caa a22 4500 605499160 CHVBK 20210128100549.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6323-1 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6323-1 A third glucose uptake bypass in Corynebacterium glutamicum ATCC 31833 [Elektronische Daten] [Masato Ikeda, Norio Noguchi, Masakazu Ohshita, Akihiro Senoo, Satoshi Mitsuhashi, Seiki Takeno] In Corynebacterium glutamicum, the phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS) has long been the only known glucose uptake system, but we recently found suppressor mutants emerging from a PTS-negative strain of C. glutamicum ATCC 31833 on glucose agar plates, and identified two alternative potential glucose uptake systems, the myo-inositol transporters encoded by iolT1 and iolT2. The expression of either gene renders the PTS-negative strain WTΔptsH capable of growing on glucose. In the present study, we found a suppressor strain that still grew on glucose even after the iolT1 and iolT2 genes were both disrupted under the PTS-negative background. Whole-genome sequencing of the suppressor strain SPH1 identified a G-to-T exchange at 134bp upstream of the bglF gene encoding an EII component of the β-glucoside-PTS, which is found in limited wild-type strains of C. glutamicum. Introduction of the mutation into strain WTΔptsH allowed the PTS-negative strain to grow on glucose. Reverse transcription-quantitative PCR analysis revealed that the mutation upregulates the bglF gene by approximately 11-fold. Overexpression of bglF under the gapA promoter in strain WTΔptsH rendered the strain capable of growing on glucose, and deletion of bglF in strain SPH1 abolished the growth again, proving that bglF is responsible for glucose uptake in the suppressor strain. Simultaneous disruption of three glucokinase genes, glk (Cgl2185, NCgl2105), ppgK (Cgl1910, NCgl1835), and Cgl2647 (NCgl2558), in strain SPH1 resulted in no growth on glucose. Plasmid-mediated expression of any of the three genes in the triple-knockout mutant restored the growth on glucose. These results indicate that C. glutamicum ATCC 31833 has an additional non-PTS glucose uptake route consisting of the bglF-specified EII permease and native glucokinases. Springer-Verlag Berlin Heidelberg, 2014 Corynebacterium glutamicum nationallicence Phosphotransferase system nationallicence Glucose uptake bypass nationallicence β-Glucoside-PTS nationallicence bglF nationallicence Ikeda Masato Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut Noguchi Norio Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut Ohshita Masakazu Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut Senoo Akihiro Bioprocess Development Center, Kyowa Hakko Bio Co., Ltd., 305-0841, Tsukuba, Ibaraki, Japan aut Mitsuhashi Satoshi Bioprocess Development Center, Kyowa Hakko Bio Co., Ltd., 305-0841, Tsukuba, Ibaraki, Japan aut Takeno Seiki Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2741-2750 0175-7598 99:6<2741 2015 99 253 https://doi.org/10.1007/s00253-014-6323-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6323-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ikeda Masato Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut NATIONALLICENCE 700 1- Noguchi Norio Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut NATIONALLICENCE 700 1- Ohshita Masakazu Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut NATIONALLICENCE 700 1- Senoo Akihiro Bioprocess Development Center, Kyowa Hakko Bio Co., Ltd., 305-0841, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Mitsuhashi Satoshi Bioprocess Development Center, Kyowa Hakko Bio Co., Ltd., 305-0841, Tsukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Takeno Seiki Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 399-4598, Nagano, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2741-2750 0175-7598 99:6<2741 2015 99 253