Molecular Cloning, Heterologous Expression, and Characterization of Ornithine Decarboxylase from Oenococcus oeni

Ornithine decarboxylase (ODC) is responsible for the production of putrescine, the major biogenic amine found in wine. Oenococcus oeni is the most important lactic acid bacterium in the winemaking process and is involved in malolactic fermentation. We report here the characterization of ODC from an O. oeni strain isolated from wine. Screening of 263 strains isolated from wine and cider from all over the world revealed that the presence of the odc gene appears to be strain specific in O. oeni. After cloning, heterologous expression in Escherichia coli, and characterization, the enzyme was found to have a molecular mass of 85 kDa and a pI of 6.2 and revealed maximal activity at pH 5.5 and an optimum temperature of 35°C. Kinetic studies showed that O. oeni ODC is specific for l-ornithine with a Km value of 1 mM and a Vmax of 0.57 U·mg−1. The hypothesis that cadaverine, which results from lysine decarboxylation, may be linked to putrescine production is not valid since O. oeni ODC cannot decarboxylate L-lysine. As no lysine decarboxylase was detected in any of the O. oeni genomes sequenced, cadaverine synthesis may result from another metabolic pathway. This work is the first characterization of an ODC from a lactic acid bacterium isolated from a fermented product.

Biogenic Amine Production by Oenococcus oeni Isolates from Malolactic Fermentation of Tempranillo Wine

In this article, we examine the production of biogenic amines, histamine, putrescine, tyramine, and cadaverine by 90 strains of Oenococcus oeni isolated from different cellars of Castilla-La Mancha (Spain) during wine malolactic fermentation. Amino biogenic capacity of strains was qualitatively analyzed on agar. After that, production of amines on a synthetic medium and on wine, and presence in strains of histidine, ornithine, and tyrosine decarboxylase genes were determined. Only two strains were able to produce histamine or putrescine, both on synthetic medium and wine. The presence of the corresponding genes in these strains was also confirmed. These results suggest that O. oeni does not significantly contribute to the overall biogenic amine content of wines. The main contribution of this work is the isolation of a putrescine-producing O. oeni strain that harbors the ornithine gene, since this gene appears to be rarely present in the genome of O. oeni.