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ncgl1108(PheP(Cg))基因编码谷氨酸棒杆菌中的一种新型 L-苯丙氨酸转运蛋白。

The ncgl1108 (PheP (Cg)) gene encodes a new L-Phe transporter in Corynebacterium glutamicum.

机构信息

State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Chaoyang District, Beijing, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2011 Jun;90(6):2005-13. doi: 10.1007/s00253-011-3245-z. Epub 2011 Apr 6.

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 (14)C-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.

摘要

谷氨酸棒杆菌在建立氨基酸发酵生产中发挥了核心作用,是遗传和生理研究的模式生物。一般芳香族氨基酸转运蛋白 AroP(Cg) 是谷氨酸棒杆菌中唯一鉴定的功能性芳香族氨基酸转运蛋白。在这项研究中,ncgl1108(命名为 pheP (Cg),位于间苯二酚分解代谢的基因簇(ncgl1110~ncgl1113)上游)被鉴定为谷氨酸棒杆菌 RES167 的新 L-Phe 特异性转运蛋白。pheP (Cg) 的缺失导致 RES167∆ncgl1108 的产生,该突变体在 L-Phe(作为氮源)上的生长受到抑制,但在 L-Tyr 或 L-Trp 上不受抑制。使用未标记和 (14)C 标记的 L-Phe 和 L-Tyr 进行摄取实验表明,突变体 RES167∆ncgl1108 的 L-Phe 摄取量明显低于 RES167。在 RES167∆ncgl1108/pGXKZ1 或 RES167∆(ncgl1108-aroP (Cg))/pGXKZ1 中表达 pheP (Cg) 恢复了它们对 L-Phe 的摄取能力和在 L-Phe 上的生长能力。L-Phe 的摄取不受九种氨基酸的抑制,但受 L-Tyr 的抑制。RES167∆(ncgl1108-aroP (Cg))/pGXKZ1 对 L-Phe 的 K (m) 和 V (max) 值分别为 10.4 ± 1.5 μM 和 1.2 ± 0.1 nmol min(-1) (mg DW)(-1),与 RES167∆(ncgl1108-aroP (Cg)) 对 L-Phe 的 K (m) 和 V (max) 值[4.0 ± 0.4 μM 和 0.6 ± 0.1 nmol min(-1) (mg DW)(-1)]不同。综上所述,该 PheP(Cg) 是谷氨酸棒杆菌中的一种新的 L-Phe 转运蛋白。

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