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In the present investigation, the potential of various newly isolated strains which belong to the Enterobacteriaceae family to produce 2,3-butanediol (BDO), an important bio-based compound, was studied. The most interesting strain, namely Klebsiella oxytoca FMCC-197, was selected for further investigation. Commercial (raw) sucrose or molasses, which are important agro-industrial surpluses, were employed as carbon sources for most of the trials performed. Different fermentation parameters (viz. incubation te4mperature, utilization of different carbon sources, substrate inhibition, aeration) were tested to optimize the process. Fermentations under non-aseptic conditions were also conducted to investigate the potential of growth of the strain K. oxytoca FMCC-197 to surpass the growth of other microorganisms in the culture medium and produce BDO. Besides BDO production, in trials in which molasses was employed as the sole carbon source, significant color removal was observed simultaneously with the production of microbial metabolites. The very high BDO concentration ≈115 g L−1 was reported in approximately 64 h during a fed-batch bioreactor experiment, using sucrose and molasses as carbon sources at 30 °C, reaching a conversion yield (YBDO) of 0.40 g g−1 and a productivity rate (PBDO) of 1.80 g L−1 h−1, while similar results were also obtained at 37 °C. The strain demonstrated remarkable results in non-previously sterilized media, as it produced 58.0 g L−1 in 62 h during a fed-batch bioreactor experiment, while the potential to decolorize molasses-based substrates over 40% was also recorded. From the results obtained it is shown that this wild-type strain can be used in large-scale microbial BDO production using various raw materials as fermentative substrates. The wastewater derived after BDO fermentation by K. oxytoca FMCC-197 can be disposed relatively safely into the environment. [ABSTRACT FROM AUTHOR]

Fish waste accounts for almost one-third of the total fish production annually. The main objective of this study was to upcycle fish by-products to produce biodegradable packaging materials. Fish protein concentrate (FPC) was extracted from gilthead seabream by-catch (flesh and skin). FPC (3%) and gelatin (3%) were used to produce film-forming solutions. The films were produced according to the solvent casting method. The produced films were tested as packaging materials via the determination of different film properties. The wettability of the packaging materials was characterized based on the determination of the contact angle. Water vapor permeability was evaluated using the ASTM E96/E96M standardized method. The evaluation of mechanical properties was based on the Young's modulus, tensile strength, and elongation at break. Color was measured using a CIELab system. The incorporation of FPC into the produced membranes resulted in a reduced contact angle from 108.5° to 90.6°; however, both films were characterized as hydrophobic materials. Films supplemented with FPC had lower tensile strength values compared to pure gelatin films, but higher elongation values without statistically significant differences. The color parameters (L,a,b) indicated that gelatin films and FPC–gelatin films were colorless and transparent (L > 90), an important parameter for food packaging materials. The production of biodegradable packaging materials from FPC and gelatin may effectively reduce petroleum-based plastics under the circular economy model. [ABSTRACT FROM AUTHOR]

In this study, the use of vinification by-products in broiler diets, as a sustainable and promising way of exploiting them, was examined. In particular, the potential use of ground grape pomace (GGP), wine lees extract (WYC) and grape stem extract (PE) in broiler diets was examined. Growth performance parameters, the weight of selected internal organs, meat quality traits, fatty acid profiles of breast meat and selected haematological parameters were determined. Two hundred and forty one-day-old broilers were assigned to four treatments with four replicate pens of fifteen broilers. There was one control treatment (CON), fed a basal diet, and the GGP, WYC and PE treatments, fed a basal diet supplemented with 25 g/kg GGP, 2 g/kg WYC and 1 g starch including 100 mg pure stem extract/kg PE, respectively. The duration of the experiment was 42 days. The average body weight gain during the starter, grower and finisher stages did not differ among treatments. Similarly, the feed intake, FCR and carcass yield did not show a significant difference. The weight of the internal organs was also similar among treatments. Some positive differences were observed in colour traits of meat and in haematological parameters. In the GGP group, saturated (SFAs) and unsaturated fatty acids (USFAs) were lower and higher, respectively, compared to the CON, WYC and PE groups. Vinification by-products seem to be a promising feed additive in broiler diets providing a sustainable approach to grape waste management without affecting broiler performance. [ABSTRACT FROM AUTHOR]

