Metagenome analysis to fingerprint bacterial and fungi communities in tempe

Abstract

Tempe is an Indonesian traditional fermented food produced using Rhizopus as a starter culture. In practice, however, the starter culture as well as fermentation processes would involved a polymicrobial fermentation, which generated a unique tempe flavor and texture. This condition makes Indonesian tempe as one of the most complex fermented food, while at the same time would make it difficult to scale up tempe production with uniform quality and consistency. Because of complex microbial communities involved, cultureindependent methods or metagenomic analysis would be an essential tool to unravel this complex microbial fermentation. The aim of this study was to compare a number of tempe microbial communities based on Amplified Ribosomal Intergenic Sequence Analysis (ARISA). Fresh tempe samples were obtained from tempe producers in Java and Moluccas. 16S rRNA gene libraries and DNA sequencing were employed to analyze further the nature of microbial diversity in two selected tempe samples. The results of our study showed that different tempe producer possessed different Bacterial ARISA (BARISA) or fungi ARISA (FARISA) profiles. However, BARISA profiles were found to be more discriminative than FARISA, and therefore BARISA would be more useful for tempe genetic fingerprint or barcoding. Bacterial communities in two tempe samples (EMP and SDJD) were further investigated employing DNA sequence analysis of their 16S rRNA gene libraries. Sequences closely related to Klebsiella sp were found to be the majority among bacterial communities present in SDJD sample, while sequences closely related to Acetobacter sp. were found as predominant isolates in EMP samples. These significant differences in bacterial communities might reflect differences in tempe flavor or texture associated with their unique production or locality.