A novel culture medium for Oenococcus oeni malolactic starter production
Malolactic fermentation is essential in wine quality. One of the strategies used to control this fermentation involves the inoculation of selected lactic acid bacteria, mainly Oenococcus oeni. Laboratory media usually produce large amounts of biomass, but with little or no adaptability to wine. We propose a culture medium to grow and pre-adapt O. oeni cells, and the steps to scale-up production. To achieve this objective, 27 different media were tested. All contained grape must and wine, and nutritional supplements in order to benefit bacterial growth. Those media contained different ethanol levels, pH values, and grape must concentrations. The optimized culture medium named Oenococcus Prod…
Microbial resources and sparkling wine differentiation : state of the arts
Consumers’ increasing interest in sparkling wine has enhanced the global market’s demand. The pro-technological yeasts strains selected for the formulation of microbial starter cultures are a fundamental parameter for exalting the quality and safety of the final product. Nowadays, the management of the employed microbial resource is highly requested by stakeholders, because of the increasing economic importance of this oenological sector. Here, we report an overview of the production processes of sparkling wine and the main characterisation criteria to select Saccharomyces and non-Saccharomyces strains appropriate for the preparation of commercial starter cultures dedicated to the primary a…
Immobilisation of yeasts on oak chips or cellulose powder for use in bottle-fermented sparkling wine
[EN] Sparkling wine production comprises two successive fermentations performed by Sacharomyces cerevisiae strains. This research aimed to: develop yeast immobilisation processes on two wine-compatible supports; study the effects of yeast type (IOC 18-2007 and 55A) and the immobilisation support type (oak chips and cellulose powder) on the fermentation kinetics, the deposition rate of lees and the volatile composition of the finished sparkling wine; compare the fermentation parameters of the wines inoculated with immobilised or non-immobilised cells. Proper immobilisation of yeast on oak chips and cellulose powder was demonstrated by electron microscopy. Total sugar consumption occurred in …
A Metagenomic-Based Approach for the Characterization of Bacterial Diversity Associated with Spontaneous Malolactic Fermentations in Wine
This study reports the first application of a next generation sequencing (NGS) analysis. The analysis was designed to monitor the effect of the management of microbial resources associated with alcoholic fermentation on spontaneous malolactic consortium. Together with the analysis of 16S rRNA genes from the metagenome, we monitored the principal parameters linked to MLF (e.g., malic and lactic acid concentration, pH). We encompass seven dissimilar concrete practices to manage microorganisms associated with alcoholic fermentation: Un-inoculated must (UM), pied-de-cuve (PdC), Saccharomyces cerevisiae (SC), S. cerevisiae and Torulaspora delbrueckii co-inoculated and sequentially inoculated, as…
Requirement of the Lactobacillus casei MaeKR two-component system for L-malic acid utilization via a malic enzyme pathway.
ABSTRACTLactobacillus caseican metabolizel-malic acid via malolactic enzyme (malolactic fermentation [MLF]) or malic enzyme (ME). Whereas utilization ofl-malic acid via MLF does not support growth, the ME pathway enablesL. caseito grow onl-malic acid. In this work, we have identified in the genomes ofL. caseistrains BL23 and ATCC 334 a cluster consisting of two diverging operons,maePEandmaeKR, encoding a putative malate transporter (maeP), an ME (maeE), and a two-component (TC) system belonging to the citrate family (maeKandmaeR). Homologous clusters were identified inEnterococcus faecalis,Streptococcus agalactiae,Streptococcus pyogenes, andStreptococcus uberis. Our results show that ME is …
The use of core-shell high-performance liquid chromatography column technology to improve biogenic amine quantification in wine
BACKGROUND HPLC column technology has been improved, providing better resolution of closely eluting compounds, better analyte sensitivity, and shorter analysis times. The core-shell technology columns offer a faster analysis through the use of shorter columns without compromising resolution. The aim of this work was to improve the methods for determination of biogenic amines (BAs) in wine using the new HPLC PFP core-shell column technology. RESULTS Two different elution programs were designed to quantify BAs with the core-shell PFP column. Program I flow rate was 2 mL min−1. The total elution time was 10 min. In elution program II, the flow rate was 0.8 mL min−1 and the total elution time w…
A new fear in wine: Isolation of Staphylococcus epidermidis histamine producer
Abstract Biogenic amines (BAs) are undesirable compounds in all foods and beverages because they may induce food-borne intoxications when consumed at high concentrations. The aim of this study was to identify the organism responsible for the synthesis of histamine in a Tempranillo red wine from Ribera de Duero D.O. (Denomination of origin), Spain. Bacteria present in wines after malolactic fermentation were isolated, identified and typed. Four strains of the species Oenococcus oeni and two strains of Staphylococcus epidermidis were found. None of the O. oeni strains produced histamine, cadaverine or putrescine, but one of the S. epidermidis strains (Pa) was able to produce all three in synt…
Starter cultures as biocontrol strategy to prevent Brettanomyces bruxellensis proliferation in wine
