Effect of Maillard reaction conditions on browning and antiradical activity of sugar–tuna stomach hydrolysate model system

The antiradical activity of Maillard reaction products (MRPs) made from sugar–tuna stomach hydrolysate model system was tested. The antiradical activity of the MRPs derived from ribose was 11-fold higher than that of MRPs derived from glucose due to the acyclic form of the ribose. The activity reached the plateau at a 30 mg/mL ribose concentration. The ribose caramelization contributed to the antiradical activity and browning reactions at 95 °C and 115 °C. The increase in DPPH radical scavenging of MRPs is attributed not only to the temperature but also to the buffer type and buffer concentration. Phosphate buffer showed the most efficient compared to citrate or Tris–HCl buffers. A positive correlation (R² = 0.98) was observed between the antiradical activity, the browning and the phosphate concentration. The MRPs obtained under these mild experimental conditions exhibited no toxicity towards Vero cells and 3T3 cells, despite their high antiradical activity.     

Phytosterols and tocopherols content of pulps and nuts of Brazilian fruits

Fruits and nuts from the North and Northeast regions of Brazil were collected to determine their phytosterol and tocopherol content. The species studied were Cotia nut (Aptandra spruceana M.), Brazil nut (Bertholletia excelsa H.B.K.), Mucajá (Couma rigida M.), Red Açaí (Euterpe oleracea M.), Inajá (Maximiliana maripa D.), Jenipapo (Genipa Americana L.), Buriti (Mauritia flexuosa L.) and Uxi (Endopleura uchi C.). Phytosterols were analyzed by GC–FID using β-cholestanol as an internal standard, while tocopherols were determined by RP-HPLC-DAD. The pulps of Mucajá (26–236 mg 100 g^–1), Inajá (119–285 mg 100 g^–1) and Jenipapo (216 mg 100 g^–1) showed the highest total phytosterol contents. Considering α-tocopherol equivalents, the pulps of Buriti (346.72 μg g^–1) and Uxi (200.92 μg g^–1) contained the highest vitamin E activity. Therefore, the results indicate that these fruits and nuts have great potential to be cultivated and marketed as alternative dietary sources for these bioactive compounds.     

Optimization of the associative growth of novel yoghurt cultures in the production of biomass, β-galactosidase and lactic acid using response surface methodology

The associative growth of Streptococcus thermophilus 95/2 (St 95/2) and Lactobacillus delbrueckii ssp. bulgaricus 77 (Lb 77) isolated from the Toros mountain region of Turkey was investigated with respect to lactic acid, biomass and β-galactosidase enzyme production using response surface methodology (RSM). The ratio (St 95/2:Lb 77) of the strains and media formulation had significant effect on all responses (p < 0.001). The predicted enzyme activity (2.14 U mL^?1), lactic acid (22.50 g L^?1) and biomass (7.11 g L^?1) production at optimum conditions were very close to the actual experimental values (2.14 U mL^?1, 22.94 g L^?1 and 7.86 g L^?1, respectively). The optimum conditions were to use these cultures in a ratio of 1.66:1.62 (St 95/2:Lb 77) in a medium containing whey (5%), corn steep liquor (4%), potassium phosphate (2%) and peptone (2%) at 43 °C for 8 h. The associative growth provided 6.4% and 39% more β-galactosidase activity and 8.73% and 44% more lactic acid compared with the results obtained using pure St 95/2 and Lb 77 strains, respectively.     

Dual-enzymatic synthesis of lactulose in organic-aqueous two-phase media

Lactulose has been successfully synthesized by dual-enzymatic method in organic-aqueous two-phase media using lactose and fructose as the raw materials. Cyclohexane–buffer system C₆H₁₂:buffer = 95:5 (v/v) was employed as the organic-aqueous media for the reaction. The dual-enzymatic system was consisted of immobilized lactase (IL) and immobilized glucose isomerase (IGI). Immobilized lactase was prepared by cross-linking the free lactase into Fe₃O₄-chitosan magnetic microspheres. The main enzymatic reaction parameters were investigated, including reaction temperature (T), pH value and reaction time (t). Under the optimum reaction conditions, i.e., lactose 0.8 g mL^?1, fructose 0.1 g mL^?1, IL 0.1 g mL^?1, IGI 0.05 g mL^?1, T = 30 °C, pH = 8.0 and t = 2 h, the obtained highest lactulose yield was approximately 151 g L^?1 and the corresponding productivity was 75.5 g L^?1 h^?1. Experimental results indicated that the organic-aqueous media can significantly promoted the transglycosidation activity of lactase and therefore improve the lactulose yield. The possible reaction mechanism of the synthesis of lactulose using IL and IGI in two-phase system was also proposed.     

