value of 1.0 × 105 M–1 s–1 at pH 8.3. The enzymatic product of an adjacent l-fucose dehydrogenase, BmulJ_04919, was shown to be l-fucono-1,5-lactone via nuclear magnetic resonance spectroscopy. l-Fucono-1,5-lactone is unstable and rapidly converts nonenzymatically to l-fucono-1,4-lactone. Because of the chemical instability of l-fucono-1,5-lactone, 4-deoxy-l-fucono-1,5-lactone was enzymatically synthesized from 4-deoxy-l-fucose using l-fucose dehydrogenase. BmulJ_04915 hydrolyzed 4-deoxy-l-fucono-1,5-lactone with a kcat value of 990 s–1 and a kcat/Km value of 8.0 × 106 M–1 s–1 at pH 7.1. The protein does not require divalent cations in the active site for catalytic activity. BmulJ_04915 is the second enzyme from cog3618 of the amidohydrolase superfamily that does not require a divalent metal for catalytic activity. BmulJ_04915 is the first enzyme that has been shown to catalyze the hydrolysis of either l-fucono-1,4-lactone or l-fucono-1,5-lactone. The structures of the fuconolactonase and the fucose dehydrogenase were determined by X-ray diffraction methods.