Identification and characterization of γ-aminobutyric acid uptake system GabPCg (NCgl0464) in Corynebacterium glutamicum.
Corynebacterium glutamicum is widely used for industrial production of various amino acids and vitamins, and there is growing interest in engineering this bacterium for more commercial bioproducts such as γ-aminobutyric acid (GABA). In this study, a C. glutamicum GABA-specific transporter (GabP(Cg)) encoded by ncgl0464 was identified and characterized. GabP(Cg) plays a major role in GABA uptake and is essential to C. glutamicum growing on GABA. GABA uptake by GabP(Cg) was weakly competed by l-Asn and l-Gln and stimulated by sodium ion (Na(+)). The K(m) and V(max) values were determined to be 41.1 ± 4.5 μM and 36.8 ± 2.6 nmol min(-1) (mg dry weight [DW])(-1), respectively, at pH 6.5 and 34.2 ± 1.1 μM and 67.3 ± 1.0 nmol min(-1) (mg DW)(-1), respectively, at pH 7.5. GabP(Cg) has 29% amino acid sequence identity to a previously and functionally identified aromatic amino acid transporter (TyrP) of Escherichia coli but low identities to the currently known GABA transporters (17% and 15% to E. coli GabP and Bacillus subtilis GabP, respectively). The mutant RES167 Δncgl0464/pGXKZ9 with the GabP(Cg) deletion showed 12.5% higher productivity of GABA than RES167/pGXKZ9. It is concluded that GabP(Cg) represents a new type of GABA transporter and is potentially important for engineering GABA-producing C. glutamicum strains.
State Key Laboratory of Microbial Resourcesa and Department of Industrial Microbiology and Biotechnology, Beijing, People’s Republic of China.
Zhi
Z
Zhao
Jiu-Yuan
JY
Ding
Wen-Hua
WH
Ma
Ning-Yi
NY
Zhou
Shuang-Jiang
SJ
Liu
D001412:Bacillus subtilis / Q000235:genetics; D001692:Biological Transport; D048230:Corynebacterium glutamicum / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D017353:Gene Deletion; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D026901:Membrane Transport Proteins; D005680:gamma-Aminobutyric Acid