The ncgl1108 (PheP (Cg)) gene encodes a new L-Phe transporter in Corynebacterium glutamicum |
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Authors: | Zhao Zhi Ding Jiu-Yuan Li Tang Zhou Ning-Yi Liu Shuang-Jiang |
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Institution: | (1) State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Chaoyang District, Beijing, 100101, People’s Republic of China;(2) Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Chaoyang District, Beijing, 100101, People’s Republic of China;(3) Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, People’s Republic of China;(4) Institute of Microbiology, Chinese Academy of Sciences, Beichen-Xilu, Chaoyang District, Beijing, 100101, People’s Republic of China; |
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Abstract: | Corynebacterium glutamicum played a central role in the establishment of fermentative production of amino acids, and it is a model for genetic and physiological
studies. The general aromatic amino acid transporter, AroP
Cg
, was the sole functionally identified aromatic amino acid transporter from C. glutamicum. In this study, the ncgl1108 (named as pheP
Cg
), which is located upstream of the genetic cluster (ncgl1110 ∼ ncgl1113) for resorcinol catabolism, was identified as a new l-Phe specific transporter from C. glutamicum RES167. The disruption of pheP
Cg
resulted in RES167∆ncgl1108, and this mutant showed decreased growth on l-Phe (as nitrogen source) but not on l-Tyr or l-Trp. Uptake assays with unlabeled and 14C-labeled l-Phe and l-Tyr indicated that the mutants RES167∆ncgl1108 showed significant reduction in l-Phe uptake than RES167. Expression of pheP
Cg
in RES167∆ncgl1108/pGXKZ1 or RES167∆(ncgl1108-aroP
Cg
)/pGXKZ1 restored their ability to uptake for l-Phe and growth on l-Phe. The uptake of l-Phe was not inhibited by nine amino acids but by l-Tyr. The K
m and V
max values of RES167∆(ncgl1108-aroP
Cg
)/pGXKZ1 for l-Phe were determined to be 10.4 ± 1.5 μM and 1.2 ± 0.1 nmol min−1 (mg DW)−1, respectively, which are different from K
m and V
max values of RES167∆(ncgl1108-aroP
Cg
) for l-Phe 4.0 ± 0.4 μM and 0.6 ± 0.1 nmol min−1 (mg DW)−1]. In conclusion, this PheP
Cg
is a new l-Phe transporter in C. glutamicum. |
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Keywords: | |
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