Determination of moisture,solids-not-fat and fat-by-difference in butter using routine methods according to ISO 8851/IDF 191—an international collaborative study and a meta-analysis

An international collaborative study of routine methods for the determination of moisture, solids-not-fat (SNF) and fat-by-difference (i.e. fat (g 100 g^–1 product) =100−moisture (g 100 g^–1 product)−SNF (g 100 g^–1 product)) in butter was carried out to obtain estimates of the precision parameters repeatability (2.8s_r) and reproducibility (2.8s_R). Ten laboratories tested blind duplicate samples of eight different butters according to standard operating procedures jointly drafted by the International Standards Organisation (ISO) and the International Dairy Federation (IDF) (draft ISO standard 8851, parts 1–3; draft IDF standard 191, parts 1–3). Estimates of repeatability and reproducibility respectively were: moisture, 0.28 g 100 g^–1 product and 0.44 g 100 g^–1 product; SNF, 0.20 g 100 g^–1 product and 0.34 g 100 g^–1 product; fat-by-difference, 0.30 g 100 g^–1 product and 0.51 g 100 g^–1 product. These estimates are similar to those found previously in a New Zealand study. A meta-analysis, pooling the results of the present study and the New Zealand study, yielded overall estimates of repeatability and reproducibility as follows: moisture, 0.31 g 100 g^–1 product and 0.42 g 100 g^–1 product; SNF, 0.20 g 100 g^–1 product and 0.39 g 100 g^–1 product; fat-by-difference, 0.35 g 100 g^–1 product and 0.54 g 100 g^–1 product, and these are recommended for adoption in the respective methods. This study completes the validation work for the respective methods, which may now be progressed for final adoption as international standard routine methods. A protocol is proposed for dealing with situations where multiple precision estimates are available.     

Effect of commercial adjunct cultures on proteolysis in low-fat Kefalograviera-type cheese

The effect of two commercially available adjunct cultures, LBC 80 (Lactobacillus casei subsp. rhamnosus) and CR-213 (containing Lactococcus lactis subsp. cremoris and Lc. lactis subsp. lactis) on the proteolysis in low-fat hard ewes’ milk cheese of Kefalograviera-type was investigated. Two controls, a full-fat cheese (306 g kg⁻¹ fat, 378 g kg⁻¹ moisture) and a low-fat cheese (97 g kg⁻¹ fat, 486 g kg⁻¹ moisture, made using a modified procedure), were also prepared. The effect of adjunct culture on proteolysis, as examined by polyacrylamide gel electrophoresis of cheese and water soluble cheese extracts, was marginal. The reverse-phase HPLC peptide profiles of the water soluble extracts from low-fat cheeses were similar although some quantitative differences were observed between low-fat control cheese and experimental cheeses. The fat content as reflected by the differences in peptide profiles affected the pattern of proteolysis. Proteolysis, as measured by the percentage of total nitrogen soluble in water or in 120 g L⁻¹ trichloroacetic acid, was significantly (P<0.05) affected by the addition of adjunct cultures. Furthermore, the adjunct cultures enhanced the production of low molecular mass nitrogenous compounds; the levels of total nitrogen, soluble in 50 g L⁻¹ phosphotungstic acid, and of free amino acids were significantly (P<0.05) higher in the low-fat experimental cheeses than in the low-fat control cheese.     

