Free Sinapic Acid Research Articles

Grafting of sinapic acid onto glucosamine nanoparticle as a potential therapeutic drug with enhanced anti-inflammatory activities in osteoarthritis treatment

Glucosamine (Glu) is a cartilage and joint fluid matrix precursor that modulates osteoarthritic joint changes. To improve the enzymatic stability, glucosamine was developed into nanoglucosamine by the ionic gelation method through sodium tripolyphosphate (TPP) as cross-linking agent. The optimized mass ratio of Glu:TPP was (3:1) with the particle size 163 ± 25 nm and surface charge −5 mV. Then Sinapic acid (SA) as a natural phenolic acid with strong antioxidant and antimicrobial activities has been grafted onto glucosamine nanoparticles (GluNPs) with grafting efficiency (73 ± 6 %). The covalent insertion of SA was confirmed by UV–Vis, FTIR, 1HNMR, XRD, and FESEM analyses and the other physicochemical properties were also characterized. SA-g-GluNPs showed spherical shape with a mean diameter of 255 ± 20 nm and zeta potential +16 mV. The in vitro release profile of SA-g-GluNPs exhibited the sustained and pH-dependent drug release property. SA-g-GluNPs had a more pronounced effect on reducing the elevated levels of LPS-induced oxidative stress and pro-inflammatory cytokines than free SA in the human chondrocyte C28/I2 cell line. Furthermore, the antibacterial properties against E. coli and S. aureus were also improved by SA-g-GluNPs. This study demonstrated the potential of phenolic acid grafted GluNPs in therapeutic drug applications for chondroprotection and food industries.

Cell dehydration of intergeneric hybrid induces subgenome-related specific responses.

The aim was to identify subgenome-related specific responses in two types of triticale, that is, of the wheat-dominated genome (WDG) and rye-dominated genome (RDG), to water stress induced in the early phase (tillering) of plant growth. Higher activity of the primary metabolism of carbohydrates is a feature of the WDG type, while the dominance of the rye genome is associated with a higher activity of the secondary metabolism of phenolic compounds in the RDG type. The study analyzed carbohydrates and key enzymes of their synthesis, free phenolic compounds and carbohydrate-related components of the cell wall, monolignols, and shikimic acid (ShA), which is a key link between the primary and secondary metabolism of phenolic compounds. Under water stress, dominance of the wheat genome in the WDG type was manifested by an increased accumulation of the large subunit of Rubisco and sucrose phosphate synthase and a higher content of raffinose and stachyose compared with the RDG type. In dehydrated RDG plants, higher activity of L-phenylalanine ammonia lyase (PAL) and L-tyrosine ammonia lyase (TAL), as well as a higher level of ShA, free and cell wall-bound p-hydroxybenzoic acid, free homovanillic acid, free sinapic acid, and cell wall-bound syringic acid can be considered biochemical indicators of the dominance of the rye genome.

Preparation, development and characterization of Leucaena leucocephala galactomannan (LLG) conjugated sinapic acid: A potential colon targeted prodrug

Sinapic acid (SA), a widely prevalent hydroxycinnamic acid, possess numerous biological activities owing to its antioxidant property. The present study was aimed to prepare colon targeted polysaccharidic/polymeric ester prodrug of SA (a microbially triggered system) using Leucaena leucocephala galactomannan (LLG) as a polysaccharidic carrier. The polymeric conjugates of SA-LLG were found to exhibit an increase in % yield and DS with increase in amount of SA and volume of thionyl chloride. The degree of depolymerization of SA-LLG prodrug batches were evaluated using optimized concentration of galactomannase. The SA-LLG prodrug was characterized employing UV and FTIR spectroscopy, 1H NMR and XRD. In vitro release study of the optimized prodrug batch (SL10) suggested stable nature of SA-LLG conjugate under acidic (pH 1.2) and alkaline conditions (pH 6.8). The treatment of prodrug with galactomannase (15 mg/mL) followed by esterase (10 U/mL) enzyme released approximately 81% of SA after 24 h. The cell viability results revealed that free SA and SA-LLG were found to have similar antiproliferative potential against human colon cancer cell lines (HCT-116 cells). Our investigation revealed that polysaccharidic prodrug, SA-LLG, has the potential for colon targeting of SA and thus can be employed for the treatment of Inflammatory Bowel Diseases (IBDs).

