Argentine Patagonia barberry chemical composition and evaluation of its antioxidant capacity.

An important portion of vitamins, minerals and polyphenols components in human diet are captured from fruit consumption. Argentinean Patagonia Berberis microphylla was characterized with the phenolic content, the proximate composition and the identification and quantification of anthocyanins, not-anthocyanins and proteins. The antioxidant capacity of berberis ethanolic extracts (EB) was determined by the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. EB was used to reduce production of reactive substances species (ROS) in zebrafish. EB presented a total polyphenols content of 1,035.03 mg GAE/100 g fresh weight (FW). EB presented an ABTS value of 116.25 ± 17 µmol TE/g FW. EB presented a DPPH value of 137.80 ± 1.90 µmol TE/g FW. EB was able of reducing the ROS in zebrafish. Berberies Protein Isolate (BPI) presented proteins with bands from 15 to 62 kDa. BPI presented an ABTS value of 593.11 ± 8.60 µmol TE/g. The BPI duodenal digest presented a value of 641.07 ± 12.60 µmol TE/g digests. PRACTICAL APPLICATIONS: The practical applications of the present study are to increase scientific knowledge for consumers about the quality and benefits of the consumption of the native fruit (Berberis microphylla) from the Patagonia region of Argentine. This work describes the protein profile of berberies, their digestibility and their antioxidant activity. This study allows to better understand the phytonutrients that make up this fruit. Future studies may identify the peptides present in hydrolyzates. The bio-compounds of this fruit could be used as functional ingredients by the food industry for different purposes.

Production of White, Red and Black Quinoa (Chenopodium quinoa Willd Var. Real) Protein Isolates and Its Hydrolysates in Germinated and Non-Germinated Quinoa Samples and Antioxidant Activity Evaluation.

Red, black and white seeds quinoa were germinated at 28 °C during 24 (G1), 48 and 72 h (G3). Red quinoa presented a higher percentage of germination with a value of 46% of germination at 72 h. Quinoa protein isolate (QPI) was obtained by alkaline extraction (pH 8.0) followed by an isoelectric precipitation (pH 4.5) from white, red and black quinoa seeds, germinated QPI-G1 or QPI-G3 and non-germinated QPI-NG, Chenopodium quinoa Willd var. Real. QPI-G1, QPI-G3 and QPI-NG were subject to a simulated gastric digestion (DG) and in vitro duodenal digestion (DD). The antioxidant activity was evaluated using the 1, 1-diphenyl-2-picryl hydrazyl (DPPH), azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and oxygen radical absorbance capacity (ORAC) methods. Gastric and duodenal digest of QPI-NG and QPI-G1 and QPI-G3 from white, red and black quinoa presented antioxidant activity. QPI-G1-DD of white quinoa presented the highest antioxidant activity with a DPPH value of 167.98 µmoL TE/g of digest, QPI-G1-DD of red quinoa with an ABTS value of 204.86 µmoL TE/g of digest and QPI-G1-DD of black quinoa with an ORAC value of 401.42 µmoL TE/g of digest. QPI-G3-DD of white quinoa presented higher antioxidant activity with a DPPH value of 186.28 µmoL TE/g of sample, QPI-G3-DD of red quinoa with an ABTS value of 144.06 µmoL TE/g of digest and QPI-G3-DD of black quinoa with an ORAC value of 395.14 µmoL TE/g of digest. The inhibition of reactive oxygen species (ROS) production in the zebrafish embryo model (Danio rerio) was evaluated. Protein profiles of QPI from white, red and black from germinated quinoa and non-germinated quinoa were similar with proteins between 10 kDa to 100 kDa with the presence of globulins 11S and 7S and 2S albumins.

Inhibition of Lipid Peroxidation of Kiwicha (Amaranthus caudatus) Hydrolyzed Protein Using Zebrafish Larvae and Embryos.

Amaranth protein concentrate (APC) was hydrolyzed under in vitro gastrointestinal conditions. APC proteins were partially degraded by pepsin at pHs 1.2, 2.0, and 3.2. During the intestinal phase (pepsin/pancreatin enzymes at pH 7.0), no polypeptide bands were observed in the gel, suggesting the susceptibility of amaranth proteins to the action of digestive enzymes. The potent in vitro inhibition of lipid peroxidation, shown by the gastric and intestinal digests, was confirmed in the zebrafish larvae, with a 72.86% reduction in oxidation of lipids in the presence of the gastric hydrolysate at pH 2.0, compared to a 95.72% reduction in the presence of the gastrointestinal digest. APC digests were capable of reducing reactive oxygen species (ROS) production in the zebrafish embryo model with a value of fluorescence of 52.5% for the gastric hydrolysate, and 48.4% for the intestinal hydrolysate.

Production of hesperetin using a covalently multipoint immobilized diglycosidase from Acremonium sp. DSM24697.

