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利用变性梯度凝胶电泳分析韩国泡菜及其原料的微生物群落结构

Microbial Community Structure of Korean Cabbage Kimchi and Ingredients with Denaturing Gradient Gel Electrophoresis.

作者信息

Hong Sung Wook, Choi Yun-Jeong, Lee Hae-Won, Yang Ji-Hee, Lee Mi-Ai

机构信息

World Institute of Kimchi, Gwangju 61755, Republic of Korea.

出版信息

J Microbiol Biotechnol. 2016 Jun 28;26(6):1057-62. doi: 10.4014/jmb.1512.12035.

Abstract

Kimchi is a traditional Korean fermented vegetable food, the production of which involves brining of Korean cabbage, blending with various other ingredients (red pepper powder, garlic, ginger, salt-pickled seafood, etc.), and fermentation. Recently, kimchi has also become popular in the Western world because of its unique taste and beneficial properties such as antioxidant and antimutagenic activities, which are derived from the various raw materials and secondary metabolites of the fermentative microorganisms used during production. Despite these useful activities, analysis of the microbial community present in kimchi has received relatively little attention. The objective of this study was to evaluate the bacterial community structure from the raw materials, additives, and final kimchi product using the culture-independent method. Specifically, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the 16S rRNA partial sequences of the microflora. One primer set for bacteria, 341F(GC)-518R, reliably produced amplicons from kimchi and its raw materials, and these bands were clearly separated on a 35-65% denaturing gradient gel. Overall, 117 16S rRNA fragments were identified by PCR-DGGE analysis. Pediococcus pentosaceus, Leuconostoc citreum, Leuconostoc gelidum, and Leuconostoc mesenteroides were the dominant bacteria in kimchi. The other strains identified were Tetragenococcus, Pseudomonas, Weissella, and uncultured bacterium. Comprehensive analysis of these microorganisms could provide a more detailed understanding of the biologically active components of kimchi and help improve its quality. PCR-DGGE analysis can be successfully applied to a fermented food to detect unculturable or other species.

摘要

泡菜是一种传统的韩国发酵蔬菜食品,其制作过程包括用盐水腌制大白菜,与各种其他配料(红辣椒粉、大蒜、生姜、盐渍海鲜等)混合,然后进行发酵。最近,泡菜因其独特的口味以及抗氧化和抗诱变活性等有益特性而在西方世界也受到欢迎,这些特性源自制作过程中使用的各种原材料和发酵微生物的次生代谢产物。尽管有这些有益活性,但对泡菜中存在的微生物群落的分析相对较少受到关注。本研究的目的是使用非培养方法评估原材料、添加剂和最终泡菜产品中的细菌群落结构。具体而言,聚合酶链反应 - 变性梯度凝胶电泳(PCR - DGGE)用于分析微生物群落的16S rRNA部分序列。一套用于细菌的引物341F(GC)-518R能够可靠地从泡菜及其原材料中产生扩增子,并且这些条带在35 - 65%的变性梯度凝胶上清晰分离。总体而言,通过PCR - DGGE分析鉴定出117个16S rRNA片段。戊糖片球菌、柠檬明串珠菌、嗜冷明串珠菌和肠膜明串珠菌是泡菜中的优势细菌。鉴定出的其他菌株有嗜盐四联球菌、假单胞菌、魏斯氏菌和未培养细菌。对这些微生物的综合分析可以更详细地了解泡菜的生物活性成分,并有助于提高其品质。PCR - DGGE分析可以成功应用于发酵食品以检测不可培养或其他物种。

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