Programa de Pós-Graduação em Biodiversidade e Biotecnologia da Rede Bionorte, Universidade do Estado do Amazonas, Escola Superior de Ciências da Saúde, Av. Carvalho Leal, 1777, Manaus, AM, 69065-001, Brazil.
Laboratório de Química Aplicada à Tecnologia, Universidade do Estado do Amazonas, Escola Superior de Ciências da Saúde, Av. Darcy Vargas, 1200, Manaus, AM, 69050-020, Brazil.
Appl Biochem Biotechnol. 2021 Jul;193(7):2145-2161. doi: 10.1007/s12010-021-03542-8. Epub 2021 Mar 5.
Microbial biofilms can cause serious health problems, since, due to their persistent character, they often function as spreaders of contaminants. Hydrolytic enzymes have a number of industrial applications and have been indicated as an alternative to the traditional chemical methods that are used to eradicate microbial biofilms. In this study, we evaluated the ability of enzymatic extracts produced by endophytic fungi isolated from the Amazonian species Myrcia guianensis to remove Staphylococcus aureus biofilms. After culture in liquid medium, the fungal hydrolytic extracts showed amylase (3.77 U/mL), lipase (3.84 U/mL), protease (3.63 U/mL), and xylanase (2.91 U/mL) activity. A 24 h mature S. aureus ATCC6538 biofilm was exposed to each enzyme extract with standardized enzyme activities for 10, 30, and 60 min. The optical density at 630 nm was used to calculate the growth rate (GR%) and the residual biofilm rate (RBR%). The most promising solutions were used in combination, based on a 2 factorial design for 0, 10, 20, and 30 min of exposure. Lipase and protease solutions, when applied separately, were the most effective, and promoted the complete removal of S. aureus biofilms in t (lipase) and t and t (lipase and protease). Of the combined treatments using 1.0 U/mL protease and 0.4 U/mL lipase, total biofilm degradation was observed for all exposure times. Thus, the hydrolases produced by the Amazonian endophytic fungi evaluated here are highlighted as an interesting tool in the fight against microbial biofilms.
微生物生物膜会导致严重的健康问题,因为它们具有持久的特性,往往是污染物传播的媒介。水解酶在许多工业应用中都有应用,并已被证明是替代传统化学方法的一种选择,这些传统化学方法用于消除微生物生物膜。在这项研究中,我们评估了从亚马逊物种杨梅中分离出的内生真菌产生的酶提取物去除金黄色葡萄球菌生物膜的能力。在液体培养基中培养后,真菌水解提取物显示出淀粉酶(3.77 U/mL)、脂肪酶(3.84 U/mL)、蛋白酶(3.63 U/mL)和木聚糖酶(2.91 U/mL)活性。将 24 小时成熟的金黄色葡萄球菌 ATCC6538 生物膜暴露于具有标准化酶活性的每种酶提取物中 10、30 和 60 分钟。使用 630nm 的吸光度来计算生长率(GR%)和残留生物膜率(RBR%)。最有前途的解决方案基于暴露 0、10、20 和 30 分钟的 2 因素设计进行组合,根据 2 因素设计,使用蛋白酶和脂肪酶的单独溶液是最有效的,并且在 t(脂肪酶)和 t 和 t(脂肪酶和蛋白酶)中促进了金黄色葡萄球菌生物膜的完全去除。在使用 1.0 U/mL 蛋白酶和 0.4 U/mL 脂肪酶的联合处理中,观察到所有暴露时间的总生物膜降解。因此,这里评估的亚马逊内生真菌产生的水解酶被强调为对抗微生物生物膜的一种有趣工具。