Occurrence and sensory perception of Z-2-(β-d-glucopyranosyloxy)-3-phenylpropenoic acid in rooibos (Aspalathus linearis)

Z-2-(β-d-glucopyranosyloxy)-3-phenylpropenoic acid (PPAG), a compound postulated to contribute to the taste and mouthfeel of fermented rooibos tea (Aspalathus linearis), was isolated from unfermented rooibos plant material. Its structure was unequivocally confirmed by LC–MS, –MS², FT-IR and NMR of the underivatised natural product, and optical rotation measurements of the hydrolysed sugar moiety. A similar compound, postulated to be E-2-(β-d-glucopyranosyloxy)-3-phenylpropenoic acid, was also detected. Analysis of the leaves of a large number of rooibos plants (n = 54), sampled at commercial plantations, showed that PPAG is not ubiquitously present in detectable quantities in the leaves of different plants. This leads to large variation in the fermented plant material, infusions and food-grade extracts. PPAG was shown to have a slightly bitter to astringent taste and a detection threshold of 0.4 mg/l in water.     

Sugars,acids, ethyl β-d-glucopyranose and a methyl inositol in sea buckthorn (Hippophaë rhamnoides) berries

Sea buckthorn berry is a rich source of nutrients and bioactive components beneficial for human health. Sugars and acids play an important role in determining the sensory properties of the berry. Sugars, acids, ethyl β-d-glucopyranose and a methyl inositol were analysed in berries of three subspecies (Hippophaë rhamnoides ssp. sinensis, rhamnoides and mongolica) collected from China, Finland and Russia over four consecutive years. Fructose and glucose were the major sugars, and the dominating acids were malic and quinic acids. Origin and harvesting date have significant impacts on sugars, acids and sugar/acid ratio in the berry. During the harvesting period, the sugar content followed different changing patterns in berries of different subspecies. Ethyl glucose dominated in the sugar fraction of ssp. rhamnoides but existed only in trace amounts in the other two subspecies. In ssp. rhamnoides, the level of ethyl glucose increased during the harvesting period; the increase was accompanied by a decrease in glucose content, indicating the presence of a biochemical pathway converting glucose into its derivatives. A methyl inositol was identified for the first time in sea buckthorn with higher levels found in ssp. sinensis than in the other two subspecies. The levels of ethyl glucose and methyl inositol may be important sensory and nutritional quality factors of sea buckthorn berry. The data presented by this study provide important chemotaxonomic information characterising different subspecies of sea buckthorn and useful guidance for breeding, harvesting, and industrial utilisation of sea buckthorn.     

Determination of l- and d-carnitine in dietary food supplements using capillary electrophoresis–tandem mass spectrometry

This paper reports the application of a capillary electrophoresis–electrospray ionisation-tandem mass spectrometry methodology for the unequivocal identification and the quantitative determination of the two enantiomers of the non-protein amino acid carnitine (l- and d-Carn) in 22 dietary food supplements, including drinks, biscuits, capsules and tablets. MS/MS experiments were optimised to achieve the high sensitivity and selectivity required for food analysis. A comparison of the slopes obtained by the external standard and the standard additions calibration methods indicated the absence of matrix interferences in these food samples. Good precision (RSDs ranged from 2.3% to 4.3% for migration times, and from 2.1% to 10.5% for corrected peak areas) and acceptable accuracy established by means of recovery studies (from 85% to 102%) were obtained. The limit of detection was about 10 ng/mL (S/N = 3) enabling the determination of enantiomeric impurities (d-Carn) up to 0.025% of Carn in foods. This chiral method illustrated is suitable for routine qualitative and quantitative analyses of Carn in foods. The results showed contents for l-Carn comprised from 47% to 115% with respect to the labelled content of l-Carn. Percentages obtained for the d-enantiomer ranged from 0.4% to 5.9%, except for one of the samples analysed, that contained the racemate (49.3% d-Carn). The use of the racemate is not allowed by legislation, which corroborated the high potential of the developed method in the control of the quality and safety of foods containing Carn.     

