Yamamoto S, Yamasaki K, Takashina K, Katsu T, Shinoda S
Faculty of Pharmaceutical Sciences, Okayama University.
Microbiol Immunol. 1989;33(1):11-21. doi: 10.1111/j.1348-0421.1989.tb01493.x.
Nongrowing Vibrio parahaemolyticus cells rapidly produced putrescine (Put) from added arginine when subjected to a low osmotic stress. This phenomenon was characterized in connection with a regulatory mechanism of the responsible enzymes, arginine decarboxylase (ADC) and agmatine ureohydrolase (AUH). NaCl, KCl, LiCl, sucrose, and glycerol were used as solutes to prepare the resuspending media with various osmolalities. Regardless of whether the solutes were electrolytes or non-electrolytes, exposure of cells to low osmolality brought about instantaneous increases in both intra- and extracellular Put contents without significant changes in the contents of other polyamines. This acceleration in Put production was accompanied by no increases in the specific activities of ADC and AUH. On the other hand, when cells were exposed to the osmolality equivalent to 2 or 5% NaCl, all solutes except for glycerol did not cause a remarkable variation in the intracellular Put content, while the amount of Put in the medium varied depending on the solute used; sucrose and glycerol still greatly prompted Put production, as judged by high Put contents in the media, even at the osmolality equivalent to 5% NaCl. The cation efflux from cells, measured as the K+ release, was observed whenever the increase in Put production occurred. Furthermore, in vitro experiments showed that NaCl and KCl inhibited ADC to a similar extent, about 70% inhibition being observed at 200 mM. However, AUH was not affected by these compounds. These results suggest that the reduction in the concentrations of Na+ and K+ predominantly present in cells may cause the increase in activity of the preexisting ADC, which leads to the enhancement of Put production.
非生长状态的副溶血性弧菌细胞在受到低渗胁迫时,能迅速利用添加的精氨酸产生腐胺(Put)。该现象与负责的酶,即精氨酸脱羧酶(ADC)和胍丁胺脲水解酶(AUH)的调节机制有关。使用氯化钠、氯化钾、氯化锂、蔗糖和甘油作为溶质来制备具有不同渗透压的重悬培养基。无论溶质是电解质还是非电解质,将细胞暴露于低渗环境都会导致细胞内和细胞外Put含量瞬间增加,而其他多胺的含量没有显著变化。Put产量的这种增加伴随着ADC和AUH比活性的没有增加。另一方面,当细胞暴露于相当于2%或5%氯化钠的渗透压时,除甘油外的所有溶质都不会导致细胞内Put含量有显著变化,而培养基中Put的量则根据所用溶质而变化;即使在相当于5%氯化钠的渗透压下,蔗糖和甘油仍能极大地促进Put的产生,从培养基中高Put含量可以判断。每当Put产量增加时,都会观察到细胞阳离子外流,以钾离子释放来衡量。此外,体外实验表明,氯化钠和氯化钾对ADC的抑制程度相似,在200 mM时约有70%的抑制率。然而,AUH不受这些化合物的影响。这些结果表明,细胞中主要存在的钠离子和钾离子浓度的降低可能导致预先存在的ADC活性增加,从而导致Put产量的提高。