First report on metagenomics and their predictive functional analysis of fermented bamboo shoot food of Tripura, North East India.

Moiya pansung, mileye amileye, moiya koshak, and midukeye are naturally fermented bamboo shoot foods of Tripura. The present study aimed to reveal the whole microbial community structure of naturally fermented moiya pangsung, mileye amileye, moiya koshak, and midukeye along with the prediction of microbial functional profiles by shotgun metagenomic sequence analysis. The metataxonomic profile of moiya pangsung, mileye amileye, moiya koshak, and midukeye samples showed different domains, viz., bacteria (97.70%) followed by the virus (0.76%), unclassified (0.09%), eukaryotes (1.46%) and archaea (0.05%). Overall, 49 phyla, 409 families, 841 genera, and 1,799 species were found in all the fermented bamboo shoot samples collected from different places of Tripura. Firmicutes was the most abundant phylum (89.28%) followed by Proteobacteria (5.13%), Bacteroidetes (4.38%), Actinobacteria (1.02%), and Fusobacteria (0.17%). Lactiplantibacillus plantarum was the most abundant species in moiya pangsung, mileye amileye, moiya koshak, and midukeye followed by Lactococcus lactis, Levilactobacillus brevis, Leuconostoc mesenteroides, Weissella paramesenteroides, Leuconostoc kimchii, Pediococcus pentosaceus, Leuconostoc gasicomitatum, and Lacticaseibacillus casei. A few phyla of fungus were found, viz., Ascomycota, Basidiomycota, and Glomeromycota, where Ascomycota was present in high abundance. Functional analysis of moiya pangsung, mileye amileye, moiya koshak, and midukeye metagenome revealed the genes for the synthesis and metabolism of a wide range of bioactive compounds including, various essential amino acids, and conjugated amino acids. The abundance profile and predictive analysis of fermented bamboo shoots revealed a huge plethora of essential microorganisms and KEGG analysis revealed genes for amino acid metabolism, pectin degradation, lipid metabolism, and many other essential pathways that can be essential for the improvement of nutritional and sensory qualities of the fermented bamboo shoot products.

Draft Genome Sequence of Lactiplantibacillus plantarum DMR09, Isolated from Spontaneously Fermented Dahi.

The draft genome sequence of Lactiplantibacillus plantarum DMR09, which was isolated from spontaneously fermented homemade cow dahi, was analyzed. The genome profile of L. plantarum DMR09 revealed 3,315,630 bp and a GC content of 44.5%, with 3,036 protein-encoding genes, 72 rRNA genes, and 65 tRNA genes.

Potential of Enterococcus faecium LM5.2 for lipopeptide biosurfactant production and its effect on the growth of maize (Zea mays L.).

This study characterized the lipopeptide biosurfactants produced by lactic acid bacteria isolated from milk and milk products, in addition to their effect on maize plant growth. The oil displacement test was performed as a primary screening method to select the BS producing bacteria. The strain LM5.2 had the maximum emulsification index of 45.1 ± 3, reduced the surface tension to 32.98 ± 0.23% among all the isolates, and efficiently produced 945.26 ± 4.62 mg/l of biosurfactants within 48 h in MRS broth under the optimum conditions. The isolate LM5.2 was identified using physiochemical tests and 16S rRNA gene sequencing as E. faecium. The biosurfactant was purified by TLC and identified as lipopeptide-like iturins and surfactins based on Rf values. Mass spectroscopy, NMR, and FTIR analysis also confirmed the biosurfactant's identity as the derivatives of iturin and surfactin. In vitro biosurfactant application significantly enhanced seedling growth at an optimal concentration of 450 µg/ml. Plant treatment (pot experiment) with E. faecium LM5.2 significantly promoted Zea mays growth compared to the control. To the best of our knowledge, this is the first report of lipopeptide biosurfactant production by E. faecium. Moreover, the study also showed that the biosurfactant and biosurfactant-producing E. faecium LM5.2 could be an eco-friendly plant growth-promoting agent.

Draft genome sequence of Lactobacillus plantarum strain DMR17 isolated from homemade cow dahi of Sikkim Himalayan region: an evaluation of lactate fermentation and secondary metabolism.

Lactobacillus plantarum DMR17 was isolated from homemade cow dahi of Sikkim Himalayan region of India. Here, we report the draft genome sequence of this strain. A total of 21,176,638 paired-end reads were obtained which were assembled into 65 contigs. The reference genome used was L. plantarum WCFS1. The genome size of DMR17 was 3,228,341 bp with G + C content of 46.25%. 3302 coding sequences were predicted including 68 tRNA and 67 rRNA genes. More than 88% of the total pre-processed reads from the sample were mapped to the reference genome. The identified coding proteins were classified into 27 functional categories based on COG classification. The genome was found to possess genes for lactate and mixed acid fermentation. The genome also showed the presence of genes for catabolism of aromatic compounds, phosphorous, and other metabolism. The genome information of L. plantarum DMR17 provides the basis for understanding the functional properties and to consider its use as a potential component of functional foods especially dahi.

Influence of biosurfactant producing Bacillus tequilensis LK5.4 isolate of kinema, a fermented soybean, on seed germination and growth of maize (Zea mays L.).

