De novo engineering and metabolic flux analysis of inosine biosynthesis in <Emphasis Type="Italic">Bacillus subtilis</Emphasis> |
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Authors: | Haojian Li Guoqiang Zhang Aihua Deng Ning Chen Tingyi Wen |
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Institution: | (1) College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China;(2) Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China;(3) Graduate University of Chinese Academy of Sciences, Beijing, 100049, China; |
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Abstract: | Wild-type B. subtilis strain W168 was de novo engineered for inosine biosynthesis. Inactivation of deoD and purA led to 0.15 ± 0.04 and 6.44 ± 0.39 g inosine/l yields, respectively. The deoD
purA double mutant accumulated 7.6 ± 0.34 g inosine/l, with a 4.7% (w/w) conversion ratio from glucose to inosine. Comparative
metabolic flux analysis revealed that the fluxes from inosine to hypoxanthine and from inosine monophosphate to adenosine
monophosphate in the double mutant decreased to 14.0 and 0.61% of those in the wild-type strain. The major role of purA was demonstrated when inactivation of deoD and purA were found to contribute additively to inosine accumulation. This work is expected to contribute to the improvement of the
fermentative production of purine nucleosides in the microbial industry. |
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