Runquist D, Fonseca C, Rådström P, Spencer-Martins I, Hahn-Hägerdal B
Department of Applied Microbiology, Lund University, P.O. Box 124, 22100, Lund, Sweden.
Appl Microbiol Biotechnol. 2009 Feb;82(1):123-30. doi: 10.1007/s00253-008-1773-y. Epub 2008 Nov 12.
The glucose/xylose facilitator Gxf1 from Candida intermedia was expressed in the recombinant xylose-fermenting Saccharomyces cerevisiae strain TMB 3057. The new strain, TMB 3411, displayed approximately two times lower K (m) for xylose transport compared to a control strain not expressing Gxf1. In aerobic batch cultivation, the specific growth rate was significantly higher at low xylose concentration, 4 g/L, when Gxf1 was expressed, whereas it remained unchanged at high xylose concentration, 40 g/L. Similarly, in aerobic-xylose-limited chemostat culture, the Gxf1-expressing strain consumed more xylose than the control strain at low dilution rates (low xylose concentration), whereas the situation was reversed at higher dilution rates (high xylose concentration). Also, under anaerobic conditions, the Gxf1-expressing strain showed faster xylose uptake and ethanol formation at low substrate concentrations. The results are discussed in relation to previous observations, which suggested that transport controlled xylose utilization in recombinant xylose-utilizing S. cerevisiae only at low xylose concentrations.
来自中间假丝酵母的葡萄糖/木糖转运蛋白Gxf1在重组木糖发酵酿酒酵母菌株TMB 3057中表达。与未表达Gxf1的对照菌株相比,新菌株TMB 3411对木糖转运的米氏常数(Km)低约两倍。在好氧分批培养中,当表达Gxf1时,在低木糖浓度(4 g/L)下比生长速率显著更高,而在高木糖浓度(40 g/L)下保持不变。同样,在好氧木糖限制的恒化器培养中,在低稀释率(低木糖浓度)下,表达Gxf1的菌株比对照菌株消耗更多的木糖,而在较高稀释率(高木糖浓度)下情况相反。此外,在厌氧条件下,表达Gxf1的菌株在低底物浓度下显示出更快的木糖摄取和乙醇形成。结合先前的观察结果对这些结果进行了讨论,先前的观察结果表明,转运仅在低木糖浓度下控制重组利用木糖的酿酒酵母中的木糖利用。