Release of phenolic acids from sunflower and rapeseed meals using different carboxylic esters hydrolases from Aspergillus niger

Sunflower and rapeseed meals are agro-industrial coproducts that contain high amount of phenolics (1–4 % dry matter), mostly as esters of caffeic acid (CA) and sinapic acid (SA), respectively. The enzymatic hydrolysis of the ester bonds enables to recover the corresponding free phenolic acids that are bioactive compounds and platform molecules for various applications in green chemistry. Here we aimed to find the best route for producing free CA and SA by applying various fungal carboxylic ester hydrolases from recombinant Aspergillus niger strains either directly on crude meal or on their phenolic extracts obtained by methanolic extraction. Two types of meals were studied: (i) industrial (commercial) meals (I-meals), produced by a process that includes cooking at 95–100 °C and steam desolventizing at 105–107 °C, and (ii) non-industrial meals (NI-meals) obtained at pilot-scale with much milder heat treatment, that offer a higher total phenolic content. CA release through hydrolysis of sunflower meal (SFM) was successfully achieved with A. niger type-B feruloyl esterase (AnFaeB) and chlorogenic acid esterase (ChlE). Maximal amount of free CA released was of 54.0 ± 1.1 to 59.8 ± 2.1 μmol/g defatted dry matter (DDM) from I-SFM (94–100% hydrolysis yield) against 42.0 ± 1.1 to 52.3 ± 0.2 μmol/g DDM (59–73% hydrolysis yield) from NI- SFM in which CA release was hampered by a phenolic oxidation side-reaction, seemingly due to meal endogenous polyphenol oxidase activities. AnFaeB and ChlE hydrolysis of phenolic extracts from NI-SFM increased the CA amount obtained to 55.0–68.1 μmol/g DDM (77–95% hydrolysis yield). In all cases, AnFaeB showed broader specificity towards SFM caffeoyl quinic acid isomers than ChlE. In particular, ChlE did not hydrolyze 3-O-caffeoylquinic acid. The maximal amount of free SA released by AnFaeA hydrolysis was 41.3 ± 0.3 μmol/g DDM from NI-SFM (50% hydrolysis yield) and 32.3 ± 0.4 μmol/g DDM from the phenolic extract (64% hydrolysis yield), with AnFaeA also having sinapine transesterification activity that led to the synthesis of 1,2-di-O-sinapoyl-β-D-glucose. Finally, of all the substrates tested for enzymatic hydrolysis in our conditions, I-RSM and NI-SFM extract showed the best compromise between initial total phenolic content, hydrolysis yields and amounts of CA/SA released.

Bioactive molecules derived from smoke and seaweed Ecklonia maxima showing phytohormone-like activity in Spinacia oleracea L.

Bioactive compounds such as karrikinolide (KAR1 from smoke) and eckol (from the seaweed Ecklonia maxima) show promising effects on several important crop plants. These plant growth-stimulating organic biomolecules, along with crude extracts (smoke-water and Kelpak® product prepared from Ecklonia maxima), were tested on spinach plants. Eckol sprayed at 10−6 M significantly increased all the growth and biochemical parameters examined compared to control spinach plants. All tested plant growth biostimulants significantly increased total chlorophyll, carotenoids and protein content of spinach leaves. The cytokinin profile of spinach plants was also determined. Cis-zeatin, dihydrozeatin and isopentenyladenine types of cytokinins were promoted by both smoke- and seaweed-based biostimulants. In comparison to the control plants, the level of free sinapic acid was greater in all spinach plants treated with these biostimulants. The application of these biostimulants can help spinach crop by improving growth, yield and nutritional quality; moreover, they are organic and cost-effective.