Development of multiresidue DLLME and QuEChERS based LC–MS/MS method for determination of selected neonicotinoid insecticides in honey liqueur

Honey liqueur is an alcoholic drink derived from honey and strong fruit brandy of suitable type, traditionally made in Serbia and other Balkan countries. Although European Union has regulated the levels of neonicotinoid insecticides in honey and pollen, there is an increased risk of the presence of these compounds in traditional products made from honey. The objective of this study was to develop an optimized LC–MS/MS analytical method with dispersive liquid–liquid microextraction (DLLME) and QuEChERS sample preparation procedures for analysis of seven neonicotinoids (dinotefuran, nitenpyram, thiamethoxam, clothianidin, imidacloprid, acetamiprid and thiacloprid) in honey liqueur. The LC–MS/MS conditions were optimized to unequivocally provide good chromatographic separation, selectivity and specificity of developed method. The method was validated to fulfill the requirements of SANCO/12495/2011 for both sample pretreatment procedures providing results for accuracy (R, 69.2–113.4% for DLLME; 71.8–94.9% for QuEChERS), precision (RSD expressed in terms of repeatability (3.21–10.20% for DLLME; 4.19–12.81% for QuEChERS) and within-laboratory reproducibility (9.11–16.63% for DLLME; 11.32–16.40% for QuEChERS)), limits of detection (LOD, 0.5–1.5 μg L^− 1 for DLLME; 1.0–2.5 μg L^− 1 for QuEChERS) and quantification (LOQ, 1.0–5.0 μg L^− 1 for DLLME; 2.5–10.0 μg L^− 1 for QuEChERS). Matrix effects were compensated by the use of matrix-matched calibration. Analysis of real honey liqueur samples obtained from local markets showed the presence of clothianidin or thiacloprid in four of the analyzed samples, therefore implicating the necessity of ongoing control of this type of traditional product.     

Development and validation of a LC-ESI-MS/MS method for the determination of phenolic compounds in honeydew honeys with the diluted-and-shoot approach

A simple, reproducible and sensitive method has been optimized and validated for simultaneous determination of 32 phenolic compounds in bracatinga (Mimosa scabrella Benth.) with the diluted-and-shoot approach, without the need of any additional clean-up steps. It has been based on high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry and electrospray ionization (HPLC-ESI-MS/MS). The chromatography conditions were optimized, and due to the selectivity provided by MRM monitoring, LC separation required only 9 min. The developed method was validated on the basis of Eurachem and European Commission Decision 2002/657/EC guidelines. Mean recoveries ranged from 70.4 to 110%. Intra-day and inter-day precision with RSD (relative standard deviations) from 0.14 to 18.9% and 0.34 to 20.0%, respectively were achieved. Limits of detection (LOD) and quantification (LOQ) ranged from 0.03 to 3.20 μg L^− 1 and 0.20–12.8 μg L^− 1. Finally, the method was applied to samples and 20 phenolic compounds were quantified in all the samples analyzed, representing a contribution to the characterization and quantification of phenolic compounds from bracatinga (M. scabrella Bentham) honeydew honey.     

In-vitro mineral binding capacity of five fiber sources and their insoluble components for magnesium and calcium1

In-vitro binding of calcium (Ca) and magnesium (Mg) by total dietary fiber, hemicellulose A (HCL A), lignocellulose (LCL), cellulose (CL), and lignin (L) fractions isolated from rice bran (RB), wheat bran (WB), oat fiber (OF), apple fiber (AF) and tomato fiber (TF) was evaluated. At pH 6.8, significant amounts of Ca were bound by whole fibers, ranging from 800 μg g⁻¹ for RB to 10 097 μg g⁻¹ for TF. Mg bound by whole fibers varied from 496 μg g⁻¹ for OF to 2177 μg g⁻¹ for WB. Re-acid washing (pH<2.0) released 95–99% of the Ca and Mg bound to the fibers. Fibers with the highest endogenous Ca and Mg concentrations bound significantly (P<0.05) the highest amounts of the minerals studied. The Ca bound by HCL A varied from 9753 μg g⁻¹ for RB to 11 337 mg g⁻¹ for TF, whereas Mg bound varied from 1151 μg g⁻¹ for OF to 5626  μg g⁻¹ for TF hemicellulose fractions, respectively. Among the fiber components, Mg binding decreased in the order HCL A>LCL>L>CL, whereas Ca bound was in the order HCL A>LCL>CL>L. A relatively strong correlation was observed between the combined effects of protein content, hemicellulose, and lignin vs total Ca and Mg bound. 1998 Elsevier Science Ltd. All rights reserved     

