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从发酵的罗伊氏乳杆菌中筛选具有降血糖特性的潜在新型益生菌RAMULAB52

Screening for potential novel probiotic RAMULAB52 with antihyperglycemic property from fermented L.

作者信息

Sreepathi Navya, Kumari V B Chandana, Huligere Sujay S, Al-Odayni Abdel-Basit, Lasehinde Victor, Jayanthi M K, Ramu Ramith

机构信息

Department of Biotechnology and Bioinformatics, JSS Academy of Higher Education and Research, Mysuru, Karnataka, India.

Department of Pharmacology, JSS Medical College, JSS Academy of Higher Education and Research, Mysuru, Karnataka, India.

出版信息

Front Microbiol. 2023 Jun 20;14:1168102. doi: 10.3389/fmicb.2023.1168102. eCollection 2023.

Abstract

Probiotics are live microorganisms with various health benefits when consumed in appropriate amounts. Fermented foods are a rich source of these beneficial organisms. This study aimed to investigate the probiotic potential of lactic acid bacteria (LAB) isolated from fermented papaya ( L.) through methods. The LAB strains were thoroughly characterized, considering their morphological, physiological, fermentative, biochemical, and molecular properties. The LAB strain's adherence and resistance to gastrointestinal conditions, as well as its antibacterial and antioxidant capabilities, were examined. Moreover, the strains were tested for susceptibility against specific antibiotics, and safety evaluations encompassed the hemolytic assay and DNase activity. The supernatant of the LAB isolate underwent organic acid profiling (LCMS). The primary objective of this study was to assess the inhibitory activity of α-amylase and α-glucosidase enzymes, both and . Gram-positive strains that were catalase-negative and carbohydrate fermenting were selected for further analysis. The LAB isolate exhibited resistance to acid bile (0.3% and 1%), phenol (0.1% and 0.4%), and simulated gastrointestinal juice (pH 3-8). It demonstrated potent antibacterial and antioxidant abilities and resistance to kanamycin, vancomycin, and methicillin. The LAB strain showed autoaggregation (83%) and adhesion to chicken crop epithelial cells, buccal epithelial cells, and HT-29 cells. Safety assessments indicated no evidence of hemolysis or DNA degradation, confirming the safety of the LAB isolates. The isolate's identity was confirmed using the 16S rRNA sequence. The LAB strain RAMULAB52, derived from fermented papaya, exhibited promising probiotic properties. Moreover, the isolate demonstrated significant inhibition of α-amylase (86.97%) and α-glucosidase (75.87%) enzymes. studies uncovered that hydroxycitric acid, one of the organic acids derived from the isolate, interacted with crucial amino acid residues of the target enzymes. Specifically, hydroxycitric acid formed hydrogen bonds with key amino acid residues, such as GLU233 and ASP197 in α-amylase, and ASN241, ARG312, GLU304, SER308, HIS279, PRO309, and PHE311 in α-glucosidase. In conclusion, RAMULAB52, isolated from fermented papaya, possesses promising probiotic properties and exhibits potential as an effective remedy for diabetes. Its resistance to gastrointestinal conditions, antibacterial and antioxidant abilities, adhesion to different cell types, and significant inhibition of target enzymes make it a valuable candidate for further research and potential application in the field of probiotics and diabetes management.

摘要

益生菌是适量食用时对健康有多种益处的活微生物。发酵食品是这些有益微生物的丰富来源。本研究旨在通过多种方法研究从发酵木瓜中分离出的乳酸菌(LAB)的益生菌潜力。考虑到乳酸菌菌株的形态、生理、发酵、生化和分子特性,对其进行了全面表征。检测了乳酸菌菌株对胃肠道条件的粘附性和抗性,以及其抗菌和抗氧化能力。此外,还测试了这些菌株对特定抗生素的敏感性,安全评估包括溶血试验和DNase活性。对乳酸菌分离株的上清液进行了有机酸分析(LCMS)。本研究的主要目的是评估α-淀粉酶和α-葡萄糖苷酶的抑制活性,包括体外和体内。选择过氧化氢酶阴性且能发酵碳水化合物的革兰氏阳性菌株进行进一步分析。乳酸菌分离株对酸性胆汁(0.3%和1%)、苯酚(0.1%和0.4%)和模拟胃肠液(pH 3 - 8)具有抗性。它表现出强大的抗菌和抗氧化能力,以及对卡那霉素、万古霉素和甲氧西林的抗性。乳酸菌菌株表现出自动聚集(83%),并能粘附于鸡嗉囊上皮细胞、颊上皮细胞和HT - 29细胞。安全评估表明没有溶血或DNA降解的迹象,证实了乳酸菌分离株的安全性。使用16S rRNA序列确认了分离株的身份。源自发酵木瓜的乳酸菌菌株RAMULAB52具有良好的益生菌特性。此外,该分离株对α-淀粉酶(86.97%)和α-葡萄糖苷酶(75.87%)表现出显著抑制作用。进一步研究发现,该分离株产生的有机酸之一羟基柠檬酸与目标酶的关键氨基酸残基相互作用。具体而言,羟基柠檬酸与关键氨基酸残基形成氢键,如α-淀粉酶中的GLU233和ASP197,以及α-葡萄糖苷酶中的ASN241、ARG312、GLU304、SER308、HIS279、PRO309和PHE311。总之,从发酵木瓜中分离出的RAMULAB52具有良好的益生菌特性,作为糖尿病的有效治疗方法具有潜力。其对胃肠道条件的抗性、抗菌和抗氧化能力、对不同细胞类型的粘附性以及对目标酶的显著抑制作用使其成为益生菌和糖尿病管理领域进一步研究和潜在应用的有价值候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c3d0/10318367/e624cc83cf7e/fmicb-14-1168102-g0001.jpg

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