Department of Biology, Faculty of Mathematics and Natural Science, IPB University (Bogor Agricultural University), Gedung Biologi. Jalan Agatis Kampus IPB Dramaga, 16680, Bogor, Indonesia.
Faculty of Biotechnology, Atma Jaya Catholic University of Indonesia, Jalan Jenderal Sudirman 51, 12930, Jakarta, Indonesia.
BMC Res Notes. 2020 Dec 11;13(1):562. doi: 10.1186/s13104-020-05406-6.
Amplicon sequencing targeting 16S ribosomal RNA (rRNA) has been widely used to profile the microbial community from fermented food samples. However, polymerase chain reaction (PCR) steps on amplicon sequencing analysis and intragenomic heterogeneity within 16S rRNA are believed to contribute to bias in estimating microbial community composition. As potential paraprobiotics sources, a comprehensive profiling study of tempeh microbial ecology could contribute to tempeh product development. This study employed a shotgun metagenomic approach, where metagenome fragments from tempeh samples were sequenced directly for taxonomic and functional profiling analysis.
Taxonomic profiling showed that Proteobacteria, Firmicutes, and Bacteroidetes were the dominant phyla from the shotgun metagenomic analysis in all tempeh samples. In terms of composition, this shotgun metagenomic study revealed that Proteobacteria was the most abundant phylum. Functional profiling showed that iron complex outer-membrane recepter protein (KEGG ID: K02014) was the most transcribed gene based on this metagenomic analysis. The metagenome-assembled genomes (MAGs) results from the binning pipeline could reveal almost complete whole genome sequence of Lactobacillus fermentum, Enterococcus cecorum, Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii.
针对 16S 核糖体 RNA(rRNA)的扩增子测序已广泛用于从发酵食品样品中分析微生物群落。然而,扩增子测序分析中的聚合酶链反应(PCR)步骤和 16S rRNA 内的种内异质性被认为会导致对微生物群落组成的估计产生偏差。作为潜在的拟益生菌来源,对豆豉微生物生态学进行全面的分析研究有助于豆豉产品的开发。本研究采用了一种 shotgun 宏基因组学方法,其中直接对豆豉样品的宏基因组片段进行测序,用于分类和功能分析。
分类分析表明,在所有豆豉样品的 shotgun 宏基因组分析中,变形菌门、厚壁菌门和拟杆菌门是主要的门。在组成方面,这项 shotgun 宏基因组研究表明,变形菌门是最丰富的门。功能分析表明,铁复合物外膜受体蛋白(KEGG ID:K02014)是根据这项宏基因组分析转录最活跃的基因。基于 binning 管道的宏基因组组装基因组(MAGs)结果可以揭示完整的乳酸杆菌、肠球菌、大肠杆菌、肺炎克雷伯菌和鲍曼不动杆菌的全基因组序列。