Use of Novel Homochiral Thioureas Camphor Derived as Asymmetric Organocatalysts in the Stereoselective Formation of Glycosidic Bonds.

We synthesized six new camphor-derived homochiral thioureas 1-6, from commercially available (1R)-(-)-camphorquinone. These new compounds 1-6 were evaluated as asymmetric organocatalysts in the stereoselective formation of glycosidic bonds, with 2,3,4,6-tetra-O-benzyl-D-glucopyranosyl and 2,3,4,6-tetra-O-benzyl-D-galactopyranosyl trichloroacetimidates as donors, and several alcohols as glycosyl acceptors, such as methanol, ethanol, 1-propanol, 1-butanol, 1-octanol, iso-propanol, tert-butanol, cyclohexanol, phenol, 1-naphtol, and 2-naphtol. Optimization of the asymmetric glycosylation reaction was achieved by modifying reaction conditions such as solvent, additive, loading of catalyst, temperature, and time of reaction. The best result was obtained with 2,3,4,6-tetra-O-benzyl-D-galactopyranosyl trichloroacetimidates, using 15 mol% of organocatalyst 1, in the presence of 2 equiv of MeOH in solvent-free conditions at room temperature for 1.5 h, affording the glycosidic compound in a 99% yield and 1:73 α:ß stereoselectivity; under the same reaction conditions, without using a catalyst, the obtained stereoselectivity was 1:35 α:ß. Computational calculations prior to the formation of the products were modeled, using density functional theory, M06-2X/6-31G(d,p) and M06-2X/6-311++G(2d,2p) methods. We observed that the preference for ß glycoside formation, through a stereoselective inverted substitution, relies on steric effects and the formation of hydrogen bonds between thiourea 1 and methanol in the complex formed.

Stabilization of Anthocyanins from Coffee (Coffea arabica L.) Husks and In Vivo Evaluation of Their Antioxidant Activity.

Coffee (Coffea arabica L.) is one of the most popular and widely consumed products throughout the world, mainly due to its taste, aroma, caffeine content, and natural antioxidants. Among those antioxidants, anthocyanins are one of the most important natural pigments, which can be found in coffee husks. It is widely known that anthocyanins have multiple health benefits partially linked to their antioxidant properties. However, anthocyanins have low stability and are sensitive to all types of changes. In order to prevent its degradation, anthocyanins can be stabilized with nanoparticles. Thus, the main objective of this study was to evaluate the stability of the anthocyanins extracted from coffee husks, using three different extracting agents (ethanol, methanol, and water) and stabilizing them through conjugation with zinc oxide nanoparticles. The anthocyanins extracts were mainly composed of cyanidin-3-rutinoside (97%) and the total phenolic compounds of the fresh extracts were 458.97 ± 11.32 (methanol), 373.53 ± 12.74 (ethanol), and 369.85 ± 15.93 (water) mg GAE/g. On the other hand, the total phenolic compounds of the nanoparticle-anthocyanin conjugates underwent no significant changes after stabilization as the major loss was less than 3%. Furthermore, the percentage of anthocyanins' degradation was less than 5% after 12 weeks of storage. On top of that, fresh anthocyanin extracts and anthocyanin-nanoparticle conjugates exhibited a strong protective effect against oxidative stress and increased the survival rate of Caenorhabditis elegans.

A dopamine-based biosynthetic pathway produces decarboxylated betalains in Chenopodium quinoa.

Betalains are the nitrogenous pigments that replace anthocyanins in the plant order Caryophyllales. Here, we describe unconventional decarboxylated betalains in quinoa (Chenopodium quinoa) grains. Decarboxylated betalains are derived from a previously unconsidered activity of the 4,5-DOPA-extradiol-dioxygenase enzyme (DODA), which has been identified as the key enzymatic step in the established biosynthetic pathway of betalains. Here, dopamine is fully characterized as an alternative substrate of the DODA enzyme able to yield an intermediate and structural unit of plant pigments: 6-decarboxy-betalamic acid, which is proposed and described. To characterize this activity, quinoa grains of different colors were analyzed in depth by chromatography, time-of-flight mass spectrometry, and reactions were performed in enzymatic assays and bioreactors. The enzymatic-chemical scheme proposed leads to an uncharacterized family of 6-decarboxylated betalains produced by a hitherto unknown enzymatic activity. All intermediate compounds as well as the final products of the dopamine-based biosynthetic pathway of pigments have been unambiguously determined and the reactions have been characterized from the enzymatic and functional perspectives. Results evidence a palette of molecules in quinoa grains of physiological relevance and which explain minor betalains described in plants of the Caryophyllales order. An entire family of betalains is anticipated.

Nuclear Magnetic Resonance Metabolomics with Double Pulsed-Field-Gradient Echo and Automatized Solvent Suppression Spectroscopy for Multivariate Data Matrix Applied in Novel Wine and Juice Discriminant Analysis.

The quality of foods has led researchers to use various analytical methods to determine the amounts of principal food constituents; some of them are the NMR techniques with a multivariate statistical analysis (NMR-MSA). The present work introduces a set of NMR-MSA novelties. First, the use of a double pulsed-field-gradient echo (DPFGE) experiment with a refocusing band-selective uniform response pure-phase selective pulse for the selective excitation of a 5-10-ppm range of wine samples reveals novel broad 1H resonances. Second, an NMR-MSA foodomics approach to discriminate between wine samples produced from the same Cabernet Sauvignon variety fermented with different yeast strains proposed for large-scale alcohol reductions. Third a comparative study between a nonsupervised Principal Component Analysis (PCA), supervised standard partial (PLS-DA), and sparse (sPLS-DA) least squares discriminant analysis, as well as orthogonal projections to a latent structures discriminant analysis (OPLS-DA), for obtaining holistic fingerprints. The MSA discriminated between different Cabernet Sauvignon fermentation schemes and juice varieties (apple, apricot, and orange) or juice authentications (puree, nectar, concentrated, and commercial juice fruit drinks). The new pulse sequence DPFGE demonstrated an enhanced sensitivity in the aromatic zone of wine samples, allowing a better application of different unsupervised and supervised multivariate statistical analysis approaches.