Lignocellulosic waste-streams are resources worth investigating to produce value-added bioproducts due to their low-cost, renewability, versatility, and abundance. This study evaluated five Zygomycota strains as potential oleaginous microorganisms for direct bioconversion of cakes derived from the vegetable-oil-milling industry into microbial oil. Solid-state fermentation (SSF) conditions in tray bioreactors revealed that temperature, moisture content, and substrate affected lipid production and fatty acids composition. Employing SSF, under optimal conditions (30 °C, 65% initial moisture content), and substrate supplementation with nitrogen sources increased lipid productivity 1.1-fold on palm kernel cake (PKC) substrate. Supplementation strategies using glycerol, molasses, or mineral solution did nοt favor lipid production. When cotton seed cake (CoSC) was applied, C. echinulata was able to produce 304.1 mg/gfs of biomass with a lipid content of 40.2% (w/w) in tray bioreactors. Scaling-up SSF using CoSC in packed bed reactors led to the highest biomass production (320.5 mg/gfs), while lipids production was unaffected. Then, microbial oil was enzymatically converted into polyol esters achieving the highest conversion yield of 80% after 2 h. Physicochemical properties of polyol esters demonstrated their potential utilization as biolubricants. The present study showed the perspective of bioconverting industrial side streams into microbial oil and a route for sustainable synthesis of oleochemicals. [ABSTRACT FROM AUTHOR]

Although carotenoids generally possess antimicrobial and antioxidant properties, the in vivo synergistic action of carotenoid blends derived from plant-based by-products has not been thoroughly studied. Therefore, the carotenoid characterization and antimicrobial potential of Citrus reticulata extract as well as the impact of this carotenoid-rich extract (CCE) dietary supplementation on the performance, meat quality, and immune-oxidative status of broiler chickens were determined. One hundred and twenty one-day-old hatched chicks (Ross 308) were allocated to two dietary groups, with four replicate pens of 15 birds each. Birds were fed either a basal diet (CON) or the basal diet supplemented with 0.1% CCE (25 mg carotenoid extract included in 1 g of soluble starch) for 42 d. β-Cryptoxanthin, β-Carotene, Zeaxanthin, and Lutein were the prevailing carotenoid compounds in the Citrus reticulata extract. The CCE feed additive exerted inhibitory properties against both Gram-positive (Staphylococcus aureus) and negative (Klebsiella oxytoca, Escherichia coli, and Salmonella typhimurium) bacteria. Both the broiler performance and meat quality did not substantially differ, while the breast muscle malondialdehyde (MDA) concentration tended to decrease (p = 0.070) in the CCE-fed broilers. The inclusion of CCE decreased the alanine aminotransferase and MDA concentration, and the activity of glutathione peroxidase, while the activity of superoxide dismutase was increased in the blood. Catalase and NADPH oxidase 2 relative transcript levels were significantly downregulated in the livers of the CCE-fed broilers. Additionally, Interleukin 1β and tumor necrosis factor (TNF) relative transcript levels were downregulated in the livers of the CCE- fed broilers, while TNF and interferon γ (IFNG) tended to decrease in the spleens and bursa of Fabricius, respectively. The present study provided new insights regarding the beneficial properties of carotenoids contained in Citrus reticulata in broilers’ immune-oxidative status. These promising outcomes could be the basis for further research under field conditions. [ABSTRACT FROM AUTHOR]

