Мolecular-genetic analysis of determinants encoding β-galactosidases of bacteria Bifidobacterium longum BIM B-813 D

A. N. Morozova, A. Akhremchuk, N. Golovnyova
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Abstract

The molecular-genetic analysis of the bacterial genome of the strain B. longum BIM B-813D distinguished by a high level of β-galactosidase production was performed. Genes Bgal_small_N, lacZ1, bgaB1, bgaB2 and bgaB3, and lacZ2, encoding the synthesis of β-galactosidases, were revealed in the deciphered genome. It was shown that the genes lacZ1, bgaB2, and bgaB3 characterized by an enhanced degree of similarity to the genes of closely related bifidobacterial species, presumably code for the enzymes catalyzing the specific reactions of hydrolysis and transglycosylation of carbohydrates. It was found that the enzymes BgaB1, BgaB2 and BgaB3 belong to the GH42 family of glycosyl hydrolases, whereas the enzymes LacZ1 and LacZ2 – to the GH2 family. The genome domains responsible for the synthesis of β-galactosidases in the strain B. longum BIM B-813D were studied in detail. A comparative analysis of the locus of lacZ1 in B. longum BIM B-813D and the similar genome fragment AS143_01230 from B. longum subsp. longum MC-42 detected the presence of the transposase gene ISL3 in the former strain. It was suggested that the insertion of the sequence of ISL3 in the lacZ1 locus resulted in the modified gene expression and the increased production of β-galactosidase in the strain B. longum BIM B-813D.
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Мolecular-genetic长双歧杆菌BIM b - 813d β-半乳糖苷酶编码决定因素分析
对菌株B. longum BIM B-813D的细菌基因组进行了分子遗传学分析,该菌株具有高水平的β-半乳糖苷酶生产。基因Bgal_small_N、lacZ1、bgaB1、bgaB2、bgaB3和lacZ2编码β-半乳糖苷酶的合成。结果表明,基因lacZ1、bgaB2和bgaB3与近缘双歧杆菌物种的基因相似性增强,可能是催化碳水化合物水解和转糖基化特定反应的酶的编码。结果表明,BgaB1、BgaB2和BgaB3酶属于GH42糖基水解酶家族,而LacZ1和LacZ2 -酶属于GH2家族。详细研究了B. longum BIM B-813D菌株中负责合成β-半乳糖苷酶的基因组结构域。长叶蝉BIM B-813D中lacZ1位点与长叶蝉AS143_01230基因组片段的比较分析。longum MC-42检测到前者菌株中存在转座酶基因ISL3。结果表明,在菌株B. longum BIM B-813D的lacZ1位点插入ISL3序列,导致了该基因表达的改变和β-半乳糖苷酶产量的增加。
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