Tempe is an Indonesian fermented food prepared by fermenting dehulled cooked soybeans with Rhizopus oligosporus.Many types of bacteria are also involved during tempe fermentation, and one of these is Klebsiella spp. Some isolates of K. pneumoniae produces vitamin B12 in tempe but it has also been classified as an opportunistic pathogen. For this reason Klebsiella spp. in tempe is important to be studied. The aim of this study was to investigate the genetic diversity of Klebsiella spp. from tempe employing ERIC-PCR method. Sixty-one isolates of Klebsiella have been isolated from sixteen tempe producers in Bogor, Jakarta, Malang, Tengerang, Bandung and Cianjur. 63F and 1387R primers were used to amplify 16S rDNA sequences, and 1R and 1F primers were used for ERIC analysis. The results of this research showed that sixty-one strains of Klebsiella were clustered into 17 groups. Based on ERIC-PCR analysis, isolates of Klebsiella could be grouped into different profiles which some of these groups consisted of isolates with identical ERIC-PCR profiles. Several identical ERIC-PCR profiles were found in tempe from the same producer. There was no correlation observed between genetic similarity among isolates with the origin of tempe.

Fermented cassava or “Tape” is one of traditional Indonesian fermented food. The quality of “Tape” is determined by microorganisms involved during fermentation process. It was reported that Bacillus subtilis determined the quality of cassava “Tape”. The most common way to identify species is by using 16S rRNA gene. This gene contains conserved regions as unique sequence which is relative among species. It has been widely used as a reliable molecular marker for phylogeny identification. Therefore, the aim of this research was to study diversity of amylase-producing Bacillus spp. from “Tape” based on 16S rRNA gene sequences. Bacillus spp. were isolated from “Tape” from several area in Indonesia i.e. Jakarta, Bandung, Cianjur, Subang, Rangkas Bitung, and Kediri. Amplification of 16S rRNA gene used 63f and 1387r primers. This research showed that based on 16S rRNA gene sequences, twenty-six of amylase-producing Bacillus spp. isolates were divided into four groups. All isolates were identified as species either B. megaterium, B. subtilis, B. amyloliquefaciens, or B. thuringiensis

Aims: Our previous study demonstrated that Klebsiella IIEMP-3 associated with tempeh was genetically different from those of medical isolates. In addition to the whole genome sequence of Klebsiella IIEMP-3, the draft genome sequence of another isolate, i.e. IWJB-6 was employed for comparison. In this study, the details of the virulence genes and unique gene in both Klebsiella isolates were compared employing in silico and in vitro analysis. Methodology and results: Whole genome of Klebsiella IIEMP-3 and IWJB-6 were annotated to investigate the virulence factor. Klebsiella IIEMP-3 and IWJB-6 were obtained from tempeh producers in Bogor, West Java-Indonesia. Genome sequences were analyzed employing BLAST Ring Image Generator (BRIG) software. The results showed that all of the samples, including isolates IIEMP-3 and IWJB-6 did not harbor rmpA, i.e. DNA sequence for K. pneumoniae virulence factor. Conclusion, significance and impact of study: Klebsiella could be found in almost all tempeh samples from Indonesia and could be harmless for human due to the absence of rmpA and other virulence-associated genes. The significance of this study showed that IIEMP-3 and IWJB-6 isolates were more closely related to K. variicola. However, K. variicola At-22 harbored sdsA gene which is lacking in those two tempeh isolates. Combined with PCR analysis for specific gene/s; our study suggested that isolates from Indonesian tempeh were closely related to K. variicola, and proposed to be designated as K. variicola subsp. tempehensis.

Klebsiella pneumoniae strain IIEMP-3, isolated from Indonesian tempeh, is a vitamin B12-producing strain that exhibited a different genetic profile from pathogenic isolates. Here we report the draft genome sequence of strain IIEMP-3, which may provide insights on the nature of fermentation, nutrition, and immunological function of Indonesian tempeh.

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