In this study, the lipopeptide biosurfactant was extracted, purified and characterized from the Bacillus isolate LK5.4 obtained from kinema samples of Sikkim. Plant growth-promoting property of the biosurfactant producing bacterium was also evaluated. Out of fifty-seven isolates, only ten were biosurfactant producer as determined by the oil displacement test. Bacillus isolate LK5.4 showed the maximum emulsification index (52.3 ± 0.02), reduced surface tension up to 40% and produced 754 mgL-1 biosurfactant in the nutrient broth. Based on 16S  rRNA gene sequencing, the isolate LK5.4 was identified as B. tequilensis. Biosurfactant was purified by Thin Layer Chromatography (TLC). Evaluation of the chemical characteristics by TLC, Liquid Chromatography-Mass Spectrometry, Fourier Transform Infrared Spectroscopy and Nuclear Magnetic Resonance Spectroscopy identified the biosurfactant as surfactin. The effect of different concentration of biosurfactant in maize seed germination was evaluated under in vitro condition. It showed the fastest growth of seedlings at 300 µg/ml biosurfactant solution. Similar results were shown by the potted plant experiment, where the soil was directly treated with biosurfactant producing bacterium LK5.4. The LK5.4 treated plants showed a mean height of 29.17 ± 0.47 cm and mean leaf length of 18.42 ± 0.17 cm while the mean height and mean length of the leaf were 15.48 ± 0.98 cm and 11.12 ± 0.40 cm respectively in the control plants. The treated plants had higher moisture content (68.48 ± 2.79%) than the control plants (50.53 ± 1.63%), which is because of higher bioadsorption in the treated plants. These results provided indirect evidence of plant growth-promoting property of the biosurfactant.

In situ fermentation dynamics during production of gundruk and khalpi, ethnic fermented vegetable products of the Himalayas.

Gundruk is a fermented leafy vegetable and khalpi is a fermented cucumber product, prepared and consumed in the Himalayas. In situ fermentation dynamics during production of gundruk and khalpi was studied. Significant increase in population of lactic acid bacteria (LAB) was found during first few days of gundruk and khlapi fermentation, respectively. Gundruk fermentation was initiated by Lactobacillus brevis, Pediococcus pentosaceus and finally dominated by Lb. plantarum. Similarly in khalpi fermentation, heterofermentative LAB such as Leuconostoc fallax, Lb. brevis and P. pentosaceus initiated the fermentation and finally completed by Lb. plantarum. Attempts were made to produce gundruk and khalpi using mixed starter culture of LAB previously isolated from respective products. Both the products prepared under lab condition had scored higher sensory-rankings comparable to market products.

Functional properties of lactic acid bacteria isolated from ethnic fermented vegetables of the Himalayas.

A total of 94 strains of Lactic acid bacteria (LAB), previously isolated from ethnic fermented vegetables and tender bamboo shoots of the Himalayas, were screened for functional properties such as acidification capacity, enzymatic activities, degradation of antinutritive factors and oligosaccharides, production of biogenic amines, hydrophobicity and adherence to mucus secreting HT29 MTX cells. Strong acidification and coagulation activities of LAB strains were recorded. Most of the LAB strains showed antimicrobial activities against the used indicator strains; however, only Lb. plantarum IB2 (BFE 948) isolated from inziangsang, a fermented leafy vegetable product, produced a bacteriocin against Staphylococcus aureus S1. LAB strains showed enzymatic activities and also degraded oligosaccharides. Almost all the strains of LAB were non-producers of biogenic amines except few strains. Some strains of Lb. plantarum showed more than 70% hydrophobicity. Adherence to the mucus secreting HT29 MTX cells was also shown by seven strains indicating their probiotic nature.

Phenotypic and genotypic identification of lactic acid bacteria isolated from ethnic fermented bamboo tender shoots of North East India.

Mesu, soidon, soibum and soijim are ethnic fermented bamboo tender shoot products prepared by the people in North East India. Microbiological analysis of mesu, soidon, soibum and soijim showed the population dominated by lactic acid bacteria (LAB) ranging up to 10(8) cfu g(-1). The phenotypic characterisation of predominant LAB isolated from the fermented bamboo shoot products was based on general morphology, physiological tests, API and Biolog systems. The genotypic characterisation of LAB was based on RAPD-PCR, rep PCR, species-specific PCR techniques, 16S rRNA gene sequencing and DNA-DNA hybridisation. Predominant functional LAB strains associated with the fermented bamboo shoot products were identified as Lactobacillus brevis, Lb. plantarum, Lb. curvatus, Pediococcus pentosaceus, Leuconostoc mesenteroides subsp. mesenteroides, Leuc. fallax, Leuc. lactis, Leuc. citreum and Enterococcus durans.

Identification of predominant lactic acid bacteria isolated from traditionally fermented vegetable products of the Eastern Himalayas.

Gundruk, sinki and khalpi are lactic-fermented vegetable products of Sikkim in India, and inziangsang is a fermented leafy vegetable product of Nagaland and Manipur in India. A total of 65 samples of gundruk (25), sinki (12), khalpi (25) and inziangsang (3) were analysed for microbial counts. The population of lactic acid bacteria (LAB) as well as aerobic mesophilic counts were at the level of 10(7) cfu g(-1). Yeasts were detected only in few samples of sinki and khalpi. No moulds were detected. In order to identify the predominating organisms, a total of 269 strains of LAB were isolated from gundruk, sinki, khalpi and inziangsang samples. The phenotypic characteristics of these strains were determined followed by genotyping using RAPD-PCR, repetitive element PCR and species-specific PCR techniques. The major representatives of the LAB involved in these fermentations were identified as Lactobacillus brevis, Lactobacillus plantarum, Pediococcus pentosaceus, Pediococcus acidilactici and Leuconostoc fallax.