The utilization of crude glycerol, generated as a by-product from the biodiesel production process, for the production of high value-added products represents an opportunity to overcome the negative impact of low glycerol prices in the biodiesel industry. In this study, the biochemical behavior of Yarrowia lipolytica strains FMCC Y-74 and FMCC Y-75 was investigated using glycerol as a carbon source. Initially, the effect of pH value (3.0-7.0) was examined to produce polyols, intracellular lipids, and polysaccharides. At low pH values (initial pH 3.0-5.0), significant mannitol production was recorded. The highest mannitol production (19.64 g L-1) was obtained by Y. lipolytica FMCC Y-74 at pH = 3.0. At pH values ranging between 5.0 and 6.0, intracellular polysaccharides synthesis was favored, while polyols production was suppressed. Subsequently, the effect of crude glycerol and its concentration on polyols production was studied. Y. lipolytica FMCC Y-74 showed high tolerance to impurities of crude glycerol. Initial substrate concentrations influence polyols production and distribution with a metabolic shift toward erythritol production being observed when the initial glycerol concentration (Gly0) increased. The highest total polyols production (=56.64 g L-1) was obtained at Gly0 adjusted to -120 g L-1. The highest polyols conversion yield (0.59 g g-1) and productivity (4.36 g L-1 d-1) were reached at Gly0 = 80 g L-1. In fed-batch intermittent fermentation with glycerol concentration remaining ≤60 g L-1, the metabolism was shifted toward mannitol biosynthesis, which was the main polyol produced in significant quantities (=36.84 g L-1) with a corresponding conversion yield of 0.51 g g-1. [ABSTRACT FROM AUTHOR]

Oleogelation is an emerging technology to structure oils, which can be widely used to substitute saturated and trans fats. Extra virgin olive oil is widely recognized for its high nutritional value, but its utilization in oleogel production is currently limited. In this study, extra virgin olive oil was utilized for the production of a novel oleogel using wax esters derived from soybean fatty acid distillate (SFAD), a byproduct of industrial soybean oil refining. Different concentrations (7%, 10%, 20%, w/w) of SFAD-wax esters were used to evaluate the minimum concentration requirement to achieve oleogelation. Analyses of the mechanical properties of oleogel showed a firmness of 3.8 N, which was then reduced to around 2.1-2.5 N during a storage period of 30 days at 4 °C. Rheological analysis demonstrated that G' is higher than G' at 20-27 °C, which confirms the solid properties of the oleogel at this temperature range. Results showed that SFAD was successfully utilized for the oleogelation of olive oil, resulting in a novel oleogel with desirable properties for food applications. This study showed that industrial fatty side streams could be reused for the production of value-added oleogels with novel food applications. [ABSTRACT FROM AUTHOR]

The utilization of fermentation media derived from waste and by-product streams from biodiesel and confectionery industries could lead to highly efficient production of bacterial cellulose. Batch fermentations with the bacterial strain Komagataeibacter sucrofermentans DSM (Deutsche Sammlung von Mikroorganismen) 15973 were initially carried out in synthetic media using commercial sugars and crude glycerol. The highest bacterial cellulose concentration was achieved when crude glycerol (3.2 g/L) and commercial sucrose (4.9 g/L) were used. The combination of crude glycerol and sunflower meal hydrolysates as the sole fermentation media resulted in bacterial cellulose production of 13.3 g/L. Similar results (13 g/L) were obtained when flour-rich hydrolysates produced from confectionery industry waste streams were used. The properties of bacterial celluloses developed when different fermentation media were used showed water holding capacities of 102-138 g·water/g·dry bacterial cellulose, viscosities of 4.7-9.3 dL/g, degree of polymerization of 1889.1-2672.8, stress at break of 72.3-139.5 MPa and Young's modulus of 0.97-1.64 GPa. This study demonstrated that by-product streams from the biodiesel industry and waste streams from confectionery industries could be used as the sole sources of nutrients for the production of bacterial cellulose with similar properties as those produced with commercial sources of nutrients. [ABSTRACT FROM AUTHOR]

Biobased pigments are environmentally friendly alternatives to synthetic variants with an increased market demand. Production of pigments via fermentation is a promising process, yet optimization of the production yield and rate is crucial. Herein, we evaluated the potential of Penicillium purpurogenum to produce biobased pigments. Optimum sugar concentration was 30 g/L and optimum C:N ratio was 36:1 resulting in the production of 4.1–4.5 AU (namely Pigment Complex A). Supplementation with ammonium nitrate resulted in the production of 4.1–4.9 AU (namely Pigment Complex B). Pigments showed excellent pH stability. The major biopigments in Pigment Complex A were N-threonyl-rubropunctamin or the acid form of PP-R (red pigment), N-GABA-PP-V (violet pigment), PP-O (orange pigment) and monascorubrin. In Pigment Complex B, a novel biopigment annotated as N-GLA-PP-V was identified. Its basic structure contains a polyketide azaphilone with the same carboxyl-monascorubramine base structure as PP-V (violet pigment) and γ-carboxyglutamic acid (GLA). The pigments were not cytotoxic up to 250 μg/mL. [ABSTRACT FROM AUTHOR]

