Reconstructing ancient microbial fermentation genomes from the wine residues of Herod, Roman king of Judea

Abstract

The fortress of the Herodium, built towards the end of the first century BCE/ante Cristo, on the orders of Herod the Great, Roman client king of Judea, attests the expansion of Roman influence in the eastern Mediterranean. During archaeological excavations of the Herodium in 2017^[1], a winery was discovered on the ground floor of the palace, with an assortment of clay vessels in situ, including large dolia – clay fermentation vessels each capable of fermenting up to 300-400 L of wine. Thanks to the recent progresses in the field of paleogenomics^[2], we could analyse the organic material consistent with grape pomace at the bottom of these vessels, by extracting and sequencing the DNA using shotgun metagenomics and targeted capture, aiming for enrichment of DNA from fermentation associated microbes. After controlling for environmental and modern contamination, we successfully reconstructed and analysed the high-quality metagenome-assembled genomes (MAGs) from a phylogenetic and functional perspective. The phylogenetic signal of these ancient fermentation bacteria confirmed both their ancient origin and their affiliation to bacteria associated with wine fermentation. Gene functional analysis of the reconstructed fermentation-associated MAGs revealed an assortment of genes typically expected in lactic acid bacteria involved in wine fermentation, as well as genes involved in the production of wine spoilage compounds, especially in species typically viewed today as less desirable bacteria. Overall, our analysis brings a new appreciation of winemaking in Roman Judea and enriches our understanding of Roman accounts of flavouring wine with different herbs and aromatics, which may have been performed in part to mask the off-flavour compounds produced by bacterial wine spoilage genes.

Acknowledgements:

Werner Siemens Foundation (PALEOBIOTECHNOLOGY, funding M.B. and C.W.) and Deutsche Forschungsgemeinschaft (Balance of the Microverse, EXC 2051 #390713860, funding M.B. and C.W.)

References:

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