Wu Ruiyun, Qin Yuxuan, Shen Qian, Li Pinglan
1Key Laboratory of Precision Nutrition and Food Quality, Key Laboratory of Functional Dairy, Ministry of Education; College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qinghua East Road, East Campus, Haidian District, Beijing, 100083 China.
3Department of Biology, Northeastern University, Boston, MA 02115 USA.
3 Biotech. 2020 Jun;10(6):243. doi: 10.1007/s13205-020-02228-y. Epub 2020 May 9.
LPL061, which shows strong exopolysaccharide (EPS) producing capacity, was isolated from carnations in Beijing, China. The complete genome of LPL061 comprised a single circular chromosome (3,907,268 bp; G+C content of 46.7%) with 3,737 coding DNA sequences, 26 rRNA, and 89 tRNA. According to genome analysis, 12 protein-coding genes which related to polysaccharide biosynthesis in LPL061 were identified. Comparative genome analysis revealed that the EPS biosynthetic gene cluster was relatively conserved among species. EPS showed approximately 60% inhibitory activity on the α-glucosidase at 100 μg/mL. The results of quantitative reverse transcription PCR further demonstrated that compared to insulin-resistant model with insulin (500 μg/mL) (without EPS treatment), the insulin-resistant HepG2 cells treated with EPS decreased the expression of phosphoenolpyruvate carboxykinase () from 4.425 to 0.1587, glucose-6-phosphatase () decreased from 4.272 to 0.1929, and glycogen synthase kinase3β (()) decreased from 2.451 to 0.993, respectively. Meanwhile, EPS treatment increased GS expression and resulted in intracellular glycogen concentration increased from 28.30% to 86.48%, which further supported that EPS form LPL061 could reduce the concentration of blood glucose effectively. These results could be beneficial for better understanding of the hypoglycemic mechanism of LPL061 EPS and developing an EPS-based anti-diabetic agent in the future.
LPL061是从中国北京的康乃馨中分离得到的,具有很强的胞外多糖(EPS)产生能力。LPL061的完整基因组由一条单一的环状染色体(3,907,268 bp;G+C含量为46.7%)组成,包含3,737个编码DNA序列、26个rRNA和89个tRNA。根据基因组分析,在LPL061中鉴定出12个与多糖生物合成相关的蛋白质编码基因。比较基因组分析表明,EPS生物合成基因簇在不同物种间相对保守。EPS在100 μg/mL时对α-葡萄糖苷酶显示出约60%的抑制活性。定量逆转录PCR结果进一步表明,与用胰岛素(500 μg/mL)处理的胰岛素抵抗模型(未用EPS处理)相比,用EPS处理的胰岛素抵抗HepG2细胞中,磷酸烯醇式丙酮酸羧激酶()的表达从4.425降至0.1587,葡萄糖-6-磷酸酶()从4.272降至0.1929,糖原合酶激酶3β()从2.451降至0.993。同时,EPS处理增加了GS的表达,并使细胞内糖原浓度从28.30%增加到86.48%,这进一步证明来自LPL061的EPS可以有效降低血糖浓度。这些结果有助于更好地理解LPL061 EPS的降血糖机制,并为未来开发基于EPS的抗糖尿病药物提供帮助。