Zhang Fan, Yang Hongjian, Wang Wei, Cao Zhijun, Li Shengli, Wang Weikang
State Key Laboratory of Animal Nutrition and Feeding, College of Animal Science and Technology, China Agricultural University, Beijing, 100193, P.R. China.
Probiotics Antimicrob Proteins. 2025 Jul 26. doi: 10.1007/s12602-025-10673-5.
Probiotics identification requires comprehensive safety assessment and probiotic property evaluation. In this study, a Pediococcus acidilactici PA_ZFRA1 strain isolated from the bovine rumen was first characterized by whole-genome sequencing and probiotic properties. The ZFRA1 genome is 2,061,765 bp in length, with 1982 predicted coding sequences and an average GC content of 42.26%. For safety assessment: ZFRA1 shows no hemolytic activity, lacks decarboxylase genes related to biogenic amine synthesis, and poses negligible risk of horizontal transfer for antibiotic resistance genes and virulence factor genes. Regarding probiotic properties, the stress resistance assays demonstrated that ZFRA1 can withstand acid (pH 2, 3, and 4), simulated gastrointestinal fluids, and bile salts (0.3% and 1%). Furthermore, ZFRA1 displayed a considerable antioxidant capacity (75.60% DPPH, 77.35% ABTS free radical scavenging rates, and 1.56 mM TEAC total antioxidant capacity), moderate hydrophobicity (10.42%-20.83%), an auto-aggregation rate of 70.79% at 24 h, an adhesion rate to Caco-2 cells of 11.06%, and high co-aggregation ability (76.50%-83.66%). Moreover, ZFRA1 exhibited inhibitory ability against seven pathogens (E. coli CICC 10372, S. aureus ATCC 27217, C. perfringens ATCC 13124, B. cereus CICC 21290, P. aeruginosa ATCC 10145, S. typhimurium ATCC 14028, and L. monocytogenes ATCC 19111). In summary, the results of genomic analysis combined with in vitro assays showed that ZFRA1 is a promising candidate probiotic strain. However, further in vivo studies are required to validate its safety and efficacy before it can be considered for food fermentation, human consumption, or animal nutrition applications.
益生菌的鉴定需要全面的安全性评估和益生菌特性评价。在本研究中,首先通过全基因组测序和益生菌特性对从牛瘤胃中分离出的一株嗜酸乳杆菌PA_ZFRA1进行了表征。ZFRA1基因组长度为2,061,765 bp,有1982个预测编码序列,平均GC含量为42.26%。安全性评估方面:ZFRA1无溶血活性,缺乏与生物胺合成相关的脱羧酶基因,抗生素抗性基因和毒力因子基因的水平转移风险可忽略不计。关于益生菌特性,抗逆性试验表明ZFRA1能耐受酸(pH 2、3和4)、模拟胃肠液和胆盐(0.3%和1%)。此外,ZFRA1表现出相当的抗氧化能力(DPPH清除率75.60%、ABTS自由基清除率77.35%、总抗氧化能力1.56 mM TEAC)、适度的疏水性(10.42%-20.83%)、24小时自聚集率70.79%、对Caco-2细胞的黏附率11.06%以及高共聚集能力(76.50%-83.66%)。此外,ZFRA1对七种病原菌(大肠杆菌CICC 10372、金黄色葡萄球菌ATCC 27217、产气荚膜梭菌ATCC 13124、蜡样芽孢杆菌CICC 21290、铜绿假单胞菌ATCC 10145、鼠伤寒沙门氏菌ATCC 14028和单核细胞增生李斯特菌ATCC 19111)具有抑制能力。总之,基因组分析结果与体外试验表明ZFRA1是一种有前景的益生菌候选菌株。然而,在将其考虑用于食品发酵、人类消费或动物营养应用之前,还需要进一步的体内研究来验证其安全性和有效性。