Three novel structural phenomena in the cellular ontogeny of Oenococcus oeni from northern China.

Stress resistance and growth are important aspects to consider when engineering Oenococcus oeni strains for winemaking. We identified 3 previously unreported structural phenomena in the cell ontogeny of O. oeni sampled in northern China. We show that budding and binary fission (BBF) occur simultaneously in the growth process; that a novel 'pomegranate-shaped structure' (PSS) occurs mainly in the stationary and death phases; and that symbiosis and cyclical phenomena (SCP) occur throughout the various cell growth phases. These observations add to the current knowledge of the cell growth process of O. oeni. BBF, PSS, and SCP sufficiently describe the characteristics of the cellular ontogeny of O. oeni. We highlight a newly identified structure that explains the complex cell growth process. These findings will help understand the growth and development of O. oeni, supplementing the knowledge base of the established phases and providing new perspectives into its complex growth patterns.

Exopolysaccharide (EPS) synthesis by Oenococcus oeni: from genes to phenotypes.

Oenococcus oeni is the bacterial species which drives malolactic fermentation in wine. The analysis of 50 genomic sequences of O. oeni (14 already available and 36 newly sequenced ones) provided an inventory of the genes potentially involved in exopolysaccharide (EPS) biosynthesis. The loci identified are: two gene clusters named eps1 and eps2, three isolated glycoside-hydrolase genes named dsrO, dsrV and levO, and three isolated glycosyltransferase genes named gtf, it3, it4. The isolated genes were present or absent depending on the strain and the eps gene clusters composition diverged from one strain to another. The soluble and capsular EPS production capacity of several strains was examined after growth in different culture media and the EPS structure was determined. Genotype to phenotype correlations showed that several EPS biosynthetic pathways were active and complementary in O. oeni. Can be distinguished: (i) a Wzy-dependent synthetic pathway, allowing the production of heteropolysaccharides made of glucose, galactose and rhamnose, mainly in a capsular form, (ii) a glucan synthase pathway (Gtf), involved in ß-glucan synthesis in a free and a cell-associated form, giving a ropy phenotype to growth media and (iii) homopolysaccharide synthesis from sucrose (α-glucan or ß-fructan) by glycoside-hydrolases of the GH70 and GH68 families. The eps gene distribution on the phylogenetic tree was examined. Fifty out of 50 studied genomes possessed several genes dedicated to EPS metabolism. This suggests that these polymers are important for the adaptation of O. oeni to its specific ecological niche, wine and possibly contribute to the technological performance of malolactic starters.

Genetic and physiological studies on Oenococcus oeni PsuI in response to ethanol stress.

Malolactate fermentation (MLF), which is known to decreases total acidity and improves the stability and quality of cider is conducted by Oenococcus oeni; the principal microorganism responsible for MLF under stress conditions. Understanding O. oeni physiology in stress conditions can be used to generate tools based on molecular and physiological approaches allowing more precise characterization of strains. Regarding intracellular protein, the results showed an increase in the levels of amino acids under ethanol stress. To study the expressed genes under ethanol stress, one gene were sequenced. An outer-membrane lipoprotein carrier protein precursor, Lo1A was expressed under ethanol stress conditions. Scanning electron microscopy was used to study the effect of ethanol stress on cell morphology. SEM revealed aggregation of bacterial cells as the level of ethanol increases in culture medium in comparison to controls.