de Souza Freitas Fernanda, Coelho de Assis Lage Tiago, Ayupe Bruna Almeida Leão, de Paula Siqueira Tatiane, de Barros Mariana, Tótola Marcos Rogério
Laboratório de Biotecnologia e Biodiversidade Para o Meio Ambiente, Departamento de Microbiologia, Universidade Federal de Viçosa, Peter Henry Rolfs Avenue w/n, Viçosa, Minas Gerais Brazil.
Laboratório de Doenças Bacterianas, Departamento de Veterinária, Universidade Federal de Viçosa, Peter Henry Rolfs Avenue w/n, Viçosa, Minas Gerais Brazil.
3 Biotech. 2020 Nov;10(11):474. doi: 10.1007/s13205-020-02459-z. Epub 2020 Oct 13.
This study aimed to investigate the antimicrobial, antibiofilm, and cytotoxic effects of biosurfactant lipopeptides synthesized by TR47II. For this purpose, the lipopeptides were partially purified using a three-step process and characterized. In the first step, the crude extract obtained from acid precipitation exhibited strong antibacterial activity against the Gram-negative opportunistic pathogens ATCC 8750, ATCC 13138, ATCC 14909, and ATCC 15175. Moreover, partial inhibition was observed against ATCC 13048 (42%), ATCC 25922 (16%), and ATCC 27853 (47%). The lipopeptides in the crude extract were extracted with methanol and fractioned on a silica gel chromatography column, rendering four TLC-pooled chromatographic fractions, named F1, F2, F3, and F4. The chromatographic fraction F4 was the most bioactive, with MIC values between 300 and 600 µg mL. Besides, F4 at sub-MIC doses dislodged the biofilms of , , and by about 100, 85, and 81%, respectively. No cytotoxic effect was observed in mammalian cells at MIC. MALDI-TOF-MS analysis revealed that F4 contained cyclic lipopeptides belonging to two families: iturins (m/z 1004 to 1087) and fengycins (m/z 1424 to 1545). The dual effect of F4 on planktonic and sessile growth could suggest that the synergistic application of these biosurfactants could be efficient in the control of these opportunistic pathogens.
本研究旨在调查TR47II合成的生物表面活性剂脂肽的抗菌、抗生物膜和细胞毒性作用。为此,采用三步法对脂肽进行部分纯化并进行表征。第一步,酸沉淀得到的粗提物对革兰氏阴性机会致病菌ATCC 8750、ATCC 13138、ATCC 14909和ATCC 15175表现出强大的抗菌活性。此外,还观察到对ATCC 13048(42%)、ATCC 25922(16%)和ATCC 27853(47%)有部分抑制作用。粗提物中的脂肽用甲醇萃取,并在硅胶色谱柱上进行分离,得到四个薄层层析合并的色谱馏分,命名为F1、F2、F3和F4。色谱馏分F4生物活性最强,MIC值在300至600µg/mL之间。此外,亚MIC剂量的F4分别使 、 和 的生物膜脱落约100%、85%和81%。在MIC浓度下,未在哺乳动物细胞中观察到细胞毒性作用。基质辅助激光解吸电离飞行时间质谱分析表明,F4含有属于两个家族的环状脂肽:iturins(m/z 1004至1087)和fengycins(m/z 1424至1545)。F4对浮游生长和固着生长的双重作用表明,这些生物表面活性剂的协同应用可能对控制这些机会致病菌有效。