Effective valorisation of distillery stillage by integrated production of lactic acid and high quality feed

Utilization of distillery stillage from bioethanol production for lactic acid and feed production was studied. The lactic acid fermentation of the stillage was performed by Lactobacillus rhamnosus ATCC 7469 and maximal lactic acid concentration of 50.18 g L^− 1, yield of 0.90 g g^− 1, productivity of 1.48 g L^− 1 h^− 1 and viable cell number of 5 × 10⁹ CFU mL^− 1 were achieved. Solid residues with biomass remains after lactic acid fermentation were assessed for animal consumption. The content of proteins and ash decreased in the residues after the fermentation, whilst the content of oil and nitrogen free extract was higher when compared to unfermented samples. The digestible (17480.64 kJ kg^− 1) and metabolisable (17389.08 kJ kg^− 1) energies as well as digestibility (966.95 g kg^− 1) of the fermentation residue were very high. The in vitro assessment of L. rhamnosus ATCC 7469 survival in simulated gastric conditions has shown high survival rate (87%). In addition, this bacterium has shown good antimicrobial activity against the most important pathogens and capability to produce exopolysaccharide on different sugars present in animal diet. After effective lactic acid fermentation, the residues could be recommended as a high quality feed for monogastric animals.     

Synthesis of potentially-bioactive lactosyl-oligofructosides by a novel bi-enzymatic system using bacterial fructansucrases

Efficient enzymatic synthesis of lactosyl-oligofructosides (LFOS) with a degree of polymerization from 4 to 8 was achieved in the presence of sucrose:lactosucrose and sucrose:lactose mixtures by transfructosylation reaction. The main synthesized LFOS which consist of β-2,1-linked fructose to lactosucrose: β-d-galactopyranosyl-(1 → 4)-α-d-glucopyranosyl-[(1 → 2)-β-d-fructofuranosyl]n-(1 → 2)-β-d-fructofuranoside (where n refers to the number of transferred fructose moieties) was structurally characterized by nuclear magnetic resonance (NMR). The maximum formation of LFOS was 81% (in weight with respect to the initial amount of lactosucrose) and was obtained after 24 h of transfructosylation reaction based on sucrose:lactosucrose (250 g L^− 1 each) catalyzed by an inulosucrase from Lactobacillus gasseri DSM 20604 (IS). The production of LFOS in the presence of sucrose:lactose mixtures required a previous high-yield lactosucrose synthesis step catalyzed by using a levansucrase from Bacillus subtilis CECT 39 (LS) before the inulosucrase-catalyzed reaction. This novel one-pot bi-enzymatic system led to the synthesis of about 22% LFOS in weight, with respect to the initial amount of lactose (250 g L^− 1). The results revealed a high specificity for the substrate involved in the inulosucrase-catalyzed reaction given that, although lactosucrose (O-β-d-galactopyranosyl-(1 → 4)-O-α-d-glucopyranosyl-(1 → 2)-β-d-fructofuranoside) acted as a strong acceptor of β-2,1-linked fructose, lactose (β-d-galactopyranosyl-(1 → 4)-α-d-glucose) was found to be an extremely weak acceptor.     

Routine and sensitive SPE-HPLC method for quantitative determination of pheophytin a and pyropheophytin a in olive oils

A sensitive and specific HPLC method with tandem diode array-fluorescence detection (DAD-FL) has been developed and validated for the simultaneous determination of pheophytin a (phy a) and pyropheophytin a (pyrophy a) in olive oils. Pigments were extracted with reverse phase solid phase extraction (RP-SPE) and subsequently analysed by HPLC-DAD-FL. The chromatographic analysis was carried out isocratically on ODS2 RP column using methanol–acetone (1:1 v/v) at flow-rate of 2.0 ml min⁻¹. Specificity of the method was assured by the simultaneous detection by UV–visible (410 nm) and FL (λ_Ex: 410 nm; λ_Em: 672 nm). Both compounds could be baseline separated within 7 min. The method was validated and applied in olive oil samples recently extracted as well as stored during 12 months. The limit of detection (LOD) defined at a signal-to-noise ratio of about 3 was ∼21.6 ng g⁻¹ for pyrophy a and ∼24.6 ng g⁻¹ for phy a under FL detection, and ∼148.0 ng g⁻¹ for both analytes under UV–visible detection. The calibration graphs were linear (r² > 0.9999; p < 0.01) between 0.25–14.00 ng μl⁻¹ for pyrophy a and 0.25–19.00 ng μl⁻¹ for phy a, under both fluorescence and UV–visible detection conditions. Recoveries of phy a and pyrophy a were over 94% as estimated by the standard addition method. Relative standard deviation for the intra-day and inter-day determination of phy a and pyrophy a were lower than 3.7% and 8.0%, respectively.     

