Rodrigues Josy Caldas, Lima da Silva Weison, Ribeiro da Silva David, Maia Carolina Rabelo, Santos Goiabeira Clarice Virginia, Figueiredo Chagas Haile Dean, Ayres D'Elia Gigliola Mayara, Barbosa Alves Gleica Soyan, Zahner Viviane, Nunez Cecilia Veronica, Cristo Fernandes Ormezinda Celeste
Leônidas and Maria Deane Institute-ILMD/Fiocruz Amazônia, Terezina street, 476, Adrianópolis, Manaus, Brazil.
National Institute of Amazonian Research, André Araujo Avenue, 2936, Petrópolis, Manaus, Brazil.
Int J Microbiol. 2022 May 17;2022:4010018. doi: 10.1155/2022/4010018. eCollection 2022.
The antimicrobial potential of sp., isolated from the Amazon biome, which is stored at the Amazon Fungi Collection-CFAM at ILMD/FIOCRUZ, was evaluated. The fungal culture was cultivated in yeast extract agar and sucrose (YES) for cold extraction of the biocompounds in ethyl acetate at 28 °C for 7 days in a BOD type incubator. The obtained extract was evaluated for its antimicrobial activity against and Gram-positive and negative bacteria by the "cup plate" method and the determination of the minimum inhibitory concentration (MIC) by the broth microdilution method. The extract was subjected to thin layer chromatography (TLC) and fractionated by open and semipreparative column chromatography. The fractions of interest had their chemical constituents elucidated by nuclear magnetic resonance and mass spectrometry. The elucidated molecule was evaluated for cytotoxicity against the human fibroblast strain (MRC5). The extract presented inhibitory activity against both Gram-positive and negative bacteria, with the range of inhibition halos from 5.3 to 14 mm in diameter and an MIC ranging from 500 to 15.6 g/mL. Seventy-one fractions were collected and TLC analysis suggested the presence of substances with double bond groups: coumarins, flavonoids, phenolic, alkaloids, and terpenes. NMR and MS analyses demonstrated that the isolated molecule was kojic acid. The results of the cytotoxicity test showed that MRC5 cells presented viability at concentrations from 500 to 7.81 g/mL. The kojic acid molecule of sp., with antibacterial activity and moderate toxicity at the concentrations tested, is a promising prototype of an alternative active principle of an antimicrobial drug.
对从亚马逊生物群落分离出的、保存在ILMD/FIOCRUZ的亚马逊真菌收藏中心（CFAM）的某菌株的抗菌潜力进行了评估。将真菌培养物在酵母提取物琼脂和蔗糖（YES）培养基中培养，于28°C在BOD型培养箱中放置7天，以便用乙酸乙酯对生物化合物进行冷萃取。通过“杯碟法”评估所得提取物对革兰氏阳性和阴性细菌的抗菌活性，并通过肉汤微量稀释法测定最低抑菌浓度（MIC）。对提取物进行薄层色谱（TLC）分析，并通过开放柱色谱和半制备柱色谱进行分离。通过核磁共振和质谱对感兴趣的馏分的化学成分进行阐明。对阐明的分子进行针对人成纤维细胞系（MRC5）的细胞毒性评估。该提取物对革兰氏阳性和阴性细菌均表现出抑制活性，抑制圈直径范围为5.3至14毫米，MIC范围为500至15.6微克/毫升。收集了71个馏分，TLC分析表明存在含双键基团的物质：香豆素、黄酮类、酚类、生物碱和萜类。核磁共振和质谱分析表明分离出的分子是曲酸。细胞毒性测试结果表明，MRC5细胞在浓度为500至7.81微克/毫升时具有活力。在测试浓度下具有抗菌活性和中等毒性的某菌株的曲酸分子，是一种有前景的抗菌药物替代活性成分原型。
