Environmental Biochemistry Division, Research Center for Inland Seas, Kobe University, Japan.
J Biosci Bioeng. 2009 Sep;108(3):205-10. doi: 10.1016/j.jbiosc.2009.03.020.
Activity of proline uptake in Escherichia coli CSH4 was inhibited in the presence of 1 M NaCl, while it was recovered if the cells were incubated at 30 degrees C for 1 h in a moderate salinity stress (MSS) solution which consists of Davis minimal medium with 5 mM proline and 0.5 M NaCl. Then, an attempt was made to examine whether MSS treatment is also effective on the activity restoration of proline uptake and growth under high salinity for E. coli CSH4 mutants with different combinations of proP, putA, putP, and proU which are related to the transport and metabolization of proline. After MSS treatment, proline uptake was vigorously occurred for the mutants with proline transporter gene proP but not for its deficient ones. For the expression of proline uptake activities of these mutant strains after MSS treatment, PO(4)(3-) in MSS solution is more important than K(+). No growth of strain CSH4 and its mutants without MSS treatment was observed, when cultured in high osmotic medium G (0.8 M NaCl) consisting of 1 mM glycine betaine and Davis minimal medium without potassium phosphate supplemented. After MSS treatment, however, mutant strains lacking proP showed sufficient growth in medium G. Cell growth of proP(+) strains was recognized if MSS treatment was performed in the absence of proline. In conclusion, growth of mutant strains under high-salinity medium G depended on their amount of proline accumulated during MSS treatment, in which K(+) and PO(4)(3-) might play a key role to guarantee their sufficient growth.
在 1 M NaCl 存在的情况下,大肠杆菌 CSH4 的脯氨酸摄取活性受到抑制,而如果细胞在 30°C 下于包含 Davis 基本培养基、5 mM 脯氨酸和 0.5 M NaCl 的中等盐度应激(MSS)溶液中孵育 1 小时,则可恢复其活性。然后,尝试检查 MSS 处理是否对具有不同脯氨酸转运体基因 proP、putA、putP 和 proU 组合的大肠杆菌 CSH4 突变体的脯氨酸摄取活性和高盐条件下的生长恢复也有效,这些基因与脯氨酸的运输和代谢有关。经过 MSS 处理后,具有脯氨酸转运体基因 proP 的突变体能够强烈地摄取脯氨酸,而其缺陷型则不能。对于这些突变菌株在 MSS 处理后的脯氨酸摄取活性表达,MSS 溶液中的 PO(4)(3-)比 K(+)更重要。在不含钾磷酸盐的 1 mM 甘氨酸甜菜碱和 Davis 基本培养基组成的高渗培养基 G(0.8 M NaCl)中,未经 MSS 处理的 CSH4 及其突变株均无法生长。然而,经过 MSS 处理后,缺乏 proP 的突变株在培养基 G 中能够充分生长。如果在没有脯氨酸的情况下进行 MSS 处理,则可以识别 proP(+)菌株的细胞生长。总之,突变株在高盐培养基 G 下的生长取决于它们在 MSS 处理过程中积累的脯氨酸量,其中 K(+)和 PO(4)(3-)可能起到关键作用,以保证它们的充分生长。