Simple Summary: The valorization of grape byproducts appears to be a sustainable bioeconomic strategy that could promote the substitution of synthetic with natural antioxidant compounds in the food and feed industry. The nutritional physiology of broiler chickens requires special attention to the interactions between nutrients and antioxidant mechanisms since the stressor signals of factory farming could impair the immune status, resulting in detrimental effects on broilers' performance. The objective of this study was to assess the inclusion of grape byproducts (grape pomace, wine lees, and stem extract) on the transcriptional profiling of genes regulating the immune system in the liver, bursa of Fabricius, and spleen. The dietary supplementation of grape byproducts rich in polyphenolic compounds decreased the mRNA levels of the predominant pro-inflammatory receptor in the liver, while in the spleen, the stem extract diet upregulated the aforementioned receptor. Upregulation of interleukin 8 was observed in the bursa of Fabricius and spleen of the stems extract-fed broilers. Although grape byproducts depicting a sustainable source of bioactive compounds with vast antioxidant potential, there were unveiled preliminary insights for immune stimulation at the transcriptional level. The valorization of vinification byproducts portrays a promising bioprocess for the enrichment of animals' diet with bioactive compounds, such as polyphenols, which could regulate the immune response. Therefore, the impact of dietary grounded grape pomace (GGP), wine lees extract (WYC), and grape stem extract (PE) on the relative transcript level of immune related genes of broiler chickens were examined. Two hundred forty, one-day-old as hatched (male/female) chicks (Ross 308) were allocated to four dietary groups, with four replicate pens each with 15 birds. Birds were fed either a basal diet (CON) or the basal diet supplemented with 2.5% GGP, or 0.2% WYC, or 0.1% PE for 42 d. The relative expression of immune-related genes was investigated using a real-time PCR platform. The mRNA levels of Toll-like Receptor 4 (TLR4) were downregulated (p = 0.039) in the liver of broilers fed the GGP-containing diet compared to the CON, while in the spleen of PE-fed broilers, TLR4 was significantly upregulated (p = 0.043). The mRNA levels of interleukin 8 (IL8) tended to upregulate (p = 0.099) in the bursa of Fabricius and were significantly increased (p = 0.036) in the spleen of broilers fed the PE diet. Vinification byproducts depict a promising sustainable source of polyphenols for the poultry feed industry, but more research is needed under field conditions. [ABSTRACT FROM AUTHOR]

Vinification by-products display great potential for utilization as feed additives rich in antioxidant compounds. Thus, the effect of dietary ground grape pomace (GGP), wine lees extract rich in yeast cell walls (WYC), and grape stem extracts (PE) on the relative expression of several genes involved in liver oxidative mechanisms and the oxidative status of the blood and breast muscle of broiler chickens was investigated. In total, 240 one-day-old as hatched chicks (Ross 308) were assigned to four treatments, with four replicate pens and 15 birds in each pen. Birds were fed either a basal diet (CON) or a basal diet supplemented with 25 g/kg GGP, or 2 g/kg WYC, or 1 g starch including 100 mg pure stem extract/kg (PE) for 42 days. The polyphenolic content of vinification by-products was determined using an LC-MS/MS library indicating as prevailing compounds procyanidin B1 and B2, gallic acid, caftaric acid, (+)-catechin, quercetin, and trans-resveratrol. Body weight and feed consumption were not significantly affected. The relative transcript level of GPX1 and SOD1 tended to increase in the liver of WYC-fed broilers, while NOX2 tended to decrease in the PE group. SOD activity in blood plasma was significantly increased in WYC and PE compared to the CON group. The total antioxidant capacity measured with FRAP assay showed significantly higher values in the breast muscle of PE-fed broilers, while the malondialdehyde concentration was significantly decreased in both WYC- and PE-fed broilers compared to the CON group. The exploitation of vinification by-products as feed additives appears to be a promising strategy to improve waste valorization and supply animals with bioactive molecules capable of improving animals' oxidative status and products' oxidative stability. [ABSTRACT FROM AUTHOR]