Enhancing stability of echium seed oil and beta-sitosterol by their coencapsulation by complex coacervation using different combinations of wall materials and crosslinkers

Intake of omega-3 fatty acids and phytosterols aids in the reduction of cholesterol and serum triglycerides. However, both fatty acids and phytosterols are susceptible to oxidation. This work coencapsulated echium oil (source of stearidonic and alpha-linolenic fatty acids) and beta-sitosterol (phytosterol) by complex coacervation using different combinations of wall materials, and sinapic acid (SA) and transglutaminase as crosslinkers. High encapsulation yields were obtained (29–93% for SA; 68–100% for the mixture of oil and phytosterols) and retention of 49–99% and 16% for encapsulated and free SA, at 30 days-storage. Treatment with gelatin-arabic gum and 0.075 g SA/g gelatin showed the best results: 0.07 mg MDA/g capsule, and retention of 96, 90 and 74% for alpha-linolenic, stearidonic acid and beta-sitosterol at 30 days of storage, respectively. Thus, coencapsulation of echium oil and phytosterol using SA as the crosslinker was possible, obtaining effective vehicles for protection and application of these compounds in foods.

A Two-Step Bioconversion Process for Canolol Production from Rapeseed Meal Combining an Aspergillus niger Feruloyl Esterase and the Fungus Neolentinus lepideus.

Rapeseed meal is a cheap and abundant raw material, particularly rich in phenolic compounds of biotechnological interest. In this study, we developed a two-step bioconversion process of naturally occurring sinapic acid (4-hydroxy-3,5-dimethoxycinnamic acid) from rapeseed meal into canolol by combining the complementary potentialities of two filamentous fungi, the micromycete Aspergillus niger and the basidiomycete Neolentinus lepideus. Canolol could display numerous industrial applications because of its high antioxidant, antimutagenic and anticarcinogenic properties. In the first step of the process, the use of the enzyme feruloyl esterase type-A (named AnFaeA) produced with the recombinant strain A. niger BRFM451 made it possible to release free sinapic acid from the raw meal by hydrolysing the conjugated forms of sinapic acid in the meal (mainly sinapine and glucopyranosyl sinapate). An amount of 39 nkat AnFaeA per gram of raw meal, at 55 °C and pH 5, led to the recovery of 6.6 to 7.4 mg of free sinapic acid per gram raw meal, which corresponded to a global hydrolysis yield of 68 to 76% and a 100% hydrolysis of sinapine. Then, the XAD2 adsorbent (a styrene and divinylbenzene copolymer resin), used at pH 4, enabled the efficient recovery of the released sinapic acid, and its concentration after elution with ethanol. In the second step, 3-day-old submerged cultures of the strain N. lepideus BRFM15 were supplied with the recovered sinapic acid as the substrate of bioconversion into canolol by a non-oxidative decarboxylation pathway. Canolol production reached 1.3 g/L with a molar yield of bioconversion of 80% and a productivity of 100 mg/L day. The same XAD2 resin, when used at pH 7, allowed the recovery and purification of canolol from the culture broth of N. lepideus. The two-step process used mild conditions compatible with green chemistry.