Chemistry, Occurrence, Properties, Applications, and Encapsulation of Carotenoids-A Review.

Carotenoids are natural lipophilic pigments and antioxidants that are present in many fruits and vegetables. The consumption of carotenoids is correlated with positive health effects and a decreased risk of several chronic diseases. Provitamin A carotenoids (ß-carotene, α-carotene, γ-carotene, and ß-cryptoxanthin) are essential for the development and maintenance of sight. ß-carotene, α-carotene, zeaxanthin, ß-cryptoxanthin, lutein, and lycopene have high antioxidant activity and promote free radical scavenging, which helps protect against chronic diseases. However, carotenoids are chemically unstable and prone to oxidation in the presence of light, heat, oxygen, acids, and metal ions. The use of carotenoids in the food industry is limited due to their poor solubility in water, bioavailability and quick release. Encapsulation techniques, such as microencapsulation, nanoencapsulation and supercritical encapsulation, are used to overcome these problems. The objective of this paper is to describe the characteristics and potential health benefits of carotenoids and advances in encapsulation techniques for protecting and enhancing their solubility or bioavailability.

Carotenoids from mamey (Pouteria sapota) and carrot (Daucus carota) increase the oxidative stress resistance of Caenorhabditis elegans.

Carotenoids are natural pigments and antioxidants found in fruits and vegetables such as carrot, tomato, orange, mango, yellow corn, pumpkin, and mamey. In this study, we evaluated the antioxidant potential of mamey (Pouteria sapota) carotenoids and compared them to carrot (Daucus carota) carotenoids. The carotenoids were extracted from mamey and carrot, and their antioxidant capacity were determined via in vitro (ABTS method) and in vivo assays (resistance against oxidative stress in Caenorhabditis elegans). The carotenoid contents in mamey and carrot were 4.42 ± 0.12 and 5.47 ± 0.04 mg ß-carotene/100 g, respectively. Despite the differences between the carotenoid contents in both products (p < 0.05), the in vitro antioxidant capacity results showed no significant differences between the extracts (p > 0.05). The mamey and carrot carotenoid extracts decreased the oxidative damage in C. elegans by 20-30% and 30-40%, respectively. Both extracts increased the resistance and enhanced the survival of the nematodes, and showed better effects than pure ß-carotene, probably owing to the complex mixture in the carotenoid extracts. These results suggest that mamey is a good alternative source of carotenoids and that it protects against oxidative stress in C. elegans. The protective effect of mamey carotenoids was similar to the effect of carrot carotenoids.

First Betalain-Producing Bacteria Break the Exclusive Presence of the Pigments in the Plant Kingdom.

The biosynthesis of antioxidant pigments, namely, betalains, was believed to be restricted to Caryophyllales plants. This paper changes this paradigm, and enzyme mining from bacterial hosts promoted the discovery of bacterial cultures producing betalains. The spectrum of possible sources of betalain pigments in nature is broadened by our description of the first betalain-forming bacterium, Gluconacetobacter diazotrophicus The enzyme-specific step is the extradiol cleavage of the precursor amino acid l-dihydroxyphenylalanine (l-DOPA) to form the structural unit betalamic acid. Molecular and functional work conducted led to the characterization of a novel dioxygenase, a polypeptide of 17.8 kDa with a Km of 1.36 mM, with higher activity and affinity than those of its plant counterparts. Its superior activity allowed the first experimental characterization of the early steps in the biosynthesis of betalains by fully characterizing the presence and time evolution of 2,3- and 4,5-seco-DOPA intermediates. Furthermore, spontaneous chemical reactions are characterized and incorporated into a comprehensive enzymatic-chemical mechanism that yields the final pigments.IMPORTANCE Several studies have demonstrated the health-promoting effects of betalains due to their high antioxidant capacity and their positive effect on the dose-dependent inhibition of cancer cells and their proliferation. To date, betalains were restricted to plants of the order Caryophyllales and some species of fungi, but the present study reveals the first betalain-producing bacterium, as well as the first steps in the formation of pigments. This finding demonstrates that betalain biosynthesis can be expanded to prokaryotes.

A colorimetric assay for the determination of acetyl xylan esterase or cephalosporin C acetyl esterase activities using 7-amino cephalosporanic acid, cephalosporin C, or acetylated xylan as substrate.

A bromothymol blue-based colorimetric assay has been devised to screen for acetyl xylan esterase or cephalosporin C (CPC) deacetylase activities using 7-amino cephalosporanic acid (7-ACA), CPC, or acetylated xylan as substrate. These enzymes are not screened with their natural substrates because of the tedious procedures available previously. Acetyl xylan esterase from Bacillus pumilus CECT 5072 was cloned, expressed in Escherichia coli Rosetta (DE3), and characterized using this assay. Similar K(M) values for 7-ACA and CPC were obtained when compared with those described using HPLC methods. The assay is easy to perform and can be carried out in robotic high-throughput colorimetric devices normally used in directed evolution experiments. The assay allowed us to detect improvements in activity at a minimum of twofold with a very low coefficient of variance in 96-well plates. This method is significantly faster and more convenient to use than are known HPLC and pH-stat procedures.