Maria Gullo

Succession of Selected Strains of Acetobacter pasteurianus and Other Acetic Acid Bacteria in Traditional Balsamic Vinegar

ABSTRACT The application of a selected Acetobacter pasteurianus strain for traditional balsamic vinegar production was assessed. Genomic DNA was extracted from biofilms after enrichment cultures on GYC medium (10% glucose, 1.0% yeast extract, 2.0% calcium carbonate) and used for PCR/denaturing gradient gel electrophoresis, 16S rRNA gene sequencing, and enterobacterial repetitive intergenic consensus/PCR sequencing. Results suggested that double-culture fermentation is suitable for traditional balsamic vinegar acetification.

Occurrence and dominance of yeast species in sourdough.

Aims:  The aim of this work is to identify the dominant yeast species in homemade sourdoughs.

Traditional Balsamic Vinegar

The term ‘balsamic vinegar’ is frequently applied to describe sauces, condiments and dressings with particular sweet taste. In Italy there are two types of balsamic vinegar: ‘balsamic vinegar of Modena’ and ‘traditional balsamic vinegar’. The first is a flavoured wine vinegar obtained by blending cooked must and wine vinegar and, in some cases, by adding a small amount of caramel. Traditional balsamic vinegar (TBV) is made in Modena and Reggio Emilia with cooked grape must, through a three-step process: conversion of sugars to ethanol by yeasts; oxidation of ethanol to acetic acid by acetic acid bacteria (AAB); and, finally, at least 12 years of ageing. The final product is a highly dense, dark-brown aged vinegar, having a sweet and sour taste, fruity and complex in flavour.

Candida humilis--dominant species in sourdoughs for the production of durum wheat bran flour bread.

Yeasts present in the sourdough that is generally used for the production of durum wheat bran flour bread wereisolated and identified. Samples were taken during the rebuilding phase and at different intervals of time in order to monitor the population dynamics. The results obtained from the phenotypic studies were further confirmed by the molecular studies and enabled us to affirm that most of the strains, more than 95%, belong to the species Candida humilis. The dominance of C. humilis was steady in time. The isolations were carried out at sufficiently long intervals so that it was possible to ascertain that the conditions in which the sourdough is kept are fundamental to the microbiological stability of the dough.

Acetic Acid Bacteria: Physiology and Carbon Sources Oxidation

Acetic acid bacteria (AAB) are obligately aerobic bacteria within the family Acetobacteraceae, widespread in sugary, acidic and alcoholic niches. They are known for their ability to partially oxidise a variety of carbohydrates and to release the corresponding metabolites (aldehydes, ketones and organic acids) into the media. Since a long time they are used to perform specific oxidation reactions through processes called “oxidative fermentations”, especially in vinegar production. In the last decades physiology of AAB have been widely studied because of their role in food production, where they act as beneficial or spoiling organisms, and in biotechnological industry, where their oxidation machinery is exploited to produce a number of compounds such as l-ascorbic acid, dihydroxyacetone, gluconic acid and cellulose. The present review aims to provide an overview of AAB physiology focusing carbon sources oxidation and main products of their metabolism.

Diversity of Acetobacter pasteurianus strains isolated from solid-state fermentation of cereal vinegars.

Vinegar production is based on the acetification process by indigenous acetic acid bacteria (AAB). Among vinegar technologies, solid-state fermentation (SSF) processes are widespread in Asian countries to produce vinegar at small-scale. In this study, 21 AAB strains isolated from Chinese cereal vinegars produced by SSF collected in different regions of China were characterized by enterobacterial repetitive intergenic consensus (ERIC)–PCR fingerprinting. Isolates exhibited high degree of phenotypic variability as well as suitable traits for their uses as selected strains in SSF vinegar production (growth modality by superficial biofilm, no production of cellulose, ability to growth on ethanol media). 16S rRNA gene sequencing analysis of representative strains showed that strains of Acetobacter pasteurianus have a close association to cereal vinegars, whereas Gluconacetobacter europaeus population is not favoured. Selection of single or multiple strains culture within A. pasteurianus species was predicted in view of their application in SSF technology. This seems to be the first report showing phenotypic and genetic variability of AAB strains involved in SSF processes. Results can be exploited for the implementation of large-scale SSF processes by selected strains for vinegar production and other innovative biotechnological applications.

