Optimization of bacterial cellulose production by Komagataeibacter sucrofermentans in synthetic media and agrifood side streams supplemented with organic acids and vitamins.

The effect of thiamine (TA), ascorbic acid (AA), citric acid, and gallic acid (GA) on bacterial cellulose (BC) production by Komagataeibacter sucrofermentans, in synthetic (Hestrin and Schramm, HS) and natural substrates (industrial raisins finishing side stream extract, FSSE; orange juice, OJ; green tea extract, GTE), was investigated. The Response Surface Methodology was found reliable for BC yield prediction and optimization. Higher yields were achieved in the FSSE substrates, especially those supplemented with AA, TA, and GA (up to 19.4 g BC/L). The yield in the non-fortified substrates was 1.1-5.4 and 11.6-15.7 g/L, in HS and FSSE, respectively. The best yield in the natural non-fortified substrate FSSE-OJ-GTE (50-20-30 %), was 5.9 g/L. The porosity, crystallinity, and antioxidant properties of the produced BC films were affected by both the substrate and the drying method (freeze- or oven-drying). The natural substrates and the process wastewaters can be further exploited towards added value and sustainability. Take Home Message Sentence: Raisin and citrus side-streams can be efficiently combined for bacterial cellulose production, enhanced by other vitamin- and phenolic-rich substrates such as green tea.

Production and In Situ Modification of Bacterial Cellulose Gels in Raisin Side-Stream Extracts Using Nanostructures Carrying Thyme Oil: Their Physicochemical/Textural Characterization and Use as Antimicrobial Cheese Packaging.

We report the production of BC gels by Komagataeibacter sucrofermentans in synthetic (Hestrin and Schramm; HS) and natural media (raisin finishing side-stream extracts; RFSE), and their in situ modification by natural zeolite (Zt) and activated carbon (AC) nanostructures (NSs) carrying thyme oil (Th). The NS content for optimum BC yield was 0.64 g/L for both Zt-Th (2.56 and 1.47 g BC/L in HS and RFSE, respectively), and AC-Th (1.78 and 0.96 g BC/L in HS and RFSE, respectively). FTIR spectra confirmed the presence of NS and Th in the modified BCs, which, compared to the control, had reduced specific surface area (from 5.7 to 0.2-0.8 m2/g), average pore diameter (from 264 to 165-203 Å), cumulative pore volume (from 0.084 to 0.003-0.01 cm3/g), crystallinity index (CI) (from 72 to 60-70%), and crystallite size (from 78 to 72-76%). These values (except CI and CS), slightly increased after the use of the BC films as antimicrobial coatings on white cheese for 2 months at 4 °C. Tensile properties analysis showed that the addition of NSs resulted in a decrease of elasticity, tensile strength, and elongation at break values. The best results regarding an antimicrobial effect as cheese coating were obtained in the case of the RFSE/AC-Th BC.

Sweet Wine Production from the Side-Stream of Industrial Corinthian Currant Processing: Product Quality, Antioxidant Capacity, and Volatilome.

In the frame of efforts to add value to the Mediterranean currant cultivation and processing sectors, which is essential for their sustainability, sweet wine production is proposed from the finishing side-stream (FSS) of premium quality Corinthian currants, involving complete fermentation using an alcohol-tolerant yeast followed by (i) the addition of FSS to extract sugars or (ii) syrup made from FSS to adjust sweetness. Wine was also made by (iii) ceasing fermentation at the desired sugar level by ethanol addition. The non-fortified wines had 15.2-15.5% ethanol, 115-145 g/L residual sugar, 7.2-7.6 g/L titratable acidity, low volatile acidity (VA; <0.33 g/L), 280-330 mg/L phenolic content (TPC) (as gallic acid), and 23.8-35.6 mg/L antioxidant capacity (AC) (as ascorbic acid). In total, 160 volatiles were identified by SPME GC-MS, including compounds derived from the grapes, the raisin drying, and the fermentation process. The non-fortified wines had better characteristics (mainly VA, AC, and TPC) than the fortified wine, while sweetness adjustment by FSS is the simplest and lowest cost method since it does not involve ethanol or syrup addition. The proposed methods can lead to good quality sweet wines with a characteristic fruity (grape/raisin) flavor that could be commercialized as specialty raisin beverages or liqueurs.

