Augmenting Functional and Sensorial Quality Attributes of Kefir through Fortification with Encapsulated Blackberry Juice.

Kefir is a fermented dairy product claimed to confer many health-promoting effects, but its acidic taste is not appealing to some consumers. Therefore, the aim of this study was to enhance the functional and sensorial quality attributes of kefir through fortification with encapsulated blackberry juice (EBJ). The blackberry juice was successfully encapsulated via freeze-drying using lentil protein isolate (LPI) as the carrier. The encapsulated blackberry juice showed good physicochemical, functional, and morphological properties, as well as microbiological safety for use as a food additive. The kefir was fortified with EBJ in concentrations of 1, 2.5, 5, and 7.5% (w/w), stored for up to 28 days under refrigeration, and periodically evaluated. Parameters such as the viscosity, titrable acidity, and pH indicate that the kefir fortification did not affect its stability during storage. EBJ significantly increased the antioxidant properties of the kefir, depending on the fortification level. Additionally, all the fortified samples provided more anthocyanins than the daily recommended intake. Microbiological profiling demonstrated that good laboratory practice and hygiene were implemented during the experiments. Finally, the panelists showed that higher EBJ concentrations in the kefir resulted in greater overall acceptability, indicating that this encapsulate has the potential to be a substitute synthetic color additive in the dairy industry.

Integration of enzymatic modification and ultrafiltration for the production of pectin fractions with highly potent antioxidant capacity as green valorization of sugar beet pulp.

Enzymatic hydrolysis of pectin followed by ultrafiltration of hydrolysate was applied with the aim to produce fractions with potent antioxidant capacity. Pectin was isolated from waste sugar beet pulp by acidic extraction, washed by diafiltration and concentrated by ultrafiltration. Enzymatic hydrolysis was performed with endo-polygalacturonase, and hydrolysate was processed by ultrafiltration into four fractions using membranes in series of decreasing cut-offs from 10 to 1 kDa. Hydrolysis with endo-polygalacturonase increased total antioxidant capacity by twofold in comparison to un-hydrolyzed pectin. Antioxidant capacity of all fractions was considerably higher than that of pectin-from 14.7 to 25-fold, for fraction containing fragments 10 kDa > Mw > 5 kDa and Mw < 1 kDa, respectively. Considerable increase of total antioxidant capacity of pectin through the integration of enzymatic modification and ultrafiltration fractionation indicated great potential of applied green protocol for the production of high-value hydrolysates of pectin from waste sugar beet pulp.

Valorisation of Lignocellulosic Wastes, the Case Study of Eucalypt Stumps Lignin as Bioadsorbent for the Removal of Cr(VI).

The main objective of this work was to assess Eucalyptus globulus lignin as an adsorbent and compare the results with kraft lignin, which has previously been demonstrated to be an effective adsorbent. Eucalypt lignin was extracted (by the dioxane technique), characterised, and its adsorption properties for Cr(VI) ions were evaluated. The monomeric composition of both types of lignin indicated a high content of guaiacyl (G) and syringyl (S) units but low content of p-hydroxyphenyl (H), with an H:G:S ratio of 1:50:146 (eucalypt lignin) and 1:16:26 (kraft lignin), as determined by Py-GC/MS. According to elemental analysis, sulphur (2%) and sodium (1%) were found in kraft lignin, but not in eucalypt lignin. The adsorption capacity of the eucalypt lignin was notably higher than the kraft lignin during the first 8 h, but practically all the ions had been absorbed by both the eucalypt and kraft lignin after 24 h (93.4% and 95%, respectively). Cr(VI) adsorption onto both lignins fitted well using the Langmuir adsorption isotherm model, with capacities of 256.4 and 303.0 mg/g, respectively, for eucalypt and kraft. The study's overall results demonstrate the great potential of eucalypt lignin as a biosorbent for Cr(VI) removal from aqueous solutions.

