Diderich J A, Schepper M, van Hoek P, Luttik M A, van Dijken J P, Pronk J T, Klaassen P, Boelens H F, de Mattos M J, van Dam K, Kruckeberg A L
E. C. Slater Institute, University of Amsterdam, Plantage Muidergracht 12, 1018 TV Amsterdam, The Netherlands.
J Biol Chem. 1999 May 28;274(22):15350-9. doi: 10.1074/jbc.274.22.15350.
The kinetics of glucose transport and the transcription of all 20 members of the HXT hexose transporter gene family were studied in relation to the steady state in situ carbon metabolism of Saccharomyces cerevisiae CEN.PK113-7D grown in chemostat cultures. Cells were cultivated at a dilution rate of 0.10 h-1 under various nutrient-limited conditions (anaerobically glucose- or nitrogen-limited or aerobically glucose-, galactose-, fructose-, ethanol-, or nitrogen-limited), or at dilution rates ranging between 0.05 and 0.38 h-1 in aerobic glucose-limited cultures. Transcription of HXT1-HXT7 was correlated with the extracellular glucose concentration in the cultures. Transcription of GAL2, encoding the galactose transporter, was only detected in galactose-limited cultures. SNF3 and RGT2, two members of the HXT family that encode glucose sensors, were transcribed at low levels. HXT8-HXT17 transcripts were detected at very low levels. A consistent relationship was observed between the expression of individual HXT genes and the glucose transport kinetics determined from zero-trans influx of 14C-glucose during 5 s. This relationship was in broad agreement with the transport kinetics of Hxt1-Hxt7 and Gal2 deduced in previous studies on single-HXT strains. At lower dilution rates the glucose transport capacity estimated from zero-trans influx experiments and the residual glucose concentration exceeded the measured in situ glucose consumption rate. At high dilution rates, however, the estimated glucose transport capacity was too low to account for the in situ glucose consumption rate.
研究了酿酒酵母CEN.PK113 - 7D在恒化器培养中生长时,葡萄糖转运动力学以及HXT己糖转运蛋白基因家族所有20个成员的转录情况,这些研究与原位碳代谢的稳态相关。细胞在不同营养限制条件下(厌氧葡萄糖或氮限制,或需氧葡萄糖、半乳糖、果糖、乙醇或氮限制)以0.10 h⁻¹的稀释率进行培养,或者在需氧葡萄糖限制培养中以0.05至0.38 h⁻¹的稀释率进行培养。HXT1 - HXT7的转录与培养物中的细胞外葡萄糖浓度相关。编码半乳糖转运蛋白的GAL2的转录仅在半乳糖限制培养中检测到。HXT家族中编码葡萄糖传感器的两个成员SNF3和RGT2转录水平较低。HXT8 - HXT17的转录本检测水平极低。在单个HXT基因的表达与通过5秒内¹⁴C - 葡萄糖零转运流入量测定的葡萄糖转运动力学之间观察到一致的关系。这种关系与先前在单HXT菌株研究中推导的Hxt1 - Hxt7和Gal2的转运动力学大致相符。在较低稀释率下,从零转运流入实验估计的葡萄糖转运能力和残余葡萄糖浓度超过了测量的原位葡萄糖消耗率。然而,在高稀释率下,估计的葡萄糖转运能力过低,无法解释原位葡萄糖消耗率。