| 梁美婷,杨珊珊,贺 怡,玛依娜·卡哈尔,陈邬锦,李 瑞,孙玉萍.一株人肠源性高效尿酸降解菌的分离、鉴定及降尿酸条件优化[J].现代生物医学进展英文版,2023,(21):4013-4019. |
| 一株人肠源性高效尿酸降解菌的分离、鉴定及降尿酸条件优化 |
| Isolation, Identification and Optimization of Uric Acid Degradation Conditions of a Human Enterogenic High Efficiency Uric Acid Degrading Bacterium |
| Received:March 20, 2023 Revised:April 14, 2023 |
| DOI：10.13241/j.cnki.pmb.2023.21.003 |
| 中文关键词: 肠道微生物 尿酸降解菌 尿酸降解率 |
| 英文关键词: Gut microbiome Uric acid degrading bacteria Uric acid degradation rate |
| 基金项目:国家自然科学基金项目（82260182；81960169）；新疆维吾尔自治区自然科学基金项目（2021D01C275；2022D01C185）；新疆维吾尔自治区名医名方与特色重点实验室开放课题（2020D04024）；新疆维吾尔自治区高校科研计划项目（XJEDU2021Y054）；新疆研究生科研创新项目（XJ2021G228；XJ2022G166）；克拉玛依市创新人才项目（20212022hjcxrc0060） |
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| 中文摘要: |
| 摘要 目的：肠道作为人体的重要消化器官，其内定植的微生物在尿酸合成和代谢过程中发挥着重要作用，本研究利用含尿酸靶向培养基筛选正常人群肠道内具有降尿酸功能的细菌并鉴定。方法：依据尿酸的摩尔质量制备含不同浓度尿酸的BHI培养基，液体培养基扩增并驯化肠道粪便微生物，固体培养基分离和纯化具有尿酸降解功能的细菌。挑取固体培养基上形态一致的单个菌落进行革兰氏染色和镜检，筛选出已纯化菌株，在需氧和厌氧培养条件下测定尿酸降解率，选取降解率≥50%以上的菌株为高效尿酸降解菌的候选菌株，再测定不同温度和pH值条件下的尿酸降解率，进行降尿酸条件优化。利用16S rDNA序列测定法对尿酸降解菌进行鉴定，药敏实验测定该菌对抗生素的敏感性。结果：正常人群粪便微生物中分离获得一株高效尿酸降解菌B5C，第5天需氧条件下的尿酸降解率均>50%，与初始尿酸浓度相比具有统计学意义（P＜0.05）。优化降尿酸条件后，在37℃、pH7.0时，降解率可达88.7%，经鉴定为粪肠球菌，对常见的抗生素如阿莫西林、氨苄西林和青霉素G等具有较高的敏感性。结论：本研究利用含不同尿酸浓度的靶向培养基驯化、分离和鉴定出一株人肠源性细菌，在需氧条件下也具有较高的尿酸降解率，可为今后临床降尿酸微生物制剂的开发和利用提供新的菌种资源。 |
| 英文摘要: |
| ABSTRACT Objective: As an important digestive organ in the human body, the microbiome colonized in the intestine play an important role in the synthesis and metabolism of uric acid. In this study, we used uric acid-containing targeting medium to screen for and identify bacteria with uric acid degrading function in the normal human intestine. Methods: Preparation of BHI medium containing different concentrations of uric acid based on the molar mass of uric acid. Liquid medium to amplify and domesticate intestinal bacteria, solid medium to isolate and purify bacteria with uric acid degradation function. Single colonies with consistent morphology on solid medium were picked for Gram staining and microscopic examination. The purified strains were screened and the rate of uric acid degradation was determined under aerobic and anaerobic culture conditions. The strains with degradation rate ≥50% or more were selected as the candidate strains of efficient uric acid-degrading bacteria. Subsequently, the degradation rate of uric acid was measured under different temperature and pH conditions for optimization of uric acid reduction conditions. The 16S rDNA sequence assay was used to identify the uric acid-degrading bacteria, and drug sensitivity test to determine the susceptibility of the bacteria to antibiotics. Results: An efficient strain of uric acid-degrading bacteria B5C was isolated from fecal microbiome of normal people, showed >50% degradation of uric acid on day 5 under aerobic conditions, which was statistically significant compared to the initial uric acid concentration (P<0.05). After optimizing the conditions for degradation of uric acid, the degradation rate could reach 88.7% at 37℃ and pH 7.0. It was identified as Enterococcus faecalis, which has high sensitivity to common antibiotics such as amoxicillin, ampicillin and penicillin G. Conclusion: In this study, a strain of human enteric-derived bacteria was domesticated, isolated and characterized using targeted media containing different uric acid concentrations. It also has a high rate of uric acid degradation under aerobic conditions, providing a new strain resource for the development and utilization of uric acid degrading microbial preparations. |
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