Eichler W
Institut für Biochemie I der Universität Heidelberg.
Biol Chem Hoppe Seyler. 1990 Mar;371(3):273-82. doi: 10.1515/bchm3.1990.371.1.273.
Growth of Tetrahymena thermophila in a synthetic nutrient medium with or without the essential amino acid L-arginine was studied in the presence or absence of the arginine metabolites L-citrulline and L-ornithine and the polyamines putrescine, spermidine, and spermine. The effects of the growth conditions on the stimulations of the enzymes of the arginine metabolic and polyamine biosynthetic pathway, arginine deiminase (ADI), citrulline hydrolase (CH), ornithine decarboxylase (ODC), and ornithine-oxo-acid aminotransferase were determined. Tetrahymena cells were unable to grow in the absence of L-arginine and the amino-acid utilization was greatly impaired. None of the metabolites or polyamines was able to substitute for arginine. In the presence of arginine, Tetrahymena cultures grew well and citrulline and ornithine did not alter the growth behaviour in any way. In the presence of putrescine, the lag period was decreased from 3 h to 2 h. Spermidine and spermine acted similar to putrescine but less pronounced. The stimulation of the activity of ADI, the key enzyme of arginine degradation, was absolutely dependent upon the presence of arginine in the medium: in the absence of arginine, the low ADI activity which was present in the cells before inoculation was decreased to zero levels within 30 min. In the presence of arginine, the stimulation of ADI was not altered by citrulline and ornithine but putrescine, spermidine, and spermine decreased ADI-stimulation to half of the control values. The stimulation of CH activity in the presence of arginine was not altered by any added metabolite or polyamine. In the media without arginine, stimulation of CH was greatly reduced, in the presence of ornithine more than in its absence, and even more in the presence of putrescine and spermidine. Stimulation of ODC activity in the presence of arginine was not affected by citrulline and ornithine but in the presence of polyamines it was rapidly decreased to unstimulated levels after an initial ca. 10-fold increase. The "hyperstimulation" of ODC in the absence of free arginine was reduced to normal in the presence of citrulline, the stimulation was decreased even below normal levels in the presence of ornithine and polyamines decreased ODC activity to zero levels. O delta T activity was stimulated more in the presence of arginine than in its absence. In both cases the stimulation was enhanced in the presence of polyamines and only in the absence of arginine--by ornithine.(ABSTRACT TRUNCATED AT 400 WORDS)
在有或没有必需氨基酸L-精氨酸的合成营养培养基中,研究嗜热四膜虫在有或没有精氨酸代谢物L-瓜氨酸和L-鸟氨酸以及多胺腐胺、亚精胺和精胺存在的情况下的生长情况。测定了生长条件对精氨酸代谢和多胺生物合成途径中酶(精氨酸脱亚氨酶(ADI)、瓜氨酸水解酶(CH)、鸟氨酸脱羧酶(ODC)和鸟氨酸-氧代-酸转氨酶)刺激作用的影响。在没有L-精氨酸的情况下,嗜热四膜虫细胞无法生长,氨基酸利用受到极大损害。没有一种代谢物或多胺能够替代精氨酸。在有精氨酸的情况下,嗜热四膜虫培养物生长良好,瓜氨酸和鸟氨酸对生长行为没有任何影响。在有腐胺的情况下,延迟期从3小时缩短到2小时。亚精胺和精胺的作用与腐胺相似,但不太明显。精氨酸降解关键酶ADI活性的刺激绝对依赖于培养基中精氨酸的存在:在没有精氨酸的情况下,接种前细胞中存在的低ADI活性在30分钟内降至零水平。在有精氨酸的情况下,瓜氨酸和鸟氨酸不会改变ADI的刺激作用,但腐胺、亚精胺和精胺会将ADI刺激作用降低到对照值的一半。在有精氨酸的情况下,添加任何代谢物或多胺都不会改变CH活性的刺激作用。在没有精氨酸的培养基中,CH的刺激作用大大降低,在有鸟氨酸的情况下比没有鸟氨酸时更甚,在有腐胺和亚精胺的情况下更是如此。在有精氨酸的情况下,瓜氨酸和鸟氨酸不会影响ODC活性的刺激作用,但在有多胺的情况下,在最初约增加10倍后,ODC活性会迅速降至未刺激水平。在没有游离精氨酸的情况下ODC的“过度刺激”在有瓜氨酸的情况下恢复正常,在有鸟氨酸的情况下刺激作用甚至低于正常水平,多胺会将ODC活性降低到零水平。在有精氨酸的情况下,OδT活性的刺激作用比没有精氨酸时更强。在这两种情况下,多胺的存在都会增强刺激作用,并且仅在没有精氨酸的情况下——鸟氨酸也会增强刺激作用。(摘要截断于400字)