Farid Wajiha, Masud Tariq, Sohail Asma, Ahmad Nazir, Naqvi S M Saqlan, Khan Sipper, Ali Amjad, Khalifa Salah A, Hussain Abid, Ali Sartaj, Saghir Maryum, Siddeeg Azhari, Manzoor Muhammad Faisal
Department of Food Technology Pir Mehr Ali Shah, Arid Agriculture University Rawalpindi Pakistan.
Institute of Food & Home Sciences Government College University Faisalabad Pakistan.
Food Sci Nutr. 2021 Jul 13;9(9):5092-5102. doi: 10.1002/fsn3.2468. eCollection 2021 Sep.
Strains of WFA1 (KU877440), WFA2 (KU877441), and WFA3 (KU877442) were isolated from indigenous Dahi (yogurt), screened, and selected based on acid and bile tolerance along with the antimicrobial activity. These selected strains were further assessed for their probiotic and functional attributes. Results for simulated gastric and intestinal tolerance/ resistance revealed that all three strains can resist and survive under the following mentioned conditions. To access cell surface hydrophobicity, bacterial adhesion to hydrocarbons (BATH), cellular auto-aggregation, and salt aggregation were performed. In BATH, adhesion of strains against three hydrocarbons namely xylene, dichloromethane, and hexadecane was conducted. The results show that strains showed the least adhesion to xylene (54.25%) as compared to dichloromethane (55.25%) and hexadecane (56.65%). WFA1 showed maximum adherence percentage (55.48%) followed WFA2 (55.48%) and WFA3 (51.38%). Cellular auto-aggregation varied from 21.72% to 30.73% for WFA3 and WFA1, respectively. In the salt aggregation test (SAT), WFA1, WFA2, and WFA3 aggregated at 0.6, 1.0, and 2.0 molar concentrations of ammonium sulfate, respectively. PCR amplification of bile salt hydrolase gene (bsh) was performed and sequences were submitted to the public database of NCBI and Gene bank under accession numbers, KY689139, KY689140, and KY689141. Additionally, a cholesterol-lowering assay was conducted and up to 26% reduction in cholesterol was observed by the strains. Regarding functional properties, exopolysaccharide (EPS) production, and antioxidant potential, strain WFA1 showed promising results EPS (1.027mg/ml), DPPH (80.66%), ABTS (81.97%), and reducing power (1.787). It can be concluded from the present study that the mentioned strains of (WFA1, WFA2, and WFA3) are strongly hydrophobic; thus having an ability to survive and colonize under the gastrointestinal tract which confirms their probiotic nature. Regarding their functional properties, WFA1 (KU877440) showed excellent properties of antioxidants and EPS production.
从本土达希(酸奶)中分离出WFA1(KU877440)、WFA2(KU877441)和WFA3(KU877442)菌株,基于酸耐受性、胆汁耐受性以及抗菌活性进行筛选和选择。对这些选定的菌株进一步评估其益生菌和功能特性。模拟胃和肠道耐受性/抗性的结果表明,所有三种菌株在以下所述条件下都能抵抗并存活。为了评估细胞表面疏水性,进行了细菌对碳氢化合物的粘附(BATH)、细胞自聚集和盐聚集实验。在BATH实验中,检测了菌株对三种碳氢化合物二甲苯、二氯甲烷和十六烷的粘附情况。结果表明,与二氯甲烷(55.25%)和十六烷(56.65%)相比,菌株对二甲苯的粘附最少(54.25%)。WFA1的粘附百分率最高(55.48%),其次是WFA2(55.48%)和WFA3(51.38%)。WFA3和WFA1的细胞自聚集率分别在21.72%至30.73%之间。在盐聚集实验(SAT)中,WFA1、WFA2和WFA3分别在硫酸铵浓度为0.6、1.0和2.0摩尔时发生聚集。进行了胆盐水解酶基因(bsh)的PCR扩增,并将序列提交至NCBI和基因库的公共数据库,登录号分别为KY689139、KY689140和KY689141。此外,进行了降胆固醇实验,观察到这些菌株使胆固醇降低了26%。关于功能特性、胞外多糖(EPS)产生和抗氧化潜力,菌株WFA1表现出了良好的结果,EPS产量为(1.027mg/ml),DPPH清除率为(80.66%),ABTS清除率为(81.97%),还原力为(1.787)。从本研究可以得出结论,上述(WFA1、WFA2和WFA3)菌株具有很强的疏水性,因此有能力在胃肠道中存活和定殖,这证实了它们的益生菌性质。关于它们的功能特性,WFA1(KU877440)表现出优异的抗氧化和EPS产生特性。