Enzymatic synthesis of steryl hydroxycinnamates and their antioxidant activity

Steryl hydroxycinnamates are of increasing interest as they are antioxidant esters of phytosterols with potential cholesterol lowering properties. Apart from ferulates, also other plant steryl hydroxycinnamates have been identified in natural products. In this study hydroxycinnamic acid derivatives were ethylated enzymatically using Rhizomucor miehei lipase (yields from 16 to 97%), and transesterified by lipase from Candida rugosa to yield steryl hydroxycinnamates (yields from <LOQ to 55%). The influence of the structural differences between the hydroxycinnamic acid derivatives on the esterification yields was very different for the two lipases applied. Furthermore, the antioxidant activity of steryl and stearyl hydroxycinnamates was evaluated by DPPH radical scavenging activity and in two methyl linoleate systems. In bulk methyl linoleate, free sinapic acid showed the highest antioxidant activity over other sinapates, whereas in emulsified methyl linoleate, stearyl sinapate was highest. In conclusion, the enzymatic synthesis and antioxidant activity of steryl hydroxycinnamates is highly dependent on the acid structure.Practical applications: The enzymatic esterification of hydroxycinnamic acids is an environmentally friendly procedure to generate nonpolar hydroxycinnamates. These compounds have a broadened field of application due to their adapted polarity, and therefore potentially have an improved antioxidant activity in a wider range of food applications including multiphase systems. In the case of steryl hydroxycinnamates, the nonpolar part of the molecule brings additional health benefits, such as the cholesterol lowering properties, which makes them even more interesting as possible food additives with multiple technological and nutritional advantages.Hydroxycinnamic acids were ethylated enzymatically using R. miehei lipase, and transesterified by lipase from C. rugosa to yield steryl hydroxycinnamates. The influence of the structural differences between the hydroxycinnamic acids on the esterification yields was very different for the two lipases applied. Furthermore, the antioxidant activity of steryl and octadecyl hydroxycinnamates was evaluated by DPPH radical scavenging activity and in bulk and emulsified methyl linoleate system.

Self-nanoemulsifying drug delivery system of sinapic acid: In vitro and in vivo evaluation

Sinapic acid (SA) has been reported as a poorly soluble bioactive compound due to its dissolution rate and oral bioavailability, which are very poor. Therefore, in the current research work, different self-nanoemulsifying drug delivery systems (SNEDDS) of SA were developed for enhancement of its in vitro dissolution (drug release) and bioactivity. Different SA-loaded SNEDDS were prepared by low-energy emulsification methods, characterized and evaluated for various physicochemical and in vitro parameters such as thermodynamic stability, self-nanoemulsification efficiency, droplet size, polydispersity index (PDI), zeta potential (ZP), refractive index (RI), the % of transmittance (% T) and drug release profile. In vitro dissolution rate of SA was significantly enhanced from SNEDDS in comparison with SA suspension. The optimized SNEDDS of SA with droplet size of 12.4nm, PDI value of 0.158, ZP value of −32.8mV, RI value of 1.335, % T value of 98.7% and drug release profile of 96.8% was selected for in vitro antioxidant and in vivo anti-inflammatory studies. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay indicated significant antioxidant activity of optimized SA SNEDDS (IC50=4.52μg/ml) in comparison with free SA (IC50=7.62μg/ml) and standard ascorbic acid (IC50=40.78μg/ml). In vivo anti-inflammatory studies in rats also indicated that SA in optimized SNEDDS was significantly efficacious than SA suspension. Free SA as well as SA in SNEDDS was proved to be an excellent antioxidant in comparison with standard ascorbic acid. The results obtained in the current research work indicated the great potential of SNEDDS in the enhancement of in vitro dissolution rate and therapeutic efficacy of poorly soluble bioactive compounds such as SA.

Isolation and Identification of a Potent Radical Scavenger (Canolol) from Roasted High Erucic Mustard Seed Oil from Nepal and Its Formation during Roasting

Roasting of high erucic mustard (HEM) seed has been reported to give a typical flavor and increase the oxidative stability of the extracted oil. A potent radical scavenging compound was successfully isolated from roasted HEM seed oil in a single-step chromatographic separation using an amino solid-phase extraction column. Nuclear magnetic resonance and mass spectrometry spectra revealed the compound as 2,6-dimethoxy-4-vinylphenol (generally known as canolol), and its identity was fully confirmed by chemical synthesis. The formation of canolol during roasting was compared among HEM varieties (Brassica juncea, B. juncea var. oriental, Brassica nigra, and Sinapis alba) together with a low erucic rapeseed variety. HEM varieties were shown to produce less than one-third of canolol compared to rapeseed at similar roasting conditions. This observation was linked to a lower free sinapic acid content together with a lower loss of sinapic acid derivatives in the HEM varieties compared to rapeseed. Around 50% of the canolol formed in the roasted seed was shown to be extracted in the oil. Roasting of HEM seed before oil extraction was found to be a beneficial step to obtain canolol-enriched oil, which could improve the oxidative stability.