Nutritional Improvements of Sourdough Breads Made with Freeze-Dried Functional Adjuncts Based on Probiotic Lactiplantibacillus plantarum subsp. plantarum and Pomegranate Juice.

New types of sourdough breads are proposed, made with freeze-dried sourdough adjuncts based on: (i) Lactiplantibacillus plantarum subsp. plantarum ATCC 14917, a potential probiotic (LP) alone or (ii) with the addition of unfermented pomegranate juice (LPPO) and (iii) pomegranate juice fermented by the same strain (POLP). Physicochemical, microbiological, and nutritional characteristics (in vitro antioxidant capacity, AC, total phenolics, TPC, and phytate content) of the breads were evaluated and compared with commercial sourdough bread. All adjuncts performed well; the best results being those obtained by POLP. Specifically, the highest acidity (9.95 mL of 0.1 M NaOH) and organic acid content (3.02 and 0.95 g/kg, lactic and acetic acid, respectively) as well as better resistance to mold and rope spoilage (12 and 13 days, respectively) were observed for POLP3 bread (sourdough with 6% POLP). Significant nutritional improvements were observed by all adjuncts, in terms of TPC, AC, and phytate reduction (103 mg gallic acid/100 g, 232 mg Trolox/100 g, and 90.2%, respectively, for POLP3). In all cases, the higher the amount of adjunct, the better the results. Finally, the good sensory properties of the products indicate the suitability of the proposed adjuncts for sourdough breadmaking, while their application in freeze-dried, powdered form can facilitate commercial application.

Assessment of the Physicochemical, Antioxidant, Microbial, and Sensory Attributes of Yogurt-Style Products Enriched with Probiotic-Fermented Aronia melanocarpa Berry Juice.

The aim() of this study was to create() various formulations of yogurt enriched with freeze()-dried adjuncts, namely() (i) probiotic Lactobacillus plantarum ATCC 14917 culture(), and (ii) L. plantarum ATCC 14917 fermented black chokeberry juice, along with a commercial() starter culture(). The goal was to enhance() functionality and optimize the nutritional() value() of the products. These new yogurt-style() formulations were subsequently() compared with commercially produced yogurt. All products demonstrated() favorable() physicochemical properties, and the probiotic strain() consistently() maintained viable() levels exceeding 7 log() cfu/g throughout() the entire() storage() period(). The fermented milk produced with the adjunct-free L. plantarum cells, as well as the yogurt produced with the proposed() lactobacilli-fermented chokeberry juice, exhibited the highest lactic acid() production() (1.44 g/100 g yogurt by the end of storage()). Levels of syneresis were observed at lower() values() in yogurt produced with freeze()-dried fermented chokeberry juice. Yogurts prepared() with the lactobacilli-fermented freeze()-dried chokeberry juice displayed elevated total() phenolic content() and antioxidant capacity() (25.74 µg GAE/g and 69.05 µmol TE/100 g, respectively()). Furthermore, sensory tests revealed a distinctive() fruity flavor() in samples incorporating fermented juice. The results demonstrate() that probiotic L. plantarum-fermented chokeberry juice enhances() both the antioxidant capacity() and the viability of beneficial() bacteria() in yogurt while it can be readily() applied and commercialized, especially in the form of a freeze()-dried formulation.

Vinegar Production from Corinthian Currants Finishing Side-Stream: Development and Comparison of Methods Based on Immobilized Acetic Acid Bacteria.

Fruit wastes and side-streams can be used for vinegar production to create added value for the agri-food sector and enhance farmer incomes and local economies. In this study, methods for vinegar production by wild and selected acetic acid bacteria (the quick starter Acetobacter aceti and the acid-resistant Komagataeibacter europaeus), free (FC) and immobilized (IC) on a natural cellulosic carrier, are proposed using sweet wine made from the industrial finishing side-stream (FSS) of Corinthian currants as raw material. The results showed all cultures can produce vinegar with 46.65 ± 5.43 g/L acidity, from sweet FSS wine containing 5.08 ± 1.19% alcohol. The effect of immobilization was more obvious in the case of the selected culture, presenting better acetification efficiency, both fresh and after cold storage for 2 months. The vinegars had an antioxidant capacity of 263.5 ± 8.4 and 277.1 ± 6.7 mg/L (as ascorbic acid) and phenolic content 333.1 ± 12.0 and 222.2 ± 2.9 mg/L (as gallic acid) (for FC and IC, respectively). They also had a rich volatilome (140 compounds identified by SPME GC-MS), with higher percentages of esters identified in vinegars made by IC. The results are encouraging for vinegar production with IC of a mixed A. aceti and K. europaeus culture.