The diglycosidase α-rhamnosyl-ß-glucosidase (EC 3.2.1.168) from the fungus Acremonium sp. DSM24697 was immobilized on several agarose-based supports. Covalent multipoint immobilization onto glyoxyl-activated agarose was selected as the more stable preparation at high concentration of dimethyl sulfoxide (DMSO) and high temperature. The optimal conditions for the immobilization process involved an incubation of the enzyme with agarose beads containing 220 µmol of glyoxyl groups per gram at pH 10 and 25°C for 24 h. The hydrolysis of hesperidin carried out in 10% v/v DMSO at 60°C for 2 h reached 64.6% substrate conversion and a specific productivity of 2.40 mmol h(-1) g(-1). Under these conditions, the process was performed reutilizing the catalyst for up to 18 cycles, maintaining >80% of the initial activity and a constant productivity 2.96 ± 0.42 µmol(-1) h(-1) g(-1). To the best of our knowledge, such productivity is the highest achieved for hesperetin production through an enzymatic approach.

Polyvinyl alcohol-pectin cryogel films for controlled release of enrofloxacin.

The release of enrofloxacin entrapped in polyvinyl alcohol (PVA) cryogel at pH 5.5 showed a first-order kinetic, releasing 69.7% of the antibiotic after 4.5 h at 37 °C. In order to slow down the fluoroquinolone release rate, high-methoxylated pectin was added into the cryogel (PVA-P). A film containing 1.0% (w/v) HM pectin and 5.0 µg/ml enrofloxacin released only 3.7% of the antibiotic after 4.5 h. Since the FTIR spectrum showed that most of the interactions between PVA-P matrix and enrofloxacin were due to polar groups (carboxylate and amine), a two-layer film system was designed to modulate the releasing rate of the drug. The top film equilibrated with 0.75 or 1.5 M NaCl release up to 41.9% and 89.0% of the enrofloxacin in 4 h, respectively. The release rate of enrofloxacin was found dependent on NaCl concentration in the upper gel layer. The two-layer cryogel system showed attractive features for transcutaneous antibiotic delivery.

Transglycosylation specificity of Acremonium sp. α-rhamnosyl-ß-glucosidase and its application to the synthesis of the new fluorogenic substrate 4-methylumbelliferyl-rutinoside.

Transglycosylation potential of the fungal diglycosidase α-rhamnosyl-ß-glucosidase was explored. The biocatalyst was shown to have broad acceptor specificity toward aliphatic and aromatic alcohols. This feature allowed the synthesis of the diglycoconjugated fluorogenic substrate 4-methylumbelliferyl-rutinoside. The synthesis was performed in one step from the corresponding aglycone, 4-methylumbelliferone, and hesperidin as rutinose donor. 4-Methylumbelliferyl-rutinoside was produced in an agitated reactor using the immobilized biocatalyst with a 16% yield regarding the sugar acceptor. The compound was purified by solvent extraction and silica gel chromatography. MALDI-TOF/TOF data recorded for the [M+Na](+) ions correlated with the theoretical monoisotopic mass (calcd [M+Na](+): 507.44 m/z; obs. [M+Na](+): 507.465 m/z). 4-Methylumbelliferyl-rutinoside differs from 4-methylumbelliferyl-glucoside in the rhamnosyl substitution at the C-6 of glucose, and this property brings about the possibility to explore in nature the occurrence of endo-ß-glucosidases by zymographic analysis.

Extracellular monoenzyme deglycosylation system of 7-O-linked flavonoid beta-rutinosides and its disaccharide transglycosylation activity from Stilbella fimetaria.

We screened for microorganisms able to use flavonoids as a carbon source; and one isolate, nominated Stilbella fimetaria SES201, was found to possess a disaccharide-specific hydrolase. It was a cell-bound ectoenzyme that was released to the medium during conidiogenesis. The enzyme was shown to cleave the flavonoid hesperidin (hesperetin 7-O-alpha-rhamnopyranosyl-beta-glucopyranoside) into rutinose (alpha-rhamnopyranosyl-beta-glucopyranose) and hesperetin. Since only intracellular traces of monoglycosidase activities (beta-glucosidase, alpha-rhamnosidase) were produced, the disaccharidase alpha-rhamnosyl-beta-glucosidase was the main system utilized by the microorganism for hesperidin hydrolysis. The enzyme was a glycoprotein with a molecular weight of 42224 Da and isoelectric point of 5.7. Even when maximum activity was found at 70 degrees C, it was active at temperatures as low as 5 degrees C, consistent with the psychrotolerant character of S. fimetaria. Substrate preference studies indicated that the enzyme exhibits high specificity toward 7-O-linked flavonoid beta-rutinosides. It did not act on flavonoid 3-O-beta-rutinoside and 7-O-beta-neohesperidosides, neither monoglycosylated substrates. In an aqueous medium, the alpha-rhamnosyl-beta-glucosidase was also able to transfer rutinose to other acceptors besides water, indicating its potential as biocatalyst for organic synthesis. The monoenzyme strategy of Acremonium sp. SES201 = DSM 24697, [corrected] as well as the enzyme substrate preference for 7-O-beta-flavonoid rutinosides, is unique characteristics among the microbial flavonoid deglycosylation systems reported.