Six yeast strains belonging to Rhodosporidium toruloides, Lipomyces starkeyi, Rhodotorula glutinis and Cryptococcus curvatus were shake-flask cultured on xylose (initial sugar—S0 = 70 ± 10 g/L) under nitrogen-limited conditions. C. curvatus ATCC 20509 and L. starkeyi DSM 70296 were further cultured in media where process waters were partially replaced by the phenol-containing olive mill wastewaters (OMWs). In flasks with S0 ≈ 100 g/L and OMWs added yielding to initial phenolic compounds concentration (PCC0) between 0.0 g/L (blank experiment) and 2.0 g/L, C. curvatus presented maximum total dry cell weight—TDCWmax ≈ 27 g/L, in all cases. The more the PCC0 increased, the fewer lipids were produced. In OMW-enriched media with PCC0 ≈ 1.2 g/L, TDCW = 20.9 g/L containing ≈ 40% w/w of lipids was recorded. In L. starkeyi cultures, when PCC0 ≈ 2.0 g/L, TDCW ≈ 25 g/L was synthesized, whereas lipids in TDCW = 24–28% w/w, similar to the experiments without OMWs, were recorded. Non-negligible dephenolization and species-dependent decolorization of the wastewater occurred. A batch-bioreactor trial by C. curvatus only with xylose (S0 ≈ 110 g/L) was performed and TDCW = 35.1 g/L (lipids in TDCW = 44.3% w/w) was produced. Yeast total lipids were composed of oleic and palmitic and to lesser extent linoleic and stearic acids. C. curvatus lipids were mainly composed of nonpolar fractions (i.e., triacylglycerols). [ABSTRACT FROM AUTHOR]

One of the major environmental problems is the highly toxic agro-industrial waste called olive mill wastewater (OMW), deriving from olive oil production. On the other hand, the continuous development of the biological liquid fuel industry (biodiesel and bioethanol) makes it mandatory the process and exploitation of their main by-products, crude glycerol. This study dealt with the biotechnological conversions of biodiesel-derived crude glycerol with the use of the non-conventional yeast Yarrowia lipolytica in media that had been diluted with OMWs. OMWs, employed as simultaneous liquid medium and substrate, is a new trend recently appearing in Industrial Biotechnology, where value-added metabolites could be produced with simultaneous partial detoxification (i.e. decolorization and phenol removal) of the used residue. In the present study, diluted OMWs (containing 2.0 g/L of total phenolic compounds) blended with 70.0 g/L crude glycerol were employed as substrates. Production of value-added compounds by Y. lipolytica strain ACA-YC 5031 was studied in nitrogen-limited media favoring the production of secondary metabolites (i.e. citric acid, polyols, microbial lipids, polysaccharides). Batch-flask cultures were carried out and the impact of the addition of different NaCl concentrations (1.0%, 3.0%, 5.0% w/w) added upon the biochemical behavior of the strain was studied. Remarkable biomass production was observed in all trials, while in the "blank" experiment (no OMWs and no salt added), the metabolism was shifted toward the synthesis of polyols (Σpolyols = mannitol + arabitol + erythritol > 20 g/L and maximum total citric acid-Cit (sum of citric and isocitric acid) = 10.5 g/L). Addition of OMWs resulted in Citmax = 32.7 g/L, while Σpolyols concentration dropped to <15 g/L. Addition of salt in the OMW-based media slightly reduced the produced biomass, while Cit production drastically increased, reaching a final value of 54.0 g/L (conversion yield of Cit produced per unit of glycerol consumed = 0.82 g/g) in the trial with addition of 5.0% NaCl. Finally, significant color and phenols removal were observed, evaluating the yeast as a decontamination medium for the OMW and a great candidate for the production of value-added compounds. [ABSTRACT FROM AUTHOR]

