Berthels Nele J, Cordero Otero Ricardo R, Bauer Florian F, Pretorius Isak S, Thevelein Johan M
Institute for Wine Biotechnology, Department of Viticulture & Oenology, Stellenbosch University, Victoria Street, Stellenbosch, ZA-7600, South Africa.
Appl Microbiol Biotechnol. 2008 Jan;77(5):1083-91. doi: 10.1007/s00253-007-1231-2. Epub 2007 Oct 23.
Grape juice contains about equal amounts of glucose and fructose, but wine strains of Saccharomyces cerevisiae ferment glucose slightly faster than fructose, leading to fructose concentrations that exceed glucose concentrations in the fermenting must. A high fructose/glucose ratio may contribute to sluggish and stuck fermentations, a major problem in the global wine industry. We evaluated wine yeast strains with different glucose and fructose consumption rates to show that a lower glucose preference correlates with a higher fructose/glucose phosphorylation ratio in cell extracts and a lower K (m) for both sugars. Hxk1 has a threefold higher V (max) with fructose than with glucose, whereas Hxk2 has only a slightly higher V (max) with glucose than with fructose. Overexpression of HXK1 in a laboratory strain of S. cerevisiae (W303-1A) accelerated fructose consumption more than glucose consumption, but overexpression in a wine yeast strain (VIN13) reduced fructose consumption less than glucose consumption. Results with laboratory strains expressing a single kinase showed that total hexokinase activity is inversely correlated with the glucose/fructose (G/F) discrepancy. The latter has been defined as the difference between the rate of glucose and fructose fermentation. We conclude that the G/F discrepancy in wine yeast strains correlates with the kinetic properties of hexokinase-mediated sugar phosphorylation. A higher fructose/glucose phosphorylation ratio and a lower K (m) might serve as markers in selection and breeding of wine yeast strains with a lower tendency for sluggish fructose fermentation.
葡萄汁中葡萄糖和果糖的含量大致相等,但酿酒酵母的葡萄酒菌株发酵葡萄糖的速度略快于果糖,导致在发酵葡萄汁中果糖浓度超过葡萄糖浓度。高果糖/葡萄糖比率可能导致发酵迟缓及发酵停滞,这是全球葡萄酒行业的一个主要问题。我们评估了具有不同葡萄糖和果糖消耗速率的葡萄酒酵母菌株,结果表明较低的葡萄糖偏好与细胞提取物中较高的果糖/葡萄糖磷酸化比率以及两种糖较低的米氏常数(Km)相关。己糖激酶1(Hxk1)对果糖的最大反应速度(Vmax)比对葡萄糖的高3倍,而己糖激酶2(Hxk2)对葡萄糖的最大反应速度仅比对果糖的略高。在酿酒酵母实验室菌株(W303 - 1A)中过表达HXK1,加速果糖消耗的程度超过葡萄糖消耗,但在葡萄酒酵母菌株(VIN13)中过表达时,果糖消耗减少的程度小于葡萄糖消耗。对表达单一激酶的实验室菌株的研究结果表明,总己糖激酶活性与葡萄糖/果糖(G/F)差异呈负相关。后者被定义为葡萄糖和果糖发酵速率之间的差异。我们得出结论,葡萄酒酵母菌株中的G/F差异与己糖激酶介导的糖磷酸化的动力学特性相关。较高的果糖/葡萄糖磷酸化比率和较低的Km可能作为筛选和培育果糖发酵迟缓倾向较低的葡萄酒酵母菌株的标记。