Effect of combinations of high-pressure treatment and bacteriocin-producing lactic acid bacteria on the survival of Listeria monocytogenes in raw milk cheese

The combined effect of high-pressure (HP) treatment and bacteriocin-producing lactic acid bacteria (BP-LAB) on the survival of Listeria monocytogenes Scott A in cheeses made from raw milk that was inoculated with the pathogen at 4.80 log cfu mL⁻¹, a commercial starter and one of seven strains of BP-LAB was investigated. On day 3, the counts of L. monocytogenes were 7.03 log cfu g⁻¹ in a control cheese (without BP-LAB, not HP treated), 6.06–6.74 log cfu g⁻¹ in cheeses with BP-LAB, 6.13 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 300 MPa, 2.01 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 500 MPa, 3.83–5.43 log cfu g⁻¹ in cheeses with BP-LAB and treated on day 2 at 300 MPa, and 1.81 log cfu g⁻¹ or less in cheeses with BP-LAB and treated on day 2 at 500 MPa. HP treatment was more effective on day 51 than on day 2.     

Incorporation of microbial transglutaminase into non-fat yogurt production

This study investigated the physical, chemical and sensory characteristics of non-fat yogurts treated with microbial transglutaminase (MTGase) at varying concentrations from 0 to 0.5 g L⁻¹. Also, the effect of enzyme inactivation prior to fermentation on the selected properties of the yogurts was studied. Acid development rate was reduced with increasing MTGase doses. Cross-linking of milk proteins by MTGase had a growth-slowing effect on yogurt starter bacteria, which was more pronounced at higher concentrations. Physical properties of the yogurts were improved by MTGase throughout 21-day storage; on the contrary, the production of acetaldehyde was slowed down by increasing MTGase concentrations during the same period. Principal component analysis (PCA) and hierarchial cluster analysis (HCA) clearly differentiated the samples with added MTGase at lower (⩽0.3 g L⁻¹) and higher (0.4–0.5 g L⁻¹) concentrations regarding the physical and sensory properties. The physical and sensory properties of non-fat set yogurt could be improved by incorporating MTGase up to a level of 0.3 g L⁻¹.     

Isolation and characterisation of a novel antibacterial peptide from bovine αS1-casein

Bovine casein was hydrolysed with a range of proteolytic enzymes including pepsin, trypsin, α-chymotrypsin and β-chymotrypsin, and assessed for antibacterial activity. The pepsin digest of bovine casein, which showed antibacterial activity, was fractionated using reverse phase high performance liquid chromatography and the antibacterial peptides isolated were characterised using electrospray ionisation mass spectrometry. Two antibacterial peptides were identified, a novel peptide (Cp1) which corresponded to residues 99–109 of bovine α_S1-casein and a previously reported peptide (Cp2) which corresponded to residues 183–207 of bovine α_S2-casein. The minimum inhibitory concentration (MIC) of Cp1 and Cp2 were determined against a range of bacterial cultures. Cp1 exhibited an MIC of 125 μg mL⁻¹ against all Gram-positive bacteria tested, and MIC ranging between 125 and >1000 μg mL⁻¹ against the Gram-negative bacteria tested. Cp2 was generally far more potent against the Gram-positive bacteria, exhibiting an MIC of 21 μg mL⁻¹, compared to MICs ranging from 332 to >664 μg mL⁻¹ against most of the Gram-negative bacteria tested.     

