AUTHOR=Gopal Murali , Shil Sandip , Gupta Alka , Hebbar K. B. , Arivalagan M. 

TITLE=Metagenomic Investigation Uncovers Presence of Probiotic-Type Microbiome in Kalparasa® (Fresh Unfermented Coconut Inflorescence Sap)

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.662783

DOI=10.3389/fmicb.2021.662783

ISSN=1664-302X

ABSTRACT=<p>The phloem sap tapped from unopened inflorescence (spadix) of coconut palm using a novel collecting device, “coco-sap chiller,” has been branded Kalparasa<sup>®</sup> (henceforth as <italic>Kalparasa</italic> in the text) to distinguish its properties not found in sap harvested by traditional methods. To know its hitherto unidentified microbiome profile, we employed high-throughput sequencing to uncover the bacteriome and mycobiome in fresh and 12-h fermented samples. Fresh <italic>Kalparasa</italic> had a pH of 7.2, which dropped to 4.5 after 12 h, signifying fermentation of the sap. Diversity analysis indicated fresh <italic>Kalparasa</italic> having higher bacterial species than the fermented one. Contrary to this, fresh sap had lower fungal/yeast diversity than the fermented sample. Fresh <italic>Kalparasa</italic> had relatively higher abundance of probiotic-type <italic>Leuconostoc</italic> genus followed by equal proportions of <italic>Gluconobacter</italic>, <italic>Acetobacter</italic>, and <italic>Fructobacillus</italic>. The 12-h fermented <italic>Kalparasa</italic> showed a significant increase in <italic>Gluconobacter</italic> with a sharp decrease in <italic>Leuconostoc</italic>. Mycobiome data revealed fresh <italic>Kalparasa</italic> to be preponderant in <italic>Saccharomyces</italic> and <italic>Hanseniaspora</italic> genera of yeasts while the fermented sap had higher representation of <italic>Hanseniaspora</italic> and <italic>Cortinarius</italic> and lesser <italic>Saccharomyces</italic>. This suggested that the fermentation of <italic>Kalparasa</italic> was probably driven by symbiotic culture of bacteria and yeasts (SCOBY), particularly acetic acid bacteria and non-<italic>Saccharomyces</italic> yeasts. The bacteriome-function predictions highlighted the enrichment of glycerophospholipid, ABC transporters, purine, and pyrimidine metabolisms. Based on our findings, <italic>Kalparasa</italic> containing large population of <italic>Leuconostoc mesenteroides</italic>, <italic>Fructobacillus fructosus</italic>, <italic>Saccharomyces cerevisiae</italic>, and <italic>Hanseniaspora guilliermondii</italic> can be promoted as a healthy “unfermented” plant edible food containing live probiotic-type microbiome during its consumption.</p>