Lyutova L V, Naumov G I, Shnyreva A V, Naumova E S
State Research Institute of Genetics and Selection of Industrial Microorganisms, National Research Center "Kurchatov Institute," Moscow, 117545 Russia.
Faculty of Biology, Moscow State University, Moscow, 119234 Russia.
Mol Biol (Mosk). 2021 Jan-Feb;55(1):75-85. doi: 10.31857/S0026898421010109.
The ability to ferment lactose is a characteristic peculiarity of dairy Kluyveromyces lactis yeasts; the vast majority of other yeast species are not able to assimilate this disaccharide. Molecular polymorphism of LAC4 genes encoding β-galactosidase controlling lactose fermentation is not well studied, and the published data concern only a single strain (K. lactis var. lactis NRRL Y-1140) isolated from cream in the United States. We studied β-galactosidase genes in lactose-fermenting К lactis strains isolated from dairy products and natural sources in different regions of the world using molecular karyotyping, Southern hybridization, and sequencing. It was established that the ability to ferment lactose in К. lactis var. lactis dairy yeasts is controlled by at least three polymeric LAC loci with different chromosomal localization: LAC1 (chromosome III), LAC2 (II), and LAC3 (IV). Most of the strains we studied had the LAC2 locus. A comparative analysis of β-galactosidases of the Kluyveromyces genus yeasts and these enzymes from other yeasts was conducted for the first time. Phylogenetic analysis detected significant differences between the LAC4 proteins of yeasts of the Kluyveromyces genus (K. lactis, К. marxianus, К. aestuarii, К. nonfermentans, К. wickerhamii), Scheffersomyces stipitis, Sugiyamaella lignohabitans, and Debaryomyces hansenii. A correlation between β-galactosidase sequences and ecological origin (dairy products and natural sources) of Kluyveromyces strains was found. The group of dairy strains is heterogeneous and includes К. lactis var. lactis and К. marxianus yeasts (99.80-100% similarity), which indicates a common origin of their LAC4 genes. Phylogenetic analysis of β-galactosidases indicates a close genetic relationship of dairy and hospital strains of К. lactis var. lactis and К. marxianus. Clinical isolates are able to ferment lactose and appear to originate from the dairy yeasts.
发酵乳糖的能力是乳制品乳酸克鲁维酵母的一个独特特性;绝大多数其他酵母物种无法同化这种二糖。编码控制乳糖发酵的β-半乳糖苷酶的LAC4基因的分子多态性尚未得到充分研究,已发表的数据仅涉及从美国奶油中分离出的单个菌株(乳酸克鲁维酵母乳酸变种NRRL Y-1140)。我们使用分子核型分析、Southern杂交和测序技术,研究了从世界不同地区的乳制品和自然来源中分离出的乳糖发酵乳酸克鲁维酵母菌株中的β-半乳糖苷酶基因。已确定,乳酸克鲁维酵母乳酸变种乳制品酵母中发酵乳糖的能力至少由三个具有不同染色体定位的聚合LAC基因座控制:LAC1(染色体III)、LAC2(II)和LAC3(IV)。我们研究的大多数菌株都有LAC2基因座。首次对克鲁维酵母属酵母的β-半乳糖苷酶与其他酵母的这些酶进行了比较分析。系统发育分析检测到克鲁维酵母属酵母(乳酸克鲁维酵母、马克斯克鲁维酵母、河口克鲁维酵母、非发酵克鲁维酵母、威克汉姆克鲁维酵母)、树干毕赤酵母、木栖 sugiyamaella和汉逊德巴利酵母的LAC4蛋白之间存在显著差异。发现了克鲁维酵母菌株的β-半乳糖苷酶序列与其生态起源(乳制品和自然来源)之间的相关性。乳制品菌株组是异质的,包括乳酸克鲁维酵母乳酸变种和马克斯克鲁维酵母(99.80-100%相似性),这表明它们的LAC4基因有共同的起源。β-半乳糖苷酶的系统发育分析表明,乳酸克鲁维酵母乳酸变种和马克斯克鲁维酵母的乳制品菌株和医院菌株之间存在密切的遗传关系。临床分离株能够发酵乳糖,似乎起源于乳制品酵母。