Health Promoting Properties of Cereal Vinegars.

Vinegar has been used for its health promoting properties since antiquity. Nowadays, these properties are investigated, scientifically documented, and highlighted. The health benefits of vinegar have been associated with the presence of a variety of bioactive components such as acetic acid and other organic acids, phenolic compounds, amino acids, carotenoids, phytosterols, vitamins, minerals, and alkaloids, etc. These components are known to induce responses in the human body, such as antioxidant, antidiabetic, antimicrobial, antitumor, antiobesity, antihypertensive, and anti-inflammatory effects. The diversity and levels of bioactive components in vinegars depend on the raw material and the production method used. Cereal vinegars, which are more common in the Asia-Pacific region, are usually made from rice, although other cereals, such as millet, sorghum, barley, malt, wheat, corn, rye, oats, bran and chaff, are also used. A variety of bioactive components, such as organic acids, polyphenols, amino acids, vitamins, minerals, alkaloids, melanoidins, butenolides, and specific compounds such as γ-oryzanol, tetramethylpyrazine, γ-aminobutyric acid, etc., have been associated with the health properties of cereal vinegars. In this work, the bioactive components and the related health effects of cereal vinegars are reviewed, and the most recent scientific literature is presented and discussed.

Corinthian currants finishing side-stream: Chemical characterization, volatilome, and valorisation through wine and baker's yeast production-technoeconomic evaluation.

The industrial currants finishing generates a considerable amount of side-stream (FSS) with great potential for biotechnological exploitation. The chemical composition of FSS generated from the premium quality Vostitsa currants was studied. Its use for wine making (at low temperatures, using both free and immobilized yeast) combined with baker's yeast production (with minor nutrient supplementation), is also proposed. Analysis showed that FSS has a rich volatilome (including Maillard reaction/lipid degradation products), increased antioxidant capacity, and total lipid and phenolic contents, compared to the marketable product (currants). However, acidity levels and the presence of specific volatiles (such as acetate esters and higher alcohols) may be indicative of microbial spoilage. The wines made from FSS were methanol free and contained higher levels of terpenes (indicating hydrolysis of bound forms) and fermentation-derived volatiles, compared to FSS. A preliminary technoeconomic analysis for integrated wine/baker's yeast industrial production, showed that the investment is realistic and worthwhile.

Evaluation of Pediococcus pentosaceus SP2 as Starter Culture on Sourdough Bread Making.

In the present study, a novel Pediococcus pentosaceus SP2 strain, recently isolated from kefir grains, was evaluated as a starter culture in sourdough bread making. The novel starter was applied in fresh, freeze-dried, and freeze-dried immobilized (on wheat bran) form. The type of culture (fresh, freeze-dried, immobilized cells) influenced the bread characteristics. Specifically, the application of freeze-dried immobilized cells led to higher total titratable acidity (TTA) values (9.81 mL NaOH N/10), and the produced bread presented higher resistance to mold and rope spoilage. Moreover, the produced sourdough breads were significantly better in terms of pH, TTA, organic acids content, and resistance to mold and rope spoilage, compared to breads made with a commercial, wild microbiota, sourdough. The organic acids content was also significantly higher than the commercial sourdough sample (2.93 g/kg lactic acid; 1.01 g/kg acetic acid). Determination of volatile compounds through solid-phase microextraction (SPME) gas chromatography/mass spectrometry (GC/MS) analysis and sensorial assessments indicated no significant differences between the tested sourdough breads.

Functional pomegranate beverage production by fermentation with a novel synbiotic L. paracasei biocatalyst.

