Vidgren Virve, Huuskonen Anne, Virtanen Hannele, Ruohonen Laura, Londesborough John
VTT Technical Research Centre of Finland, P.O. Box 1000, FI-02044 VTT, Finland.
Appl Environ Microbiol. 2009 Apr;75(8):2333-45. doi: 10.1128/AEM.01558-08. Epub 2009 Jan 30.
The use of more concentrated, so-called high-gravity and very-high-gravity (VHG) brewer's worts for the manufacture of beer has economic and environmental advantages. However, many current strains of brewer's yeasts ferment VHG worts slowly and incompletely, leaving undesirably large amounts of maltose and especially maltotriose in the final beers. alpha-Glucosides are transported into Saccharomyces yeasts by several transporters, including Agt1, which is a good carrier of both maltose and maltotriose. The AGT1 genes of brewer's ale yeast strains encode functional transporters, but the AGT1 genes of the lager strains studied contain a premature stop codon and do not encode functional transporters. In the present work, one or more copies of the AGT1 gene of a lager strain were repaired with DNA sequence from an ale strain and put under the control of a constitutive promoter. Compared to the untransformed strain, the transformants with repaired AGT1 had higher maltose transport activity, especially after growth on glucose (which represses endogenous alpha-glucoside transporter genes) and higher ratios of maltotriose transport activity to maltose transport activity. They fermented VHG (24 degrees Plato) wort faster and more completely, producing beers containing more ethanol and less residual maltose and maltotriose. The growth and sedimentation behaviors of the transformants were similar to those of the untransformed strain, as were the profiles of yeast-derived volatile aroma compounds in the beers.
使用浓度更高的所谓高比重和超高比重(VHG)啤酒麦芽汁来酿造啤酒具有经济和环境优势。然而,目前许多啤酒酵母菌株发酵VHG麦芽汁的速度缓慢且不完全,导致最终啤酒中残留大量不理想的麦芽糖,尤其是麦芽三糖。α-葡萄糖苷通过多种转运蛋白进入酿酒酵母,包括Agt1,它是麦芽糖和麦芽三糖的良好载体。啤酒艾尔酵母菌株的AGT1基因编码功能性转运蛋白,但所研究的拉格酵母菌株的AGT1基因含有一个提前终止密码子,不编码功能性转运蛋白。在本研究中,用艾尔酵母菌株的DNA序列修复了拉格酵母菌株的一个或多个AGT1基因拷贝,并置于组成型启动子的控制下。与未转化菌株相比,修复后的AGT1转化子具有更高的麦芽糖转运活性,尤其是在葡萄糖上生长后(葡萄糖会抑制内源性α-葡萄糖苷转运蛋白基因),并且麦芽三糖转运活性与麦芽糖转运活性的比率更高。它们发酵VHG(24°柏拉图)麦芽汁的速度更快且更完全,产生的啤酒含有更多乙醇,残留的麦芽糖和麦芽三糖更少。转化子的生长和沉降行为与未转化菌株相似,啤酒中酵母衍生的挥发性香气化合物谱也相似。