The effect of enzymatic pretreatment of chickpea on functional properties and antioxidant activity of alkaline protein isolate.

Functional properties and antioxidant activity of protein isolates extracted by alkali from chickpea seed after treatment with cellulase and xylanase cocktail and individual arabinofuranosidase were evaluated. Both enzymatic treatments improved recovery of protein by more than 30%, moreover, arabinofuranosidase enabled higher extraction efficiency - above 93%. Protein extracted after treatment with enzyme cocktail showed improved solubility and oil holding capacity by 14% and 80%, respectively, while water holding capacity was increased by 130% after both applied enzymatic pretreatments. The action of enzyme cocktail was more beneficial for improving emulsifying activity and stability of alkaline isolate by 22% and 31%, respectively. Whipping properties of alkaline protein isolate were significantly improved when both enzymatic pretreatments were applied with foam stability increased by 150%. Protein from extractions with arabinofuranosidase and enzyme cocktail expressed enhanced antioxidant activity by 70% and 110%, respectively. Analysis of protein pattern and structural characteristics indicated differences between investigated isolates.

Impact of Different Enzymatic Processes on Antioxidant, Nutritional and Functional Properties of Soy Protein Hydrolysates Incorporated into Novel Cookies.

Soy protein concentrate (SPC) was hydrolyzed using several commercial food-grade proteases (Alcalase, Neutrase, papain, Everlase, Umamizyme, Flavourzyme) and their combination to obtain promising ingredients in the manufacture of functional bakery products. In all cases, the hydrolysis caused nutritional, sensory, and rheological changes in SPC, as well as protein structural changes like increased surface hydrophobicity and content of exposed SH groups with the magnitude of these changes depending on enzyme specificity. The hydrolysis with the combination of Neutrase and Flavourzyme (NeuFlav) increased essential amino acid content by 9.8% and that of Lys by 32.6% compared to SPC. This hydrolysate showed also significant antioxidant activities including ABTS and superoxide anion scavenging activity and metal-chelating ability. The addition of all hydrolysates in wheat flour decreased water adsorption and increased development time to some extent due to gluten network weakening, but also decreased the rate of starch retrogradation, contributing to the increase of the shelf-life of bakery products. The NeuFlav tasted less bitter than other hydrolysates, while E-nose provided a discrimination index of 93 between control and hydrolysates. It appeared that the addition of the NeuFlav hydrolysate in a cookie formulation improved protein content and nutritional quality and directed to its higher general consumer acceptability than cookies formulated with only wheat flour.

Advance diversity of enzymatically modified arabinoxylan from wheat chaff.

Hydrolysates of arabinoxylan extracted from wheat chaff were prepared using different enzymatic treatments with an emphasis on improvements in their anti-diabetic, antioxidant and functional characteristics. The extracted arabinoxylan was subjected to enzymatic hydrolysis using individual xylanase, arabinofuranosidase, and feruloyl esterase, and their combinations. In all obtained hydrolysates, peaks corresponding to molecular weight lower than 38 kDa were noticed, while non-hydrolysed arabinoxylan had only peaks corresponding to 580 and 38 kDa. Results indicated that applied enzymes could hydrolyse polymeric arabinoxylan while their synergistic actions successfully modified its structure reflecting in lowered viscosity. Besides, it has been observed that the synergistic actions of enzymes improved the biological activities of arabinoxylan more than twice. Chemometric classification analysis showed that synergistic enzymes' actions were predominantly responsible for the improvement of biological activities. It indicated that they might be a useful tool for diversification and enhancement of biological activities of arabinoxylan from wheat chaff.

Environmental-friendly technologies for the production of antioxidant xylooligosaccharides from wheat chaff.

