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一株褐藻胶降解菌 Cobetia sp. cqz5-12 的分离鉴定及其全基因组序列分析。

Isolation, identification, and whole genome sequence analysis of the alginate-degrading bacterium Cobetia sp. cqz5-12.

机构信息

College of Life and Environmental Science, Wenzhou University, Wenzhou, 325035, People's Republic of China.

出版信息

Sci Rep. 2020 Jul 2;10(1):10920. doi: 10.1038/s41598-020-67921-7.

Abstract

Alginate-degrading bacteria or alginate lyases can be used to oligomerize alginate. In this study, an alginate-degrading bacterium with high alginolytic activity was successfully screened by using Sargassum fusiforme sludge. When the strain was grown on a plate containing sodium alginate, the transparent ring diameter (D) was 2.2 cm and the ratio (D/d) of transparent ring diameter to colony diameter (d) was 8.8. After 36 h in culture at a temperature of 28 °C shaken at 150 r/min, the enzymatic activity of the fermentation supernatant reached 160 U/mL, and the enzymatic activity of the bacterial precipitate harvested was 2,645 U/mL. The strain was named Cobetia sp. cqz5-12. Its genome is circular in shape, 4,209,007 bp in size, with a 62.36% GC content. It contains 3,498 predicted coding genes, 72 tRNA genes, and 21 rRNA genes. The functional annotations for the coding genes demonstrated that there were 181 coding genes in the genome related to carbohydrate transport and metabolism and 699 coding genes with unknown functions. Three putative coding genes, alg2107, alg2108 and alg2112, related to alginate degradation were identified by analyzing the carbohydrate active enzyme (CAZy) database. Moreover, proteins Alg2107 and Alg2112 were successfully expressed and exhibited alginate lyase activity.

摘要

褐藻胶降解细菌或褐藻胶裂解酶可用于寡聚化褐藻胶。本研究利用羊栖菜污泥成功筛选到一株具有高褐藻胶裂解活性的褐藻胶降解细菌。当该菌株在含有褐藻酸钠的平板上生长时,透明环直径(D)为 2.2 cm,透明环直径与菌落直径(d)的比值(D/d)为 8.8。在 28°C、150 r/min 摇床培养 36 h 后,发酵上清液的酶活达到 160 U/mL,收获的细菌沉淀的酶活达到 2645 U/mL。该菌株被命名为 Cobetia sp. cqz5-12。其基因组为环状,大小为 4209007 bp,GC 含量为 62.36%。它包含 3498 个预测编码基因、72 个 tRNA 基因和 21 个 rRNA 基因。对编码基因的功能注释表明,基因组中有 181 个编码基因与碳水化合物的运输和代谢有关,有 699 个编码基因功能未知。通过分析碳水化合物活性酶(CAZy)数据库,鉴定出三个与褐藻胶降解相关的假定编码基因 alg2107、alg2108 和 alg2112。此外,成功表达了蛋白 Alg2107 和 Alg2112,并显示出褐藻胶裂解酶活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87eb/7331586/32676a338b41/41598_2020_67921_Fig1_HTML.jpg

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