The recently isolated Lactobacillus paracasei K5 (a potential probiotic) was immobilized on delignified wheat bran (DWB) and was used to produce a functional pomegranate beverage. Fermentations were carried out for 24 h at different pH values (3.3, 3.6, and 3.9), and the fermented juices were stored for 4 weeks at 4 °C. In all cases, the immobilized biocatalyst was shown to be efficient for pomegranate juice fermentation, as indicated by analysis of sugars and organic acids, and ethanol production was maintained at low levels (0.5-1% v/v). The sugars consumed during fermentation were used mainly for cell mass production, while DWB exerted a prebiotic effect, protecting and enhancing the viability of L. paracasei. Optimum viability (9.8-11.7 log cfu/mL) was achieved at pH 3.9. The phenolic content and headspace volatiles were increased after fermentation and storage (112 ±â€¯17-213 ±â€¯30 mg GAE/100 mL and 27-46 compounds, respectively), indicating potential to produce good quality, synbiotic pomegranate beverages.

Bacterial Cellulose Production Using the Corinthian Currant Finishing Side-Stream and Cheese Whey: Process Optimization and Textural Characterization.

The aim of this work was to develop bioprocesses to produce a high-value microbial product, bacterial cellulose (BC), utilizing the industrial side-stream of Corinthian currants finishing (CFS), with/without the addition of N-sources and cheese whey, and at various process conditions (temperature, pH level, and sugar concentration). For the optimization of BC production, the response surface methodology based on the central composite design was applied. Among the possible retrieved combinations, the most ideal conditions for BC in CFS extracts supplemented with N-source were 28 °C, pH 6.42, and 46.24 g/L concentration of sugars. In a similar manner, the best conditions for BC production in CFS/whey mixtures were pH 6.36, 50.4% whey percentage in the mixture, and 1.7% yeast extract. The textural characteristics of the produced BC, at different times of production and using different drying methods, were studied by scanning electron microscopy, X-ray diffractometry, porosimetry, Fourier-transform infrared spectroscopy, and thermogravimetric/differential thermal analysis, revealing increased porosity of BC compared with delignified cellulosic materials of plant origin, and a level of crystallinity that depended on the BC production time. The proposed methodology can be used to produce foods with potential prebiotic properties, using the highly nutritious CFS and the abundant cheese whey effluent as raw materials.

Effect of a novel Lactobacillus paracasei starter on sourdough bread quality.

The novel Lactobacillus paracasei K5 strain, recently isolated from Greek cheese, was evaluated as potential sourdough bread starter. Breads were made using different amounts of L. paracasei sourdoughs as well as traditional sourdough for comparison. Quality characteristics of the breads (acidity and rising) were examined, as well as rope spoilage through macroscopic observations and molecular analysis (PCR-DGGE). The highest acidity levels (3.15 g lactic acid and 1.13 g acetic acid per kg of bread) and better resistance to rope spoilage were observed when bread contained 30% w/w L. paracasei K5 sourdough. Spoilage in the L. paracasei K5 breads was observed at 15-16 days, 5 days later than the control breads. In addition, L. paracasei K5 sourdough improved the bread sensory properties, as reflected by consumer preference and GC/MS analysis of aroma volatiles. Therefore, L. paracasei K5 can be successfully used for sourdough bread making with good quality and extended shelf-life.

Application of A Novel Potential Probiotic Lactobacillus paracasei Strain Isolated from Kefir Grains in the Production of Feta-Type Cheese.

In the present study 38 lactic acid bacteria strains were isolated from kefir grains and were monitored regarding probiotic properties in a series of established in vitro tests, including resistance to low pH, resistance to pepsin and pancreatin, and tolerance to bile salts, as well as susceptibility against common antibiotics. Among them, the strain SP3 displayed potential probiotic properties. Multiplex PCR analysis indicated that the novel strain belongs to the paracasei species. Likewise, the novel strain (Lactobacillus paracasei SP3) was applied as a starter culture for Feta-type cheese production. Feta-type cheese production resulted in significantly higher acidity; lower pH; reduced counts of coliforms, yeasts and fungi; and improved quality characteristics compared with cheese samples produced with no starter culture. Finally, it is highlighted that the application of the novel strain led to Feta-type cheese production with improved overall quality and sensory characteristics.

Potential of the Probiotic Lactobacillus Plantarum ATCC 14917 Strain to Produce Functional Fermented Pomegranate Juice.