Evaluation and optimisation of bacterial genomic DNA extraction for no-culture techniques applied to vinegars

Direct genomic DNA extraction from vinegars was set up and suitability for PCR assays performed by PCR/DGGE and sequencing of 16S rRNA gene. The method was tested on 12 intermediary products of special vinegars, fruit vinegars and condiments produced from different raw materials and procedures. DNAs extraction was performed on pellets by chemical, enzymatic, resin mediated methods and their modifications. Suitable yield and DNA purity were obtained by modification of a method based on the use of PVP/CTAB to remove polyphenolic components and esopolysaccharides. By sequencing of bands from DGGE gel, Gluconacetobacter europaeus, Acetobacter malorum/cerevisiae and Acetobacter orleanensis were detected as main species in samples having more than 4% of acetic acid content. From samples having no acetic acid content, sequences retrieved from excised bands revealed high similarity with prokaryotes with no function on vinegar fermentation: Burkholderia spp., Cupriavidus spp., Lactococcus lactis and Leuconostoc mesenteroides. The method was suitable to be applied for no-culture study of vinegars containing polyphenols and esopolysaccharides allowing a more complete assessment of vinegar bacteria.

Acetic Acid Bacteria Taxonomy from Early Descriptions to Molecular Techniques

The exploitation of acetic acid bacteria (AAB) has a long history in fermentation processes and now represents an emerging field in biotechnological applications, especially with regard to the biosynthesis of useful chemicals with a potentially high economic value and, in food science, through the standardization of microbiological processes for the manufacture of both vinegar and other fermented beverages.

Biotechnological production of cellulose by acetic acid bacteria: current state and perspectives

Bacterial cellulose is an attractive biopolymer for a number of applications including food, biomedical, cosmetics, and engineering fields. In addition to renewability and biodegradability, its unique structure and properties such as chemical purity, nanoscale fibrous 3D network, high water-holding capacity, high degree of polymerization, high crystallinity index, light transparency, biocompatibility, and mechanical features offer several advantages when it is used as native polymer or in composite materials. Structure and properties play a functional role in both the biofilm life cycle and biotechnological applications. Among all the cellulose-producing bacteria, acetic acid bacteria of the Komagataeibacter xylinus species play the most important role because they are considered the highest producers. Bacterial cellulose from acetic acid bacteria is widely investigated as native and modified biopolymer in functionalized materials, as well as in terms of differences arising from the static or submerged production system. In this paper, the huge amount of knowledge on basic and applied aspects of bacterial cellulose is reviewed to the aim to provide a comprehensive viewpoint on the intriguing interplay between the biological machinery of synthesis, the native structure, and the factors determining its nanostructure and applications. Since in acetic acid bacteria biofilm and cellulose production are two main phenotypes with industrial impact, new insights into biofilm production are provided.

Oxidative fermentations and exopolysaccharides production by acetic acid bacteria: a mini review

Acetic acid bacteria are versatile organisms converting a number of carbon sources into biomolecules of industrial interest. Such properties, together with the need to limit chemical syntheses in favor of more sustainable biological processes, make acetic acid bacteria appropriate organisms for food, chemical, medical, pharmaceutical and engineering applications. At current, well-established bioprocesses by acetic acid bacteria are those derived from the oxidative pathways that lead to organic acids, ketones and sugar derivates. Whereas emerging applications include biopolymers, such as bacterial cellulose and fructans, which are getting an increasing interest for the biotechnological industry. However, considering the industrial demand of high performing bioprocesses, the production yield of metabolites obtained by acetic acid bacteria, is still not satisfying. In this paper we review the major acetic acid bacteria industrial applications, considering the current status of bioprocesses. We will also describe new biotechnological advances in order to optimize the industrial production, offering also an overview on future directions.