Muhammad Neak, Avila Forbes, Nedashkovskaya Olga I, Kim Song-Gun
Biological Resource Center/Korean Collection for Type Cultures (KCTC), Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea.
Department of Environmental Biotechnology, KRIBB School of Biotechnology, University of Science and Technology (UST), Daejeon, Republic of Korea.
Front Microbiol. 2023 Dec 14;14:1265676. doi: 10.3389/fmicb.2023.1265676. eCollection 2023.
Three novel strains designated ABR2-5, BKB1-1, and WSW4-B4 belonging to the genus of the phylum were isolated from algae and mud samples collected in the West Sea, Korea. All three strains were enriched for genes encoding up to 216 carbohydrate-active enzymes (CAZymes), which participate in the degradation of agar, alginate, carrageenan, laminarin, and starch. The 16S rRNA sequence similarities among the three novel isolates were 94.0%-94.7%, and against all three existing species in the genus they were 93.6%-97.2%. The genome sizes of the strains ABR2-5, BKB1-1, and WSW4-B4 were 5.5, 4.4, and 5.0 Mb, respectively, and the GC content ranged from 41.1%-42.0%. The average nucleotide identity and the digital DNA-DNA hybridization values of each novel strain within the isolates and all existing species in the genus were in a range of 69.2%-75.5% and 17.7-18.9%, respectively, supporting the creation of three new species. The three novel strains exhibited a distinctive fatty acid profile characterized by elevated levels of iso-C (37.7%-47.4%) and C ω5c (14.4%-22.9%). Specifically, strain ABR2-5 displayed an additional higher proportion of C (13.0%). The polar lipids were phosphatidylethanolamine, unidentified lipids, aminolipids, and glycolipids. Menaquinone-7 was identified as the respiratory quinone of the isolates. A comparative genome analysis was performed using the KEGG, RAST, antiSMASH, CRISPRCasFinder, dbCAN, and dbCAN-PUL servers and CRISPRcasIdentifier software. The results revealed that the isolates harbored many key genes involved in central metabolism for the synthesis of essential amino acids and vitamins, hydrolytic enzymes, carotenoid pigments, and antimicrobial compounds. The KEGG analysis showed that the three isolates possessed a complete pathway of dissimilatory nitrate reduction to ammonium (DNRA), which is involved in the conservation of bioavailable nitrogen within the ecosystem. Moreover, all the strains possessed genes that participated in the metabolism of heavy metals, including arsenic, copper, cobalt, ferrous, and manganese. All three isolated strains contain the class 2 type II subtype C1 CRISPR-Cas system in their genomes. The distinguished phenotypic, chemotaxonomic, and genomic characteristics led us to propose that the three strains represent three novel species in the genus : e sp. nov. (ABR2-5 = KCTC 82990 = JCM 35839), sp. nov. (BKB1-1 = KCTC 82964 = JCM 35840), and sp. nov. (WSW4-B4 = KCTC 82706 = JCM 35841).
从韩国西海采集的藻类和泥浆样本中分离出三种新型菌株,分别命名为ABR2-5、BKB1-1和WSW4-B4,它们属于该门的属。所有这三种菌株都富含编码多达216种碳水化合物活性酶(CAZymes)的基因,这些酶参与琼脂、藻酸盐、卡拉胶、海带多糖和淀粉的降解。这三种新型分离株之间的16S rRNA序列相似性为94.0%-94.7%,与该属中所有三种现有物种的序列相似性为93.6%-97.2%。菌株ABR2-5、BKB1-1和WSW4-B4的基因组大小分别为5.5、4.4和5.0 Mb,GC含量在41.1%-42.0%之间。每个新型菌株与分离株中的其他菌株以及该属中所有现有物种的平均核苷酸同一性和数字DNA-DNA杂交值分别在69.2%-75.5%和17.7-18.9%的范围内,这支持了三个新物种的形成。这三种新型菌株表现出独特的脂肪酸谱,其特征是异-C(37.7%-47.4%)和C ω5c(14.4%-22.9%)水平升高。具体而言,菌株ABR2-5显示出更高比例的C(13.0%)。极性脂质为磷脂酰乙醇胺、未鉴定的脂质、氨基脂质和糖脂。甲萘醌-7被鉴定为分离株的呼吸醌。使用KEGG、RAST、antiSMASH、CRISPRCasFinder、dbCAN和dbCAN-PUL服务器以及CRISPRcasIdentifier软件进行了比较基因组分析。结果表明,这些分离株含有许多参与必需氨基酸和维生素合成、水解酶、类胡萝卜素色素和抗菌化合物的中心代谢的关键基因。KEGG分析表明,这三种分离株具有异化硝酸盐还原为铵(DNRA)的完整途径,该途径参与生态系统中生物可利用氮的保存。此外,所有菌株都拥有参与重金属代谢的基因,包括砷、铜、钴、亚铁和锰。所有三种分离菌株的基因组中都含有2类II型C1亚型CRISPR-Cas系统。这些独特的表型、化学分类学和基因组特征使我们提出,这三种菌株代表该属中的三个新物种:新种(ABR2-5 = KCTC 82990 = JCM 35839)、新种(BKB1-1 = KCTC 82964 = JCM 35840)和新种(WSW4-B4 = KCTC 82706 = JCM 35841)。