Nirwati Hera, Damayanti Ema, Sholikhah Eti Nurwening, Mutofa Mustofa, Widada Jaka
Department of Microbiology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia.
Research Center for Food Technology and Processing, National Research and Innovation Agency, Gunungkidul, 55861, Indonesia.
Heliyon. 2022 Apr 25;8(4):e09333. doi: 10.1016/j.heliyon.2022.e09333. eCollection 2022 Apr.
Biofilm-forming fungi, , are currently a serious problem in infectious disease cases. Soil bacteria sp. GMR22 have a large genome size and antifungal metabolites against . , but its potential antibiofilm activity is not clearly defined. The aims of this study were to determine the antibiofilm activity of GMR22 against , identify the main constituents of active extracts, and investigate the biosynthesis gene clusters encoding the enzymes related to metabolism pathways. Antifungal and antibiofilm measurements were performed using in vitro assays on ATCC 10231. Main constituents of active extracts were analyzed using untargeted Liquid Chromatography tandem High-Resolution Mass Spectrometry (LC-HRMS). RAST software was applied to investigate the gene clusters of the biosynthesis pathways based on whole genome sequences. Chloroform extract of GMR22 has antifungal and antibiofilm properties at 13-420 μg/mL with palmitic acid (CHO, 273.27028 Da), a saturated fatty acid as a major constituent (42.74). sp. GMR22 has 53 subsystems related to fatty acids biosynthesis (Fab) FAS II. The Kyoto Encyclopedia of Gene and Genome map of Fab revealed 10 of 21 (47.6%) gene clusters encode enzymes related to Fab. There were six gene clusters encoding the enzymes related to the hexadecenoic acid (palmitic acid) biosynthesis pathways: 6.4.12; FabD, FabH, FabF, FabG, FabI and 1.14.192. Each enzyme was encoded by 3-14 genes. These results confirmed that soil sp. GMR22 bacterium has remarkable biotechnological potential by producing fatty acids which are mostly palmitic acid as an active antibiofilm agent against .
形成生物膜的真菌，目前在传染病病例中是一个严重问题。土壤细菌GMR22菌株具有较大的基因组大小，且含有针对[真菌名称未给出]的抗真菌代谢产物。然而，其潜在的抗生物膜活性尚未明确界定。本研究的目的是确定GMR22对[真菌名称未给出]的抗生物膜活性，鉴定活性提取物的主要成分，并研究编码与代谢途径相关酶的生物合成基因簇。使用体外试验对[真菌名称未给出]ATCC 10231进行抗真菌和抗生物膜测量。利用非靶向液相色谱串联高分辨率质谱（LC-HRMS）分析活性提取物的主要成分。基于全基因组序列，应用RAST软件研究生物合成途径的基因簇。GMR22的氯仿提取物在13 - 420μg/mL时具有抗真菌和抗生物膜特性，主要成分是棕榈酸（CHO，273.27028 Da），一种饱和脂肪酸（占42.74%）。GMR22菌株有53个与脂肪酸生物合成（Fab）FAS II相关的子系统。Fab的京都基因与基因组百科全书图谱显示，21个基因簇中有10个（47.6%）编码与Fab相关的酶。有6个基因簇编码与十六碳烯酸（棕榈酸）生物合成途径相关的酶：6.4.12；FabD、FabH、FabF、FabG、FabI和1.14.192。每种酶由3 - 14个基因编码。这些结果证实，土壤GMR22菌株通过产生主要为棕榈酸的脂肪酸作为针对[真菌名称未给出]的活性抗生物膜剂，具有显著的生物技术潜力。