Diversified mixed confectionery waste streams were utilized in a two-stage bioprocess to formulate a nutrient-rich fermentation media for microbial oil production. Solid-state fermentation was conducted for the production of crude enzyme consortia to be subsequently applied in hydrolytic reactions to break down starch, disaccharides, and proteins into monosaccharides, amino acids, and peptides. Crude hydrolysates were evaluated in bioconversion processes using the red yeast Rhodosporidium toruloides DSM 4444 both in batch and fed-batch mode. Under nitrogen-limiting conditions, during fed-batch cultures, the concentration of microbial lipids reached 16.6–17 g·L−1 with the intracellular content being more than 40% (w/w) in both hydrolysates applied. R. toruloides was able to metabolize mixed carbon sources without catabolite repression. The fatty acid profile of the produced lipids was altered based on the substrate employed in the bioconversion process. Microbial lipids were rich in polyunsaturated fatty acids, with oleic acid being the major fatty acid (61.7%, w/w). This study showed that mixed food side-streams could be valorized for the production of microbial oil with high unsaturation degree, pointing towards the potential to produce tailor-made lipids for specific food applications. Likewise, the proposed process conforms unequivocally to the principles of the circular economy, as the entire quantity of confectionery by-products are implemented to generate added-value compounds that will find applications in the same original industry, thus closing the loop. [ABSTRACT FROM AUTHOR]

Olive mill wastewaters (OMW) are the major effluent deriving from olive oil production and are considered as one of the most challenging agro-industrial wastes to treat. Crude glycerol is the main by-product of alcoholic beverage and oleochemical production activities including biodiesel production. The tremendous quantities of glycerol produced worldwide represent a serious environmental challenge. The aim of this study was to assess the ability of Yarrowia lipolytica strain ACA-DC 5029 to grow on nitrogen-limited submerged shake-flask cultures, in crude glycerol and OMW blends as well as in media with high initial glycerol concentration and produce biomass, cellular lipids, citric acid and polyols. The rationale of using such blends was the dilution of concentrated glycerol by OMW to (partially or fully) replace process tap water with a wastewater stream. The strain presented satisfactory growth in blends; citric acid production was not affected by OMW addition (Citmax~37.0 g/L, YCit/Glol~0.55 g/g) and microbial oil accumulation raised proportionally to OMW addition (Lmax~2.0 g/L, YL/X~20% w/w). Partial removal of color (~30%) and phenolic compounds (~10% w/w) of the blended media occurred. In media with high glycerol concentration, a shift towards erythritol production was noted (Erymax~66.0 g/L, YEry/Glol~0.39 g/g) simultaneously with high amounts of produced citric acid (Citmax~79.0 g/L, YCit/Glol~0.46 g/g). Fatty acid analysis of microbial lipids demonstrated that OMW addition in blended media and in excess carbon media with high glycerol concentration favored oleic acid production. [ABSTRACT FROM AUTHOR]

Biobased pigments are environmentally friendly alternatives to synthetic variants with an increased market demand. Production of pigments via fermentation is a promising process, yet optimization of the production yield and rate is crucial. Herein, we evaluated the potential of Penicillium purpurogenum to produce biobased pigments. Optimum sugar concentration was 30 g/L and optimum C:N ratio was 36:1 resulting in the production of 4.1-4.5 AU (namely Pigment Complex A). Supplementation with ammonium nitrate resulted in the production of 4.1-4.9 AU (namely Pigment Complex B). Pigments showed excellent pH stability. The major biopigments in Pigment Complex A were N-threonyl-rubropunctamin or the acid form of PP-R (red pigment), N-GABA-PP-V (violet pigment), PP-O (orange pigment) and monascorubrin . In Pigment Complex B, a novel biopigment annotated as N-GLA-PP-V was identified. Its basic structure contains a polyketide azaphilone with the same carboxyl-monascorubramine base structure as PP-V (violet pigment) and γ-carboxyglutamic acid (GLA) . The pigments were not cytotoxic up to 250 μg/mL.