College of Food and Biological Engineering (Liquor College), Henan University of Animal Husbandry and Economy, Zhengzhou, Henan Province, China.
Postdoctoral Programme, Henan Yangshao Distillery Co., Ltd., Mianchi, Henan Province, China.
PLoS One. 2022 Apr 26;17(4):e0264677. doi: 10.1371/journal.pone.0264677. eCollection 2022.
A total of 296 strains of protease-producing bacteria were isolated and purified from medium-temperature Daqu produced by ZhangGong LaoJiu Wine Co. Ltd. After calculating the ratio of transparent ring diameter to colony diameter and measuring the protease activities, a strain of high-yield protease bacteria, called DW-7, was screened out with a protease activity of 99.54 U/mL. Through morphological observation, 16S rDNA sequence analysis, and physiological and biochemical tests, the isolated bacteria DW-7 was determined to be Bacillus velezensis. In addition, whole-genome sequencing (WGS), using PacBio and the Illumina platform, was performed. Gene annotation was then conducted using the Clusters of Orthologous Groups (COG), Kyoto Encyclopedia of Genes and Genomes (KEGG), Non-Redundant Protein Sequence Database (NR), and Gene Ontology (GO) databases. The results showed that the genome of DW-7 was 3,942,829 bp long with a GC content of 46.45%. A total of 3,662 protein-encoding genes were predicted, with a total length of 3,402,822 bp. Additionally, 2,283; 2,796; and 2,127 genes were annotated in the COG, KEGG, and GO databases, respectively. A total of 196 high-yield protease genes were mainly enriched in the metabolism of alanine, aspartic acid, glutamate, glycine, serine, and threonine, as well as ABC transporter and transporter pathways.
从张弓老酒股份有限公司生产的中温大曲中分离和纯化了 296 株产蛋白酶细菌。通过计算透明环直径与菌落直径的比值，并测量蛋白酶活性，筛选出一株产蛋白酶活性为 99.54 U/mL 的高产蛋白酶细菌 DW-7。通过形态观察、16S rDNA 序列分析和生理生化试验，确定分离得到的 DW-7 菌株为解淀粉芽孢杆菌。此外，还进行了全基因组测序（WGS），使用 PacBio 和 Illumina 平台进行。然后使用 COG、KEGG、NR 和 GO 数据库对基因进行注释。结果表明，DW-7 的基因组大小为 3942829 bp，GC 含量为 46.45%。共预测到 3662 个编码蛋白的基因，总长度为 3402822 bp。此外，在 COG、KEGG 和 GO 数据库中分别注释了 2283、2796 和 2127 个基因。共鉴定到 196 个高产蛋白酶基因，主要富集在丙氨酸、天冬氨酸、谷氨酸、甘氨酸、丝氨酸和苏氨酸代谢以及 ABC 转运体和转运体途径中。
