Nabila Tsania Taskia, Damayanti Ema, Widada Jaka
Study Program of Biotechnology, Graduate School of Universitas Gadjah Mada (UGM), Jl. Teknika Utara, Pogung, Mlati, Sleman, Yogyakarta 55281, Indonesia; Research Assistant of Food Microbiology Research Group, Research Center for Food Processing and Technology, National Research and Innovation Agency (BRIN), Jl. Jogja Wonosari KM 31.5, Gading, Playen, Gunungkidul 55861, Indonesia.
Research Center for Food Processing and Technology, BRIN, Jl. Jogja Wonosari KM 31.5, Gading, Playen, Gunungkidul 55861, Indonesia.
J Food Prot. 2025 Jun 23;88(7):100546. doi: 10.1016/j.jfp.2025.100546. Epub 2025 May 20.
Food contamination by pathogenic microorganisms has become a significant issue. This study investigated the antibacterial compounds of a Bacillus isolate from stingless bee bread, Bacillus velezensis BP1, against foodborne pathogens Staphylococcus aureus ATCC 25923 and Salmonella enterica Typhimurium ATCC 14028 using the one strain many compounds (OSMAC), metabolomic, and genomic approaches. The culture media used were Tryptic Soy Broth (TSB), TSB with Chitosan (TSB-Chi), several synthetic broths composed of Mineral Salts with Glucose (MG) and Glucose-Fructose (MGF), Carboxymethyl Cellulose (CMC), and Starch Nitrate (SNB), to analyze diverse antibacterial compounds. Twelve extracts from the supernatant (S) and pellet (P) were screened using the microdilution method. P-TSB and S-TSB-Chi demonstrated the highest antibacterial effects, with inhibitory concentration (IC) values of 253.5 ppm and 740.28 ppm, respectively. Fourier transform infrared spectroscopy (FTIR) analysis identified an amide I group contributing to extract clustering. Untargeted liquid chromatography-high resolution mass spectrometry (LC-HRMS) revealed six compounds significantly contributing to extract clustering. Enrichment analysis showed that chitosan was associated with the metabolic processes of pyrimidine and nucleotide metabolisms. Growth curve assay and scanning electron microscopy (SEM) confirmed the extracts' efficacy. The Bacillus isolate showed an average nucleotide identity (ANI) of 98.08% to Bacillus velezensis NRRL B-41580. Genome mining revealed twelve biosynthetic gene clusters, six 100% similar to known clusters. Molecular docking demonstrated that genome mining-derived bacillibactin and LC-HRMS-derived bis(4-ethylbenzylidene)sorbitol and cyclo(phenylalanyl-prolyl) exhibited the strongest binding affinities against four pathogen-associated proteins, outperforming ampicillin. This study highlights Bacillus velezensis BP1's potential as a source of diverse antibacterial compounds.
病原微生物对食品的污染已成为一个重大问题。本研究采用一菌多化合物（OSMAC）、代谢组学和基因组学方法，研究了从无刺蜂面包中分离出的一种芽孢杆菌——贝莱斯芽孢杆菌BP1，对食源性病原体金黄色葡萄球菌ATCC 25923和鼠伤寒沙门氏菌ATCC 14028的抗菌化合物。使用的培养基有胰蛋白胨大豆肉汤（TSB）、添加壳聚糖的TSB（TSB-Chi）、几种由矿物质盐与葡萄糖（MG）和葡萄糖-果糖（MGF）组成的合成肉汤、羧甲基纤维素（CMC）和淀粉硝酸盐（SNB），以分析不同的抗菌化合物。使用微量稀释法对上清液（S）和沉淀（P）的12种提取物进行了筛选。P-TSB和S-TSB-Chi表现出最高的抗菌效果，抑制浓度（IC）值分别为253.5 ppm和740.28 ppm。傅里叶变换红外光谱（FTIR）分析确定了一个酰胺I基团有助于提取物聚类。非靶向液相色谱-高分辨率质谱（LC-HRMS）显示有六种化合物对提取物聚类有显著贡献。富集分析表明壳聚糖与嘧啶和核苷酸代谢的代谢过程有关。生长曲线测定和扫描电子显微镜（SEM）证实了提取物的功效。该芽孢杆菌分离株与贝莱斯芽孢杆菌NRRL B-41580的平均核苷酸同一性（ANI）为98.08%。基因组挖掘揭示了12个生物合成基因簇，其中6个与已知簇100%相似。分子对接表明，基因组挖掘得到的杆菌肽和LC-HRMS得到的双（4-乙基苄叉）山梨醇和环（苯丙氨酰-脯氨酰）对四种病原体相关蛋白表现出最强的结合亲和力，优于氨苄青霉素。本研究突出了贝莱斯芽孢杆菌BP1作为多种抗菌化合物来源的潜力。
