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Characterization and Functional Analysis of the MAL and MPH Loci for Maltose Utilization in Some Ale and Lager Yeast Strains

机译:某些啤酒和啤酒酵母中麦芽糖利用的MAL和MPH基因座的表征和功能分析

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摘要

Maltose and maltotriose are the major sugars in brewer's wort. Brewer's yeasts contain multiple genes for maltose transporters. It is not known which of these express functional transporters. We correlated maltose transport kinetics with the genotypes of some ale and lager yeasts. Maltose transport by two ale strains was strongly inhibited by other α-glucosides, suggesting the use of broad substrate specificity transporters, such as Agt1p. Maltose transport by three lager strains was weakly inhibited by other α-glucosides, suggesting the use of narrow substrate specificity transporters. Hybridization studies showed that all five strains contained complete MAL1, MAL2, MAL3, and MAL4 loci, except for one ale strain, which lacked a MAL2 locus. All five strains also contained both AGT1 (coding a broad specificity α-glucoside transporter) and MAL11 alleles. MPH genes (maltose permease homologues) were present in the lager but not in the ale strains. During growth on maltose, the lager strains expressed AGT1 at low levels and MALx1 genes at high levels, whereas the ale strains expressed AGT1 at high levels and MALx1 genes at low levels. MPHx expression was negligible in all strains. The AGT1 sequences from the ale strains encoded full-length (616 amino acid) polypeptides, but those from both sequenced lager strains encoded truncated (394 amino acid) polypeptides that are unlikely to be functional transporters. Thus, despite the apparently similar genotypes of these ale and lager strains revealed by hybridization, maltose is predominantly carried by AGT1-encoded transporters in the ale strains and by MALx1-encoded transporters in the lager strains.
机译:麦芽糖和麦芽三糖是啤酒麦芽汁中的主要糖分。布鲁尔酵母含有麦芽糖转运蛋白的多个基因。尚不知道这些表达中的哪些是功能性转运蛋白。我们将麦芽糖运输动力学与一些淡啤酒和啤酒的基因型相关联。两个强麦芽糖菌株的麦芽糖转运受到其他α-葡萄糖苷的强烈抑制,表明使用了广泛的底物特异性转运蛋白,例如Agt1p。三种较大菌株的麦芽糖转运受到其他α-葡萄糖苷的微弱抑制,表明使用了狭窄的底物特异性转运蛋白。杂交研究表明,所有五个菌株均包含完整的MAL1,MAL2,MAL3和MAL4位点,只有一个啤酒菌株缺少MAL2基因座。所有五个菌株还同时含有AGT1(编码广泛特异性α-葡萄糖苷转运蛋白)和MAL11等位基因。 MPH基因(麦芽糖通透酶同系物)存在于啤酒中,而不存在于啤酒中。在麦芽糖上生长期间,较大的菌株低水平表达AGT1,而高水平表达MALx1基因,而强力啤酒菌株高水平表达AGT1,低表达MALx1基因。在所有菌株中,MPHx表达均可忽略不计。啤酒菌株的AGT1序列编码全长(616个氨基酸)多肽,但两个测序啤酒菌株的AGT1序列编码的截短(394个氨基酸)多肽均不太可能是功能性转运蛋白。因此,尽管通过杂交揭示了这些啤酒和啤酒菌株的基因型明显相似,麦芽糖主要由啤酒菌株中的AGT1编码转运蛋白和啤酒菌株中的MALx1编码转运蛋白携带。

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