Ultrasound irradiation and enzymatic hydrolysis were applied for the production of antioxidant xylooligosaccharides from wheat chaff. To facilitate enzyme action, 3% (w/w) suspension of raw material was pretreated by ultrasound at 25Hz, 540W for 10min. Ultrasound treatment released xylooligosaccharides with predominant xylotriose into liquid fraction which expressed 1.03±0.01 (µmolAAE/g) ABTS radicals-scavenging activity. Endo-xylanase action on pretreated wheat chaff released 21.76±1.42 and 32.3±0.75mg reducing sugars equivalents/g after 24h when applied at dosages 0.15 and 0.3U/g. respectively. With increase in reaction time portion of xylotriose and xylotetraose in hydrolysates was increase relatively to xylopentose as well as quantities of xylose and glucose. Extremely significant inverse correlation between monosaccharides/oligosaccharides ratio and ABTS radical-scavenging activity was determined. Results indicated potential of environmental friendly ultrasound and enzymatic technologies for the production of xylooligosaccharides from wheat chaff for potential food application.

Covalent Immobilization of Enzymes on Eupergit® Supports: Effect of the Immobilization Protocol.

A selection of best combination of adequate immobilization support and efficient immobilization method is still a key requirement for successful application of immobilized enzymes on an industrial level. Eupergit® supports exhibit good mechanical and chemical properties and allow establishment of satisfactory hydrodynamic regime in enzyme reactors. This is advantageous for their wide application in enzyme immobilization after finding the most favorable immobilization method. Methods for enzyme immobilization that have been previously reported as efficient considering the obtained activity of immobilized enzyme are presented: direct binding to polymers via their epoxy groups, binding to polymers via a spacer made from ethylene diamine/glutaraldehyde, and coupling the periodate-oxidized sugar moieties of the enzymes to the polymer beads. The modification of the conventionally immobilized enzyme with ethylenediamine via the carbodiimide route seems to be a powerful tool to improve its stability and catalytic activity.

Synthesis of aliphatic esters of cinnamic acid as potential lipophilic antioxidants catalyzed by lipase B from Candida antarctica.

Immobilized lipase from Candida antarctica (Novozyme 435) was tested for the synthesis of various phenolic acid esters (ethyl and n-butyl cinnamate, ethyl p-coumarate and n-butyl p-methoxycinnamate). The second-order kinetic model was used to mathematically describe the reaction kinetics and to compare present processes quantitatively. It was found that the model agreed well with the experimental data. Further, the effect of alcohol type on the esterification of cinnamic acid was investigated. The immobilized lipase showed more ability to catalyze the synthesis of butyl cinnamate. Therefore, the process was optimized for the synthesis of butyl cinnamate as a function of solvent polarity (logP) and amount of biocatalyst. The highest ester yield of 60.7 % was obtained for the highest enzyme concentration tested (3 % w/w), but the productivity was for 34 % lower than the corresponding value obtained for the enzyme concentration of 1 % (w/w). The synthesized esters were purified, identified, and screened for antioxidant activities. Both DPPH assay and cyclic voltammetry measurement have shown that cinnamic acid esters have better antioxidant properties than cinnamic acid itself.

The immobilization of enzyme on Eupergit® supports by covalent attachment.

A selection of the best combination of adequate immobilization support and efficient immobilization method is still a key requirement for successful application of immobilized enzymes on the industrial level. Eupergit) supports exhibit good mechanical and chemical properties and allow establishment of satisfactory hydrodynamic regime in enzyme reactors. This is a good recommendation for their wide application in enzyme immobilization after finding the most favorable immobilization method. Methods for enzyme immobilization that have been previously reported as efficient, considering the obtained activity of immobilized enzyme are presented: direct binding to polymers via their epoxy groups, binding to polymers via a spacer made from ethylene diamine/glutaraldehyde, and coupling the periodate-oxidized sugar moieties of the enzymes to the polymer beads.

Proteins from common bean (Phaseolus vulgaris) seed as a natural coagulant for potential application in water turbidity removal.

