Department of Molecular and Cellular Interactions, Flanders Institute of Biotechnology, Vrije Universiteit Brussel, Brussels, Belgium.
Appl Microbiol Biotechnol. 2010 Aug;87(6):2281-92. doi: 10.1007/s00253-010-2698-9. Epub 2010 Jun 18.
Cocoa bean fermentation is a spontaneous process involving a succession of microbial activities, starting with yeasts, followed by lactic acid bacteria and acetic acid bacteria. So far, all microbiological studies about cocoa bean fermentation were based on culture-dependent (isolation, cultivation, and identification), or, more recently, culture-independent (PCR-DGGE, or polymerase chain reaction denaturing gradient gel electrophoresis) methods. Using a metagenomic approach, total DNA was extracted from heap and box fermentations at different time points and from different locations (Ghana and Brazil, respectively) to generate a 16 S rDNA clone library that was sequenced. The sequencing data revealed a low bacterial diversity in the fermentation samples and were in accordance with the results obtained through culture-dependent and a second, culture-independent analysis (PCR-DGGE), suggesting that almost all bacteria involved in the fermentation process are cultivable. One exception was the identification by 16 S rDNA library sequencing of Gluconacetobacter species of acetic acid bacteria that were not detected by the two other approaches. The presence of Enterobacteriaceae related to Erwinia/Pantoea/Tatumella, as revealed by 16 S rDNA library sequencing, suggests an impact of these bacteria on fermentation.
可可豆发酵是一个自发的过程,涉及一系列微生物活动,首先是酵母,然后是乳酸菌和醋酸菌。到目前为止,所有关于可可豆发酵的微生物学研究都是基于培养依赖型(分离、培养和鉴定),或者最近,基于培养非依赖型(PCR-DGGE,或聚合酶链反应变性梯度凝胶电泳)方法。使用宏基因组学方法,从堆肥和箱式发酵中提取不同时间点和不同地点(分别来自加纳和巴西)的总 DNA,以生成 16S rDNA 克隆文库并进行测序。测序数据显示发酵样品中的细菌多样性较低,与培养依赖型和第二种培养非依赖型分析(PCR-DGGE)的结果一致,表明参与发酵过程的几乎所有细菌都是可培养的。一个例外是通过 16S rDNA 文库测序鉴定出的醋酸菌属中的 Gluconacetobacter 种,这两种方法都没有检测到。通过 16S rDNA 文库测序揭示的与 Erwinia/Pantoea/Tatumella 相关的肠杆菌科的存在表明这些细菌对发酵有影响。
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