Influence of methylcellulose on dynamic rheology of dilute wheat gliadin solution in 50% (v/v) aqueous propanol

Influences of methylcellulose on dynamic rheological behaviors of 50 g L⁻¹ (dilute) wheat gliadins in 50% (v/v) aqueous propanol were investigated as a function of methylcellulose concentration C ranging from 5 to 20 g L⁻¹. The mixture solutions exhibit shear rate thinning that becomes pronounced with increasing C. Specific viscosities of the first and the second Newtonian plateau viscosities, as well as the steady-flow viscosity as a function of C, reveal that methylcellulose is in the semi-dilute unentangled regime at temperatures from 5 to 65 °C. The intermolecular interaction between methylcellulose and gliadins provides high flow resistance, improved shear thinning and steady-flow activation energy of the mixture solution in comparison with pure gliadins solution.     

Wine lees valorization: Biorefinery development including production of a generic fermentation feedstock employed for poly(3-hydroxybutyrate) synthesis

This study demonstrates the development of a novel wine lees (WL) based integrated biorefinery for the production of several added-value products. WL were initially fractionated for the production of antioxidants, tartrate and ethanol and the remaining stream was converted into a fermentation nutrient supplement for poly(3-hydroxybutyrate) (PHB) production using the strain Cupriavidus necator DSM 7237. Hydrolysis of pretreated WL was carried out using crude enzyme consortia produced via solid state fermentation of Aspergillus oryzae. Optimization of hydrolysis was based on the enhancement of total Kjeldahl nitrogen to free amino nitrogen (FAN) conversion yield by evaluating the effect of the initial pH value, temperature, initial proteolytic activity and initial WL concentration. WL hydrolysates and crude glycerol were used as nutrient and carbon sources, respectively, in batch and fed-batch fermentations for the production of PHB. Bacterial growth and PHB production were influenced significantly by the FAN content of the WL derived hydrolysates and by the addition of trace elements. Using an initial FAN concentration of 700 mg L^− 1 and supplementation with trace elements led to the production of 30.1 g L^− 1 of PHB concentration with an intracellular content of 71.3% (w w^-1) and a productivity of 0.56 g L^− 1 h^− 1 during fed-batch fermentation.     

Available lysine,protein digestibility and lactulose in commercial infant formulas

Available lysine, in vitro protein digestibility and lactulose values were determined in 23 commercial infant formulas. The mean available lysine content of the formulas based on dairy proteins was 66.7±9.5 mg g⁻¹ protein, similar to that of human milk, while that of soy based formulas was considerably lower (45.0±8.3 mg g⁻¹ protein). In vitro protein digestibility values ranged 85.5–88.9% for soy-based formulas and 90.5–98.3% for formulas based on dairy proteins. Formulas based on milk enriched with whey had higher lactulose content than those based on cow's milk. However, all values were below the limit of 600 mg L⁻¹ recommended for UHT milk.     

Production of volatile aroma compounds by kefir starter cultures

Production of carbonyl compounds by single-strain cultures, kefir starter (Lactobacillus delbrueckii subsp. bulgaricus HP1+Lb. helveticus MP12+Lactococcus lactis subsp. lactis C15+Streptococcus thermophilus T15+Saccharomyces cerevisiae A13) and kefir grains during fermentation and storage of kefir was studied. The content of carbonyl compounds produced by kefir starter was greater than that produced by kefir grains. The maximum acetaldehyde concentration (18.3 μg g⁻¹) in kefir with starter culture was mainly due to the metabolic activity of Lb. delbrueckii subsp. bulgaricus HP1 isolated from kefir grains. The highest diacetyl production activity was recorded in the starter culture (1.87 μg g⁻¹) and the single-strain culture St. thermophilus T15 (1.62 μg g⁻¹), followed by Lb. helveticus MP12 (0.85 μg g⁻¹) and Lc. lactis subsp. lactis C15 (0.42 μg g⁻¹). The lactobacilli Lb. delbrueckii subsp. bulgaricus HP1 and Lb. helveticus MP12 produced acetone, which was not found in the cocci cultures. The presence of 2-butanone was related to the production ability of Lb. helveticus MP12. In comparison, Lc. lactis subsp. lactis C15 synthesized ethyl acetate more actively than the other single-strain cultures included in the starter. S. cerevisiae A13 produced ethanol and CO₂ in amounts (3975 μg g⁻¹; 1.80 g L⁻¹) that lent cultured kefir distinctive flavour and aroma characteristic of authentic kefir.