Comparison of the solution properties of (1→3),(1→4)-β-d-glucans extracted from oats and barley

The solution properties of β-glucan isolated from whole grains of oats and barley were studied by viscosity and oscillatory measurements both in water and in aqueous cuoxam, an ammoniacal Cu(OH)₂/CuCl-solution. The viscosity measurements of water solutions show stronger shear thinning for oat than for barley β-glucans. The crossover points of the moduli G′ and G″ in cuoxam solutions were different for the two β-glucans, i.e. 10 Hz for oat β-glucan and 1.5 Hz for barley β-glucan. The molecular weights of the β-glucans were similar. Therefore, the difference in solution properties seems to be related to the difference in structure. This difference is revealed by the ratio DP3:DP4 of the main building blocks of β-glucans, which was higher for barley than for oat β-glucan.     

Production of d-galactose using β-galactosidase and Saccharomyces cerevisiae entrapped in poly(vinylalcohol) hydrogel

d-Galactose was produced from lactose (200 g l⁻¹) in the batch mode of a simultaneous saccharification and fermentation process (SSF). β-Galactosidases (from Kluyveromyces lactis and Aspergillus oryzae) and yeasts were immobilized in poly(vinylalcohol) hydrogel lens-shaped capsules – LentiKats^®. After 20 repeated batch runs with entrapped K. lactis, β-galactosidase and free Saccharomyces cerevisiae (10% v/v inoculum), galactose productivity decreased to 50% and 1.4 kg of galactose were prepared. Compared to this, just 20% decrease of galactose productivity and a 0.9 kg production of galactose were observed for the SSF process with β-galactosidase from A. oryzae after 15 repeated batches under the same conditions. In the process of SSF with co-immobilized enzyme from K.lactis and S.cerevisiae, the galactose productivity increased from 3 g l⁻¹ h⁻¹ to 4.1 g l⁻¹ h⁻¹, which reduced the time of preparation of d-galactose.     

Antioxidative activity of (1→3), (1→6)-β-d-glucan from Saccharomyces cerevisiae grown on different media

In many microorganisms, fungi and algae (1→3), (1→6)-β-d-glucan can be found as cell wall polysaccharide. Various publications describe the strong positive influence of these glucans on the immune system comprising antibacterial, wound-healing and antitumour activities. Recently, it was found that (1→3)-β-d-glucan from the yeast Saccharomyces cerevisiae also exhibits antioxidative capabilities.In the present study, the antioxidative activity of glucan from the cell walls of S. cerevisiae grown on different media (wort, yeast-peptone-glucose, yeast-peptone-galactose) was investigated.Fermentation of the yeast took place on three different media. After fermentation the yeast was harvested and disrupted in a high-pressure homogeniser. Subsequently, glucan was extracted from the yeast cell walls by a procedure including a combination of hot water and enzymatic treatment. The level of (1→3), (1→6)-β-d-glucan in the cell walls was analysed enzymatically. The antioxidant activity was determined by electron paramagnetic resonance (EPR) spectrometry, TEAC assay (Trolox equivalent antioxidant capacity) and an ‘amperometric biosensor’.The results show significant differences in the glucan content of the cell walls and in their antioxidative activities depending on growth medium. However, glucan itself seems to have a low antioxidative activity in contrast to other cell wall fractions e.g. proteins.     

α-l-Arabinofuranosidase 3 from Penicillium purpurogenum (ABF3): Potential application in the enhancement of wine flavour and heterologous expression of the enzyme

An α-l-arabinofuranosidase (ABF3) from Penicillium purpurogenum was purified and its possible biotechnological application in the enhancement of wine flavour combined with P. purpurogenum β-glucosidase was studied. A must from Muscat of Alexandria was used to isolate the glycosides. The total monosaccharide (glucose, arabinose and xylose) levels in the glycosides were determined after acid hydrolysis, and were compared with the result of enzymatic hydrolysis. These results were analogous to those obtained in similar experiments using a commercial preparation, thus suggesting that the enzyme from P. purpurogenum may prove useful in this particular application. This prompted us to express the enzyme heterologously. The abf3 gene was thus expressed in Pichia pastoris. The recombinant enzyme was purified and it shows the same properties of the native ABF3 (substrate specificity, kinetic constants, pH and temperature optima and antibody cross-reactivity).     