In this research survey the application of probiotic strain Lactobacillus plantarum ATCC 14917 in pomegranate juice fermentation is sought. Pomegranate juice was fermented for 24 h and then it was stored 4 for 4 weeks. Cell viability retained in high levels after the 24 h of fermentation and storage for 4 weeks (above 8.8 log cfu/mL), while fermented pomegranate juice was scored better at the 4th week of storage compared to non-fermented pomegranate juice. The probiotic strain was effective regarding lactic acid fermentation as was proved through sugar and organic acids analysis. Concentration of ethanol was maintained at low levels (0.3⁻1% v/v). Fermented pomegranate juice contained more and in higher percentages desirable volatile compounds (alcohols, ketones and esters) even at the 4th week of cold storage compared to non-fermented juice. Antioxidant activity (150.63 mg Trolox equivalent (TE)/100 mL at the 2nd week) and total phenolic content (206.46 mg gallic acid equivalents (GAE)/100 mL at the 2nd week) were recorded in higher levels for all the storage time compared to non-fermented juice.

Employment of L. paracasei K5 as a Novel Potentially Probiotic Freeze-Dried Starter for Feta-Type Cheese Production.

In the present study, a novel potentially probiotic Lactobacillus paracasei strain, previously isolated from dairy products, was evaluated as a starter culture of Feta-type cheese production. Targeting industrial applications, the starter culture was applied as a ready-to-use freeze-dried culture that was either free or immobilized. The immobilized biocatalyst composed of Lactobacillus paracasei K5 cells absorbed within delignified wheat bran prebiotic carrier. All produced cheeses were compared with cheese manufactured by renin enzyme. Several parameters that affect acceptability, quality and shelf-life of Feta-type cheese were investigated, including microbial populations, physicochemical characteristics and cheese volatiles through 90 days of ripening and storage. Survival of L. paracasei K5 remained in high levels (≥6.0 log cfu/g) after the 90th day of cheese production, as recorded by combining microbiological enumeration and strain-specific multiplex PCR analysis. The use of the freeze-dried novel starter culture (free or immobilized) enhanced the aromatic profile of Feta-type cheeses. Finally, the use of the potentially synbiotic immobilized biocatalyst further improved aromatic characteristics of produced cheese and decrease of possible spoilage or pathogenic microorganisms. These findings indicate the potential industrial use of freeze-dried L. paracasei K5 as starter culture for the production of good-quality functional Feta-type cheese.

Growth Capacity of a Novel Potential Probiotic Lactobacillus paracasei K5 Strain Incorporated in Industrial White Brined Cheese as an Adjunct Culture.

In this study, a novel potential probiotic strain Lactobacillus paracasei K5, previously isolated from traditional Greek Feta cheese and kefir grains, was evaluated as an adjunct culture for industrial white brined cheese production. Targeting industrial applications, apart from free cell cultures, a novel ready-to-use freeze-dried immobilized biocatalyst was prepared. The biocatalyst composed of L. paracasei K5 cells immobilized on delignified wheat bran prebiotic carrier and was freeze-dried without cryoprotectants. The adjunct free or immobilized culture was added separately without prior adaptation during white brined cheese manufacture and the produced cheeses were compared with commercial white brined cheeses. Several parameters that affect the acceptability and quality of the cheeses, including microbial populations, physicochemical parameters, volatile by-products and organoleptic characteristics, were analyzed through 70 days of storage. Results showed that the viability of the adjunct culture added either free or immobilized remained in high levels (7 to 8 log cfu/g) during maturation and storage. In addition, all white brined cheeses with the adjunct probiotic culture showed a sharp decrease in spoilage and pathogenic microorganisms such as enterobacteria, salmonella, staphylococci and coliforms during cheese maturation, especially when compared with the commercial white brined cheeses. Finally, after maturation time exceeded, all cheeses were characterized as safe for human consumption. Cheeses volatile compounds were significantly enhanced by the incorporated immobilized biocatalysts. These findings indicate the potential industrial use of freeze-dried ready-to-use immobilized lactobacilli as reinforcement adjunct cultures for the production of good-quality functional cheese products. PRACTICAL APPLICATION: The launch on market of novel foods developed by the incorporation of functional ingredients provides potential benefits to consumers' diet and new business opportunities for producers. Probiotic food products are one significant category of functional foods. Thus, this study focused on the development of a novel ready-to-use freeze-dried potential probiotic biocatalyst for functional white brined cheese production. The potential industrial application of such biocatalysts is highlighted by their incorporation as adjunct cultures that resulted in good-quality functional cheeses.

