Pokorný Richard, Hudecová Daniela, Burgstaller Wolfgang, Varecka L'udovít
Department of Biochemistry and Microbiology, Slovak University of Technology, Bratislava, Slovak Republic.
FEMS Microbiol Lett. 2004 Jan 15;230(1):123-8. doi: 10.1016/S0378-1097(03)00884-X.
The U-(14)C-labelled glutamate uptake was measured in both sucrose- and glutamate-grown mycelia of Trichoderma viride. The biomass yield was five-fold lower with glutamate as a sole carbon source. The rate of glutamate transport measured at a glutamate concentration of 1 mM remained unchanged in glutamate-grown mycelia whereas the properties of the glutamate transport were substantially changed compared to sucrose-grown mycelia. The glutamate uptake in both sucrose- and glutamate-grown mycelia was inhibited by an uncoupler (3,3',4',5-tetrachlorosalicylanilide) but the inhibitory efficiency was higher in the latter. The affinity of the permease to glutamate increased approximately five-fold in the glutamate-grown mycelia (about 76 microM compared to about 16 microM). The pH optimum for glutamate uptake was 4 in sucrose-grown mycelia but the glutamate-grown mycelia had two pH optima, one at pH 4 and the second between pH 6 and 7. The inhibition of glutamate uptake by other amino acids yielded different inhibitory patterns in the two mycelia under study. The glutamate uptake in mycelia of different ages also showed differences in both transport rate and temporal pattern. The results show that the growth of mycelia on glutamate led to the appearance of an additional permease with different properties and suggest that only this permease is operating in mycelia grown on glutamate.
在绿色木霉以蔗糖和谷氨酸为碳源生长的菌丝体中,均测定了U-(14)C标记的谷氨酸摄取量。以谷氨酸作为唯一碳源时,生物量产量降低了五倍。在谷氨酸浓度为1 mM时测得的谷氨酸转运速率在以谷氨酸生长的菌丝体中保持不变,而与以蔗糖生长的菌丝体相比,谷氨酸转运的特性发生了显著变化。解偶联剂(3,3',4',5-四氯水杨酰苯胺)抑制了以蔗糖和谷氨酸生长的菌丝体中的谷氨酸摄取,但对后者的抑制效率更高。在以谷氨酸生长的菌丝体中,通透酶对谷氨酸的亲和力增加了约五倍(约76 microM,相比之下约为16 microM)。以蔗糖生长的菌丝体中,谷氨酸摄取的最适pH为4,但以谷氨酸生长的菌丝体有两个最适pH,一个在pH 4,另一个在pH 6至7之间。在研究的两种菌丝体中,其他氨基酸对谷氨酸摄取的抑制产生了不同的抑制模式。不同年龄菌丝体中的谷氨酸摄取在转运速率和时间模式上也存在差异。结果表明,菌丝体在谷氨酸上生长导致出现了一种具有不同特性的额外通透酶,并表明只有这种通透酶在以谷氨酸生长的菌丝体中起作用。