The effects of MgSO4, d-glucono-δ-lactone (GDL), sucrose,and urea on gelation properties of pectic polysaccharide from soy hull

The monosaccharide composition of pectic polysaccharide extracted from soy hull under assistance of microwave was firstly analyzed. Soy hull pectic polysaccharide (SHPP) mainly contains galactose, xylose, galacturonic acid, arabinose, glucose and rhamnose with a molar ratio of 4.7:2.5:1.5:3.8:3.2:3.0. The gelation properties of SHPP as a function of Mg²⁺, GDL, sucrose, and urea concentration were then systematically studied. Mg²⁺ can induce the gelation of SHPP but low concentration Mg²⁺ require more time to cross-linking SHPP molecular chains. GDL can strengthen the elasticity of SHPP/Mg²⁺ gel, while sucrose can also strengthen the network through hydrophobic interaction, however 30 wt% sucrose can weaken the SHPP/Mg²⁺ gel network. 2 mol/L urea can collapse the gel network formed by 3 wt% GDL with 0.1 mol/L Mg²⁺, which suggests that the junction zone of SHPP/GDL/Mg²⁺ system is formed through hydrogen bonding.     

Effects of d-α-tocopherol and dietary energy on growth and health of preruminant dairy calves

To observe the effects of supplemental dietary d-α-tocopherol in relation to dietary energy on growth and immune status in dairy calves, 32 newborn Holstein bull calves were assigned to 1 of 4 treatments for 5 wk in a 2 × 2 factorial, randomized complete block, split-plot design. Calves received moderate growth (MG) or low growth (LG) all-milk dietary treatments, formulated to support daily gains of 0.5 or 0.25 kg/d, respectively, per the dietary energy recommendation for milk-fed calves according to the National Research Council’s Nutrient Requirements of Dairy Cattle. Calves in both groups were either injected i.m. with Vital E-A+D (injectable solution of vitamins E, A, and D) on d 1 and supplemented with Emcelle Tocopherol (micellized vitamin E) via milk daily (MG-S and LG-S), or were not supplemented (MG-C and LG-C) during the study period. Total weight gain of MG calves was greater than that of LG calves and tended to be greater in MG-S calves than in MG-C calves. Calves receiving vitamin supplementation demonstrated greater concentrations of plasma α-tocopherol, retinol, and 25-(OH)-vitamin D than did control calves, whereas MG calves demonstrated a lower concentration of plasma α-tocopherol than did LG calves. The apparent increased utilization of α-tocopherol by MG calves was accompanied by a rise in serum haptoglobin, a positive acute-phase protein and indicator of inflammation, especially in MG-C calves. Serum amyloid A, also a positive acute-phase protein, was not different among groups, but was elevated from baseline in all groups during wk 1 through 3. Plasma IgG₁ concentrations were higher in MG-S and LG-S calves than in their nonsupplemented dietary counterparts, whereas plasma IgG₂, IgA, and IgM concentrations were not different among groups. In summary, dietary supplementation of d-α-tocopherol improved plasma α-tocopherol status and tended to increase growth in calves fed for 0.5 kg of average daily gain. Vitamin supplementation ameliorated the rise of serum haptoglobin associated with acute inflammation in MG calves, and may have improved passive transfer of maternal antibody. These results indicate a role for α-tocopherol in prevention of proinflammatory state associated with greater dietary energy and onset of infectious disease.     

Antioxidant and antiacetylcholinesterase activities of essential oils from Cymbopogon schoenanthus L. Spreng. Determination of chemical composition by GC–mass spectrometry and C NMR

Cymbopogon schoenanthus L. Spreng, is an aromatic herb consumed in salads and used to prepare traditional meat recipes in Tunisia. The chemical composition, antioxidant activities and acetylcholinesterase inhibitory properties of the essential oils from fresh leaves, dried leaves and roots collected from three different locations in southern Tunisia, were evaluated. Essential oils were analysed by GC–mass spectrometry and ¹³C NMR. The major components were limonene (10.5–27.3%), β-phellandrene (8.2–16.3%), δ-terpinene (4.3–21.2%) and α-terpineol (6.8–11.0%). Antioxidant activity was measured by DPPH assay. The results ranged from 36.0% to 73.8% (2 μl of essential oil per mL of test solution).     