Technological Development of Brewing in Domestic Refrigerator Using Freeze-Dried Raw Materials.

Development of a novel directly marketable beer brewed at low temperature in a domestic refrigerator combined with yeast immobilization technology is presented in this study. Separately, freeze-dried wort and immobilized cells of the cryotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 on tubular cellulose were used in low-temperature fermentation (2, 5 and 7 °C). The positive effect of tubular cellulose during low-temperature brewing was examined, revealing that freeze-dried immobilized yeast cells on tubular cellulose significantly reduced the fermentation rates in contrast to freeze-dried free cells, although they are recommended for home-made beer production. Immobilization also enhanced the yeast resistance at low-temperature fermentation, reducing the minimum brewing temperature value from 5 to 2 °C. In the case of high-quality beer production, the effect of temperature and initial sugar concentration on the fermentation kinetics were assessed. Sensory enrichment of the produced beer was confirmed by the analysis of the final products, revealing a low diacetyl concentration, together with improved polyphenol content, aroma profile and clarity. The proposed process for beer production in a domestic refrigerator can easily be commercialized and applied by dissolving the content of two separate packages in tap water; one package containing dried wort and the other dried immobilized cells on tubular cellulose suspended in tap water.

Porous cellulose as promoter of oil production by the oleaginous yeast Lipomyces starkeyi using mixed agroindustrial wastes.

Enhanced single cell oil (SCO) production by the oleaginous yeast Lipomyces starkeyi DSM 70296, immobilised on delignified porous cellulose, is reported. Pure glucose media were initially used. The effects of substrate pH and treatment temperature were evaluated, showing that 30°C and pH 5.0 were the optimum conditions for SCO production by the immobilised yeast. The immobilisation technique led to increased lipid accumulation and cell growth by 44% and 8%, respectively, in the glucose media, compared to free cells in suspension. This positive effect was also shown when low concentration mixed agro-industrial waste suspensions were used as substrates, leading to 85% enhanced SCO production in comparison with free cells. Higher fatty acid (HFA) analysis showed that yeast immobilisation led to increased formation of unsaturated HFAs (6%) and reduced saturated HFAs (5%) compared to free cells.

Scale-up for esters production from straw whiskers for biofuel applications.

Delignified wheat straw was fermented by a mixed bacterial anaerobic culture obtained from a UASB reactor to produce organic acids (OAs). Kissiris was used as immobilization carrier in a 2-compartment 82L bioreactor filled with 17L of fermentation broth for the first 7 fermentation batches and up to 40L for the subsequent batches. The amount of straw used was 30g/L and the temperature was set at 37°C for all experiments. The total OAs reached concentrations up to 17.53g/L and the produced ethanol ranged from 0.3 to 1mL/L. The main OAs produced was acetic acid (6-8g/L) and butyric acid (3-8g/L). The OAs were recovered from the fermentation broth by a downstream process using 1-butanol, which was the solvent with the best recovery yields and also served as the esterification alcohol. The enzymatic esterification of OAs resulted to 90% yield.

Downstream extraction process development for recovery of organic acids from a fermentation broth.

The present study focused on organic acids (OAs) recovery from an acidogenic fermentation broth, which is the main problem regarding the use of OAs for production of ester-based new generation biofuels or other applications. Specifically, 10 solvents were evaluated for OAs recovery from aqueous media and fermentation broths. The effects of pH, solvent/OAs solution ratios and application of successive extractions were studied. The 1:1 solvent/OAs ratio showed the best recovery rates in most cases. Butyric and isobutyric acids showed the highest recovery rates (80-90%), while lactic, succinic, and acetic acids were poorly recovered (up to 45%). The OAs recovery was significantly improved by successive 10-min extractions. Alcohols presented the best extraction performance. The process using repeated extractions with 3-methyl-1-butanol led to the highest OAs recovery. However, 1-butanol can be considered as the most cost-effective option taking into account its price and availability.