Kokoulin Maxim S, Savicheva Yulia V, Otstavnykh Nadezhda Y, Kurilenko Valeria V, Meleshko Dmitry A, Isaeva Marina P
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, 159/2, Prospect 100 let Vladivostoku, Vladivostok 690022, Russia.
Principal Engineering School, ITMO University, 9, Lomonosova Street, St. Petersburg 191002, Russia.
Int J Mol Sci. 2024 Dec 1;25(23):12927. doi: 10.3390/ijms252312927.
sp. KMM 8419 (=CB1-14) is a Gram-negative bacterium isolated from a food-net mucus sample of marine polychaete collected in the Sea of Japan. Here, we report the structure and biosynthetic gene cluster of the capsular polysaccharide (CPS) from strain KMM 8419. The CPS was isolated and studied by one- and two-dimensional H and C nuclear magnetic resonance (NMR) spectroscopy. The molecular weight of the CPS was about 254 kDa. The CPS consisted of disaccharide repeating units of D-glucose and sulfated and acetylated L-rhamnose established as →2)-α-L-Rha3S4Ac-(1→6)-α-D-Glc-(1→. To identify the genes responsible for CPS biosynthesis, whole-genome sequencing of KMM 8419 was carried out. Based on the genome annotations together with the Interproscan, UniProt and AntiSMASH results, a CPS-related gene cluster of 80 genes was found on chromosome 1. This cluster contained sets of genes encoding for the nucleotide sugar biosynthesis (UDP-Glc and dTDP-Rha), assembly (glycosyltransferases (GT)), transport (ABC transporter) and sulfation (PAPS biosynthesis and sulfotransferases) of the sulfated CPS. A hypothetical model for the assembly and transportation of the sulfated CPS was also proposed. In addition, this locus included genes for O-antigen biosynthesis. Further studies of biological activity, the structure-activity relationship in the new sulfated polysaccharide and its biosynthesis are necessary for the development of potent anticancer agents or drug delivery systems.
菌株KMM 8419（=CB1-14）是一种革兰氏阴性菌，从日本海采集的海洋多毛纲动物的食物网黏液样本中分离得到。在此，我们报道了菌株KMM 8419的荚膜多糖（CPS）的结构和生物合成基因簇。通过一维和二维氢及碳核磁共振（NMR）光谱对CPS进行了分离和研究。CPS的分子量约为254 kDa。CPS由D-葡萄糖以及硫酸化和乙酰化的L-鼠李糖的二糖重复单元组成，确定为→2)-α-L-Rha3S4Ac-(1→6)-α-D-Glc-(1→。为了鉴定负责CPS生物合成的基因，对KMM 8419进行了全基因组测序。基于基因组注释以及Interproscan、UniProt和AntiSMASH的结果，在1号染色体上发现了一个由80个基因组成的与CPS相关的基因簇。该簇包含编码硫酸化CPS的核苷酸糖生物合成（UDP-葡萄糖和dTDP-鼠李糖）、组装（糖基转移酶（GT））、转运（ABC转运蛋白）和硫酸化（PAPS生物合成和磺基转移酶）的基因集。还提出了硫酸化CPS组装和转运的假设模型。此外，该位点还包括O抗原生物合成的基因。对于开发有效的抗癌药物或药物递送系统，有必要对新的硫酸化多糖的生物活性、构效关系及其生物合成进行进一步研究。
