Improving d-mannitol productivity of Escherichia coli: Impact of NAD,CO2 and expression of a putative sugar permease from Leuconostoc pseudomesenteroides |
| |
| Authors: | Florian Heuser Kay Marin Björn Kaup Stephanie Bringer Hermann Sahm |
| |
| Institution: | 1. Forschungszentrum Jülich GmbH, Institute of Biotechnology 1, D-52425 Jülich, Germany;2. University of Cologne, Institute of Biochemistry, Zülpicher Street 47, Germany;1. Department of Chemistry, University of Richmond, Richmond, VA 23173, USA;2. Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC 27599, USA;3. Laboratory for Molecular Sensing, IBP-CNR, Via Pietro Castellino 111, 80131 Napoli, Italy;1. Department of Parasitology and Tropical Medicine, and Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 660-751, Republic of Korea;2. Division of Malaria and Parasitic Diseases, National Institute of Health, Korea Centers for Disease Control and Prevention, Osong 363-951, Republic of Korea;3. Department of Microbiology, Institute of Health Sciences and PMBBRC, Gyeongsang National University School of Medicine, Jinju 660-751, Republic of Korea;4. Department of Tropical Medicine, and Inha Research Institute for Medical Sciences, Inha University School of Medicine, Incheon 400-712, Republic of Korea;1. Laboratory of Industrial Chemistry and Reaction Engineering, Åbo Akademi University, FI-20500 Turku, Finland;2. N. N. Vorozhtsov Institute of Organic Chemistry, Russian Academy of Sciences, Novosibirsk 630090, Russia;3. Novosibirsk State University, Novosibirsk 630090, Russia;1. Department of Solid State Engineering, University of Chemistry and Technology Prague, 166 28 Prague, Czech Republic;2. Department of Organic Technology, University of Chemistry and Technology Prague, 166 28 Prague, Czech Republic;3. Faculty of Science, University of J. E. Purkyně, 400 96 Ústí nad Labem, Czech Republic |
| |
| Abstract: | The highly productive whole-cell biotransformation of d-fructose to d-mannitol with recombinant, resting cells of Escherichia coli BL21(DE3) requires the combined expression of mdh, fdh and glf which encode mannitol and formate dehydrogenases and a sugar facilitator, respectively. However, long-term stability of the system was restricted, possibly due to loss of the cofactor NAD, high concentrations of formate, formation of CO2 affecting the internal pH of the cells, accumulation of high intracellular concentrations of d-mannitol, and export of d-mannitol. Downstream of the mdh gene of Leuconostoc pseudomesenteroides, we identified an open reading frame encoding for a putative mannitol permease. The gene was cloned and expressed in E. coli. Biochemical analyses revealed an activity as secondary carrier for d-fructose. Therefore, the carrier was named FupL and participation in d-mannitol transport was excluded. In biotransformation experiments, the productivity of d-mannitol formation obtained with the strain expressing the additional fupL gene was enhanced by 20%. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
