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从淡红链霉菌ICN3中制备抗耐甲氧西林金黄色葡萄球菌(MRSA)的化合物及其在斑马鱼胚胎中的评估。

Production of a compound against methicillin resistant Staphylococcus aureus (MRSA) from Streptomyces rubrolavendulae ICN3 & its evaluation in zebrafish embryos.

作者信息

Kannan Rajaretinam Rajesh, Iniyan Appadurai Muthamil, Vincent Samuel Gnana Prakash

机构信息

International Centre for Nanobiotechnology, Centre for Marine Science & Technology, Manonmaniam Sundaranar University, Rajakkamangalam, India.

出版信息

Indian J Med Res. 2014 Jun;139(6):913-20.

Abstract

BACKGROUND & OBJECTIVES: Antibiotic resistance in pathogens has become a serious problem worldwide. Therefore, the search for new antibiotics for drug resistanct pathogens is an important endeavor. The present study deals with the production of anti-methicillin resistant Staphylococcus aureus (MRSA) potential of Streptomyces rubrolavendulae ICN3 and evaluation of anti-MRSA compound in zebrafish embryos.

METHODS

The antibiotic production from S. rubrolavendulae ICN3 was optimized in solid state fermentation and extracted. The antagonistic activity was confirmed against MRSA and purified in silica gel column and reverse phase--HPLC with an absorption maximum at 215 nm. Minimal inhibitory concentration of the compound was determined by broth microdilution method. Zebrafish embryos were used to evaluate the extract/compound for its minimal inhibition studies, influences on heart beat rates, haematopoietic blood cell count and lethal dose values.

RESULTS

Streptomyces rubrolavendulae ICN3 showed potent antagonistic activity against MRSA with a zone of 42 mm. The minimum inhibitory concentration was calculated as 500 μg/ml of the crude extract and the purified C23 exhibited 2.5 μg/ml in in vitro assay. The LC 50 value of the anti MRSA compound C23 was calculated as 60.49 μg/ml and the MRSA treated embryos survived in the presence of purified compound C23 at a dose of 10 μg/ml.

INTERPRETATION & CONCLUSIONS: Our results suggested that the compound was potent with less toxic effects in zebrafish embryonic model system for MRSA infection. Further structural evaluation and analysis in higher mammalian model system may lead to a novel drug candidate for drug resistant Staphylococcus aureus.

摘要

背景与目的

病原体中的抗生素耐药性已成为全球范围内的一个严重问题。因此,寻找针对耐药病原体的新型抗生素是一项重要的工作。本研究涉及淡紫红色链霉菌ICN3抗耐甲氧西林金黄色葡萄球菌(MRSA)潜力的产生以及斑马鱼胚胎中抗MRSA化合物的评估。

方法

在固态发酵中优化淡紫红色链霉菌ICN3的抗生素生产并进行提取。确认其对MRSA的拮抗活性,并在硅胶柱和反相高效液相色谱中进行纯化,最大吸收波长为215nm。通过肉汤微量稀释法测定该化合物的最低抑菌浓度。使用斑马鱼胚胎评估提取物/化合物的最低抑制研究、对心率、造血血细胞计数和致死剂量值的影响。

结果

淡紫红色链霉菌ICN3对MRSA表现出强大的拮抗活性,抑菌圈为42mm。粗提物的最低抑菌浓度计算为500μg/ml,纯化后的C23在体外试验中表现出2.5μg/ml。抗MRSA化合物C23的LC50值计算为60.49μg/ml,在10μg/ml剂量的纯化化合物C23存在下,经MRSA处理的胚胎存活。

解读与结论

我们的结果表明,该化合物在斑马鱼胚胎模型系统中对MRSA感染有效且毒性较小。在更高等的哺乳动物模型系统中进行进一步的结构评估和分析可能会产生一种针对耐甲氧西林金黄色葡萄球菌的新型候选药物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48e5/4165004/ca4b74c96086/IJMR-139-913-g001.jpg

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