Brettanomyces bruxellensis is a common and significant wine spoilage microorganism. B. bruxellensis strains generally detain the molecular basis to produce compounds that are detrimental for the organoleptic quality of the wine, including some classes of volatile phenols that derive from the sequential bioconversion of specific hydroxycinnamic acids such as ferulate and p-coumarate. Although B. bruxellensis can be detected at any stage of the winemaking process, it is typically isolated at the end of the alcoholic fermentation (AF), before the staring of the spontaneous malolactic fermentation (MLF) or during barrel aging. For this reason, the endemic diffusion of B. bruxellensis leads to c…
Use of autochthonous yeasts and bacteria in order to control Brettanomyces bruxellensis in wine
Biocontrol strategies for the limitation of undesired microbial developments in foods and beverages represent a keystone toward the goal of more sustainable food systems. Brettanomyces bruxellensis is a wine spoilage microorganism that produces several compounds that are detrimental for the organoleptic quality of the wine, including some classes of volatile phenols. To control the proliferation of this yeast, sulfur dioxide is commonly employed, but the efficiency of this compound depends on the B. bruxellensis strain; and it is subject to wine composition and may induce the entrance in a viable, but nonculturable state of yeasts. Moreover, it can also elicit allergic reactions in humans. …
Isolation and characterisation of autochthonous Saccharomyces cerevisiae from 'Pago' Merlot wines of Utiel-Requena (Spain) origin
[EN] Background and Aims To achieve both the control of the undesirable yeasts and the improvement of the characteristics of Merlot 'Pago' wines, Saccharomyces cerevisiae yeast diversity was investigated in a spontaneous 'Pago' Merlot fermentation from the Utiel-Requena region (Spain). Isolates of S. cerevisiae were characterised by a holistic procedure using the same Merlot grape must from which they were isolated. Methods and Results Yeasts were identified by internal transcribed spacer analysis and typed by HinfI mDNA restriction analysis. Growth and metabolic characteristics of the isolates were determined by laboratory-scale (50 L) fermentations of Merlot must and the composition of th…
Selection of indigenous yeast strains for the production of sparkling wines from native Apulian grape varieties.
We report the first polyphasic characterization of native Saccharomyces cerevisiae in order to select candidate strains for the design of starter cultures tailored for Apulian sparkling wines obtained from local grape variety. In addition, it is the first survey in our region that propose the selection of autochthonous starter cultures for sparkling wine i) including a preliminary tailored genotypic and technological screening, and ii) monitoring analytical contribution during secondary fermentation in terms of volatile compounds (VOCs). Furthermore, we exploit the potential contribute of autochthonous cultures throughout the productive chain, including the possible improvement of base wine…
Lowering histamine formation in a red Ribera del Duero wine (Spain) by using an indigenous O. oeni strain as a malolactic starter
This study demonstrates for the first time that a non-commercial selected autochthonous O. oeni strain has been used to conduct malolactic fermentation (MLF) while lowering histamine formation in the same winery. Lactic acid bacteria (LAB) were isolated from 13 vats before and after spontaneous MLF at the Pago de Carraovejas winery from the Ribera del Duero region (Spain). Only O. oeni were present, typed and characterized, and both histamine producer and non-producers existed. From the non-producers, one strain was selected to become a starter according to its genetic profile, prevalence in the different wines in the winery, resistance to alcoholic degree, resistance to high polyphenolic c…
Technological properties of Lactobacillus plantarum strains isolated from grape must fermentation
Malolactic fermentation (MLF) is a secondary fermentation in wine that usually takes place during or at the end of alcoholic fermentation. Lactobacillus plantarum is able to conduct MLF (particularly under high pH conditions and in co-inoculation with yeasts) and some strains are commercially used as MLF starter cultures. Recent evidences suggest a further use of selected L. plantarum strains for the pre-alcoholic acidification of grape must. In this study, we have carried out an integrated (molecular, technological, and biotechnological) characterization of L. plantarum strains isolated from Apulian wines in order to combine the two protechnological features (MLF performances and must acid…
Saccharomyces cerevisiae and Oenococcus oeni immobilized in different layers of a cellulose/starch gel composite for simultaneous alcoholic and malolactic wine fermentations
Abstract The production of a two-layer composite biocatalyst for immobilization of two different microorganisms for simultaneous alcoholic and malolactic fermentation (MLF) of wine in the same bioreactor is reported. The biocatalyst consisted of a tubular delignified cellulosic material (DCM) with entrapped Oenococcus oeni cells, covered with starch gel containing the alcohol resistant and cryotolerant strain Saccharomyces cerevisiae AXAZ-1. The biocatalyst was found effective for simultaneous low temperature alcoholic fermentation resulting to conversion of malic acid to lactic acid in 5 days at 10 °C. Improvement of wine quality compared with wine fermented with S. cerevisiae AXAZ-1 immob…