Aggregation and structural changes of silver carp actomyosin as affected by mild acidification with d-gluconic acid δ-lactone

The structural changes and aggregation properties of silver carp actomyosin acidified with d-gluconic acid-δ-lactone (GDL) were investigated. Results showed that silver carp actomyosin underwent aggregation and formation of precipitate as indicated by turbidity and centrifugation coupled electrophoresis analysis. Circular dichroism indicated that myosin rod unfolded during acidification, resulting in a gradual decrease in α-helical content. The changes in tertiary structure of actomyosin under acidic conditions were demonstrated by second-derivative UV spectra and intrinsic fluorescence. Tyrosine residues were exposed to the surface of proteins when pH was decreased to 5.5, and were buried inside the protein aggregates with further reduction in pH. In contrast, more tryptophan residues were exposed to the polar environment with decreasing pH. 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide crosslinking experiments showed that the intensity of myosin heavy chain (MHC) bands decreased sharply with decreasing pH and the actin bands decreased more slowly, suggesting that MHC is the major protein component involved in the non-covalent cross-linking and formation of aggregates during acidification of silver carp actomyosin.     

Short communication: Nα-Lauroyl-l-arginine ethylester monohydrochloride reduces bacterial growth in pasteurized milk

Effective strategies for extending fluid milk product shelf-life by controlling bacterial growth are of economic interest to the dairy industry. To that end, the effects of addition of l-arginine, Nα-lauroyl ethylester monohydrochloride (LAE) on bacterial numbers in fluid milk products were measured. Specifically, LAE was added (125, 170, or 200 mg/L) to conventionally homogenized and pasteurized 3.25% fat chocolate or unflavored milk products. The treated milks and corresponding untreated controls were held at 6°C and plated on standard plate count agar within 24 h of processing and again at 7, 14, 17, and 21 d of storage. Bacterial counts in all unflavored milk samples treated with LAE remained below the Pasteurized Milk Ordinance limit for grade A pasteurized fluid milk of 4.3 log cfu/mL for the entire 21 d. Bacterial counts in unflavored samples containing 170 and 200 mg/L of LAE were significantly lower than those in the untreated unflavored milk at d 17 and 21 postprocessing. Specifically, bacterial counts in the milk treated with 200 mg/L of LAE were 5.77 log cfu/mL lower than in untreated milk at 21 d postprocessing. Bacterial counts in chocolate milk treated with 200 mg/L of LAE were significantly lower than those in the untreated chocolate milk at d 14, 17, and 21. In chocolate milk treated with 200 mg/L of LAE, bacterial counts were 0.9 log cfu/mL lower than in the untreated milk at 21 d postprocessing. Our results show that addition of LAE to milk can reduce bacterial growth. Addition of LAE is more effective at controlling bacterial growth in unflavored milk than in chocolate milk.     

Migration of α-tocopherol and resveratrol from poly(L-lactic acid)/starch blends films into ethanol

Poly(L-lactic acid) (PLLA)/starch blends with various concentrations of two natural antioxidants, α-tocopherol (α-TOC) and resveratrol, were fabricated by a melt blending and compression molding processes. The effects of the two antioxidants on the optical (color), thermal and mechanical properties of PLLA/starch blends with antioxidants were assessed. PLLA/starch blend films with α-TOC and resveratrol showed a yellowish color influenced by the combined effect of white starch and the brown color of the antioxidants. The glass transition and melting temperatures were significantly reduced with the addition of antioxidants while enhanced thermal stability was observed, which could be a benefit and important for processing and production. The enhanced mechanical properties could be attributed to not only a compatibilization effect based on the chemical linkage between PLLA and starch chains, but also restriction of the chain mobility by antioxidants. The release of resveratrol from PLLA and PLLA/starch blend films into ethanol followed Fickian behavior. The D values of α-TOC were in the range of 0.47–3.95 × 10⁻¹¹ cm² s⁻¹ for PLLA films and 0.70–6.83 × 10⁻¹¹ cm² s⁻¹ for PLLA/starch blend films at 13 °C, 5.67–13.0 × 10⁻¹¹ cm² s⁻¹ for PLLA films and 4.10–24.2 × 10⁻¹¹ cm² s⁻¹ for PLLA/starch blend films at 23 °C, and 89.0–118.0 × 10⁻¹¹ cm² s⁻¹ for PLLA films and 123–282 × 10⁻¹¹ cm² s⁻¹ for PLLA/starch blend films at 43 °C. The D values of resveratrol were in the range of 0.073–0.54 × 10⁻¹⁰ cm² s⁻¹ for PLLA films and 1.42–6.93 × 10⁻¹⁰ cm² s⁻¹ for PLLA/starch blend films at 13 °C, 0.90–3.44 × 10⁻¹⁰ cm² s⁻¹ for PLLA films and 4.16–22.3 × 10⁻¹⁰ cm² s⁻¹ for PLLA/starch blend films at 23 °C, and 24.8–74.1 × 10⁻¹⁰ cm² s⁻¹ for PLLA films and 40.1–309 × 10⁻¹⁰ cm² s⁻¹ for PLLA/starch blend films at 43 °C.     