The ability of coagulation active proteins from common bean (Phaseolus vulgaris) seed for the removal of water turbidity was studied. Partial purification of protein coagulant was performed by precipitation with ammonium sulphate, dialysis and anion exchange chromatography. Adsorption parameters for ion-exchange process were established using dialysate extract. Results revealed that the highest values of the adsorbed protein were achieved in 50 mmol/L phosphate buffer at pH 7.5 and the maximum adsorption capacity was calculated to be 0.51 mg protein/mL matrix. Partially purified coagulant at initial turbidity 35 NTU expressed the highest value of coagulation activity, 72.3%, which was almost 22 times higher than those obtained by crude extract considering applied dosages. At the same time, the increase in organic matter that remained in water after coagulation with purified protein coagulant was more than 16 times lower than those with crude extract, relatively to its content in blank.

Sequence polymorphism of the mitochondrial DNA control region in the population of Vojvodina Province, Serbia.

In order to generate and establish the database for forensic identification purposes in Vojvodina Province (Serbia), the sequence of the hypervariable regions 1 (HV1) and 2 (HV2) of the mtDNA control region were determined in a population of 104 unrelated individuals from Vojvodina Province, using a fluorescent-based capillary electrophoresis sequencing method. A total of 93 different haplotypes were found, of these 83 mtDNA types were unique, nine haplotypes were shared by two individuals and one haplotype by three individuals. The variation of mtDNA HV1 and HV2 regions was confined to 116 nucleotide positions, of which 72 were observed in the HV1 and 44 in the HV2. A statistical estimate of the results for this population showed the genetic diversity of 0.9977 and the random match probability of 1.18%. Haplogroup H was the most common haplogroup (43.3%). Haplogroups observed at intermediate levels included clusters U (13.5%), T (10.6%), J (8.6%) and W (5.8%).

Removal of water turbidity by natural coagulants obtained from chestnut and acorn.

The ability of seed extracts of several species of chestnut and acorn to act as natural coagulants was tested using a synthetic turbid water. Active components were extracted from ground seeds of Horse chestnut and acorns of some species of family Fagaceae: Common oak, Turkey oak, Northern red oak and European chestnut. All investigated extracts had coagulation capabilities and their amounts depended on pH values and initial turbidities. The seed extracts from European chestnut and Common oak acorn were the most efficient expressing the highest coagulation activities, about 80% and 70%, respectively, in both low and medium investigated water turbidities at the lowest coagulant dose 0.5 ml/L.

Pectinase partitioning in polyethylene glycol 1000/Na2SO4 aqueous two-phase system: statistical modeling of the experimental results.

Statistical models concerning partitioning of pectinase in polyethylene glycol 1000/Na2SO4 aqueous two-phase system were established with response surface methodology. Concentrations of polyethylene glycol 1000 and Na2SO4 were selected as independent variables to evaluate their impact on parameters of partitioning in aqueous two-phase system--the partition coefficient of pectinase, purification factor and pectinase yield. An experimental space where over 2.5-fold purification was achieved, followed by over 90% yield of pectinase. The established models showed good prediction of partitioning parameters.

Affinity partitioning of a Cellulomonas fimi beta-mannanase with a mannan-binding module in galactomannan/starch aqueous two-phase system.

A new approach in affinity separations was studied by partitioning of Cellulomonas fimi beta-mannanase (EC 3.2.1.78) containing a mannan-binding module in galactomannan/hydroxypropyl starch aqueous two-phase system. Comparison was made with a truncated version of C. fimi beta-mannanase which lacked the mannan-binding module. Results showed that affinity partitioning of the beta-mannanase was achieved due to biospecificity of the mannan-binding module towards the top phase containing galactomannan. Experiments were conducted at pH 8 to prevent enzyme degradation of the phase containing galactomannan. Removal of the top phase polymer was accomplished by beta-mannanase degradation allowed by shifting to the optimal pH 6. In the combination with the genetic fusion of any given protein to the mannan-binding module, the results envision a general procedure for primary affinity recovery of such fusion proteins.