Levan-type FOS production using a Bacillus licheniformis endolevanase |
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| Institution: | 1. Dpto. de Ingenieria celular y Biocatalisis, Instituto de Biotecnología UNAM, Av. Universidad #2001, Col. Chamilpa, C.P. 62210 Cuernavaca, Morelos, Mexico;2. Centro de Investigación, DACS-Universidad Juárez Autónoma de Tabasco, Av. Gregorio Méndez no. 2838-A. Col. Tamulte, CP 86150 Villahermosa, Centro, Tabasco, Mexico;3. Departamento de Sistemas Biológicos, Universidad Autónoma Metropolitana, Unidad Xochimilco, Calzada del Hueso 1100 Villa Quietud, 04960 Coyoacán, Distrito Federal, Mexico;1. Biochemistry Laboratory, Faculty of Sciences of Sfax, PB 802, 3018 Sfax, Tunisia;2. Biotechnology Center of Sfax BP “1177” 3018-Sfax, Tunisia, University of Sfax, Tunisia;1. Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, CP 62240 Cuernavaca, Morelos, Mexico;2. Centro de Investigación, DACS-Universidad Juárez Autónoma de Tabasco, Av. Gregorio Méndez no. 2838-A. Col. Tamulte, CP 86150 Villahermosa, Centro, Tabasco, Mexico;1. Department of Plant and Environmental Sciences, Copenhagen Plant Science Centre, University of Copenhagen, Denmark;2. Department of Food Science, University of Copenhagen, Denmark |
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| Abstract: | In the present work we describe an enzymatic production method to obtain β2-6 fructose oligosaccharides (levan-type FOS) through a sequential reaction in which a bacterial endolevanase is applied to levan produced from sucrose by bacterial levansucrases. A putative gene encoding an endolevanase, designated as LevBl, was identified through a bioinformatics search, isolated from a strain of Bacillus licheniformis IBt1 from our own collection and expressed in Escherichia coli. LevB1 showed a specific activity of 1.8 U/mg protein at 35 °C in 50 mM phosphate buffer pH 6.0. A first order kinetic behavior was found when up to 150 g/L of low molecular weight levan (8.3 kDa) was used as the substrate. The product profile was determined by HPAEC-PAD and consisted of levan-type FOS with a polymerization degree between 2 and 8, with levanbiose as the major product after long reaction times. Yields of 97% of levan-type FOS were obtained when 1.0 U/mL of LevB1 reacted with 100 g/L of levan produced by the levansucrase from Bacillus subtilis. Finally, it was observed that levan-type FOS are efficiently fermented by probiotic lactic acid bacteria. |
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| Keywords: | Levan Levan-type FOS Fructooligosaccharides Endolevanase Levansucrase Prebiotics |
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