Hydrolysis of soybean isoflavone glycosides by a thermostable β-glucosidase from Paecilomycesthermophila

The β-glucosidase from Paecilomyces thermophila J18 was found to be capable of hydrolysing daidzin and genistin in a previous study. This report further evaluated the thermostability and hydrolysis of soybean isoflavone glycosides. The enzyme was found to be very stable at 50 °C, and retained more than 95% of its initial activity after 8 h at 50 °C. It converted isoflavone glycosides, in soybean flour extract and soybean embryo extract, to their aglycones, resulting in more than 93% of hydrolysis of three isoflavone glycosides (namely, daidzin, genistin and glycitin) after 4 h of incubation. Also, addition of the β-glucosidase greatly increased the contents of isoflavone aglycones in the suspended soybean flour and soymilk. The results indicate that the thermostable β-glucosidase may be used to increase the isoflavone aglycones in soy products. This is the first report on the potential application of fungal β-glucosidases for converting isoflavone glycosides to their aglycones in soy products.     

The contents of the neuro-excitatory amino acid β-ODAP (β-N-oxalyl-l-α,β-diaminopropionic acid), and other free and protein amino acids in the seeds of different genotypes of grass pea (Lathyrus sativus L.)

The free and protein amino acids of nine different genotypes of grass pea (Lathyrus sativus L.) seeds were analysed by HPLC with pre-column PITC (phenyl isothiocyanate) derivatisation. Among the free amino acids, homoarginine was quantitatively the most important (up to 0.8% seed weight) and stable while the neuro-excitatory amino acid β-ODAP (β-N-oxalyl-l-α,β-diaminopropionic acid) showed highest variation (0.02–0.54%) in the nine genotypes examined. Among protein amino acids, glutamic acid was quantitatively most significant, followed by aspartic acid, arginine, leucine, lysine and proline. The sulphur amino acid, methionine, showed the lowest concentration in all the L. sativus genotypes, and also in lentil (Lens culinaris) and in soybean (Glycine max) seeds analysed at the same time.     

Characterization of phenolic compounds in Chinese hawthorn (Crataegus pinnatifida Bge. var. major) fruit by high performance liquid chromatography–electrospray ionization mass spectrometry

Hawthorn (Crataegus spp.) fruits are increasingly popular as raw materials for nutraceuticals and functional foods. As a major group of bio-active components of hawthorn, the phenolic compounds of the fruits have not been well characterized so far. After extraction with 80% aqueous ethanol, the phenolics of the fruits of a major Chinese hawthorn variety, Crataegus pinnatifida Bge. var. major, were separated by polyamide column chromatography, followed by analyses by high performance liquid chromatography combined with diode array UV spectrometry and electrospray ionization mass spectrometry. Ideain (cyanidin-3-O-galactoside), chlorogenic acid, procyanidin B2 [epicatechin-(4β → 8)-epicatechin], epicatechin, hyperoside (quercetin-3-O-galactoside) and isoquercitrin (quercetin-3-O-glucoside) were identified with UV spectra, mass spectra and reference compounds. In addition, 35 compounds were tentatively identified based on UV and mass spectra. These compounds were mostly B-type procyanidins (PA) and their glycosides including aglycons of 3 dimers, 3 trimers, 8 tetramers, 4 pentamers, 2 hexamers and 2 glycosides of PA monomers, 7 glycosides of PA dimers, 1 glycoside of a PA trimer, 2 glycosides of PA tetramers, 1 glycoside of a PA pentamer, and 2 glycosides of quercetin. This is the first systematic study of phenolic compounds in Chinese hawthorn fruits and the first report of the presence of glycosylated procyanidins in hawthorn.