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传统发酵食品中乳酸菌的分子鉴定及利用食品废弃物筛选胞外多糖的产生

Molecular identification of lactic acid bacteria from traditional fermented foods and screening exopolysaccharide production by using food wastes.

作者信息

Karaman Kevser, Sirke Sibel Turan, Rifaioglu Şeyda Nur Türkay

机构信息

Department of Agricultural Biotechnology, Faculty of Agriculture, Erciyes University, Kayseri, Türkiye.

Genome and Stem Cell Research Center, Erciyes University, Kayseri, Türkiye.

出版信息

Folia Microbiol (Praha). 2025 Apr;70(2):391-401. doi: 10.1007/s12223-024-01187-8. Epub 2024 Aug 27.

Abstract

In this study, lactic acid bacteria (LAB) isolation from fermented foods and molecular identification using magnetic bead technology were performed. And then exopolysaccharide (EPS) production possibility was tested in agar medium, and the positive ones were selected for the next step. The bacteria that could produce higher carbohydrate level were grown in MRS medium fortified with whey and pumpkin waste. In our study, 19 different LAB species were identified from fermented products collected from different places in Hatay (Türkiye) province. In molecular identification, universal primer pairs, p806R/p8FPL, and PEU7/DG74 were used for PCR amplification. After that, PCR products purified using paramagnetic bead technology were sequenced by the Sanger sequencing method. The dominant species, 23.8% of the isolates, were identified as Lactiplantibacillus plantarum. As a technological property of LAB, exopolysaccharide production capability of forty-two LAB isolate was tested in agar medium, and after eleven isolates were selected as positive. Two LAB (Latilactobacillus curvatus SHA2-3B and Loigolactobacillus coryniformis SHA6-3B) had higher EPS production capability when they were grown in MRS broth fortified with pumpkin waste and whey. The highest EPS content (1750 mg/L glucose equivalent) was determined in Loigolactobacillus coryniformis SHA6-3B grown in MRS broth fortified with 10% pumpkin waste. Besides the produced EPS samples were validated with FTIR and SEM methods.

摘要

本研究进行了从发酵食品中分离乳酸菌(LAB)并使用磁珠技术进行分子鉴定。然后在琼脂培养基中测试了胞外多糖(EPS)的产生可能性,并选择阳性菌株进行下一步研究。能产生较高碳水化合物水平的细菌在添加了乳清和南瓜废料的MRS培养基中培养。在我们的研究中,从土耳其哈塔伊省不同地方收集的发酵产品中鉴定出19种不同的乳酸菌。在分子鉴定中,通用引物对p806R/p8FPL和PEU7/DG74用于PCR扩增。之后,使用顺磁珠技术纯化的PCR产物通过桑格测序法进行测序。占分离株23.8%的优势菌种被鉴定为植物乳杆菌。作为乳酸菌的一项技术特性,在琼脂培养基中测试了42株乳酸菌分离株的胞外多糖产生能力,之后选择11株为阳性。当两株乳酸菌(弯曲乳杆菌SHA2-3B和棒状类肠球菌SHA6-3B)在添加了南瓜废料和乳清的MRS肉汤中培养时,它们具有较高的EPS产生能力。在添加10%南瓜废料的MRS肉汤中培养的棒状类肠球菌SHA6-3B中测定出最高的EPS含量(1750 mg/L葡萄糖当量)。此外,所产生的EPS样品通过傅里叶变换红外光谱(FTIR)和扫描电子显微镜(SEM)方法进行了验证。

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