De-oiled rapeseed and a protein isolate: characterization of sinapic acid derivatives by HPLC–DAD and LC–MS

De-oiled rapeseed is a rich source of proteins and phenolic compounds. The phenolic compounds, namely sinapic acid derivatives (SAD), could occur as free sinapic acid, esterified (as sinapine, the choline ester of sinapic acid) and decarboxylated (as canolol) forms. Rapeseed protein preparations containing very low phenolic compounds have been the focus of our ongoing research. A precipitated rapeseed protein isolate is investigated for SAD such as sinapine, sinapoyl glucose, canolol using HPLC–DAD and LC–MS. Profile of the phenolic compounds of de-oiled rapeseed, press cakes and the precipitated protein isolate are compared. HPLC–DAD analysis indicated SAD; particularly sinapine is the main phenolic compound of all the substrates. The protein derivation process did not remarkably alter the profile of the investigated protein isolate.

Activities of Arabidopsis sinapoylglucose:malate sinapoyltransferase shed light on functional diversification of serine carboxypeptidase-like acyltransferases

Analysis of the catalytic properties of the serine carboxypeptidase-like (SCPL) 1- O-sinapoyl-β-glucose: l-malate sinapoyltransferase (SMT) from Arabidopsis showed that the enzyme exhibits besides its primary sinapoylation of l-malate, minor hydrolytic and disproportionation activities, producing free sinapic acid and 1,2-di- O-sinapoyl-β-glucose, respectively. The ability of the enzyme to liberate sinapic acid from the donor molecule 1- O-sinapoyl-β-glucose indicates the existence of a short-lived acylenzyme intermediate in the proposed random sequential bi–bi mechanism of catalysis. SMT-catalyzed formation of disinapoylglucose has been corroborated by docking studies with an established homology structure model that illustrates the possible binding of two 1- O-sinapoyl-β-glucose molecules in the active site and the intermolecular reaction of the two glucose esters. The SMT gene is embedded in a tandem cluster of five SCPL sinapoyltransferase genes, which encode enzymes with high amino acid sequence identities and partially overlapping substrate specificities. We assume that in recent duplications of genes encoding SCPL proteins, neofunctionalization of the duplicates to accept 1- O-sinapoyl-β-glucose as acyl donor was gained first, followed by subfunctionalization leading to different acyl acceptor specificities.

Antioxidant activity of rapeseed phenolics and their interactions with tocopherols during lipid oxidation

AbstractCommercial rapeseed press cakes are rich sources of phenolic compounds, namely, sinapic acid derivatives, which can be extracted as free sinapic acid and its bound forms (such as sinapine, the choline ester of sinapic acid). Fractionated rapeseed extracts rich in sinapic acid and sinapine were compared for their capacity to inhibit the formation of lipid oxidation products. Oxidation at 40°C was monitored by the formation of hydroperoxides (indicating primary oxidation products) and propanal (secondary oxidation products). The 70% methanolic extract of rapeseed meal, added as an equivalent of 500 μmol/kg oil (based on sinapic acid equivalent for sinapic acid‐rich extracts or sinapine equivalent for sinapinerich extracts) showed good antioxidative activity compared with the addition of 500 μmol/kg oil sinapic acid. Apart from this, the interaction between a mixture of α‐/γ‐tocopherol and sinapic acid was investigated using response surface methodology for the experimental design. The experiments indicated that the addition of sinapic acid (concentration dependent) caused inhibition of peroxide formation, complementing further lower endogenous tocopherol concentration in oils.

Antioxidative effect of the main sinapic acid derivatives from rapeseed and mustard oil by‐products

AbstractAmongst oilseeds, rapeseed and mustard are rich sources of phenolic compounds, which also prominent in the by‐products of their respective oil processing or in commercial rapeseed and mustard press cakes. These cakes are rich sources of sinapic acid derivatives, which could be extracted as free sinapic acid or sinapine, the choline ester of sinapic acid. Sinapic acid is a widely investigated antioxidative compound. However, the main compound in the press cakes is present as sinapine. Investigations on the free‐radical‐scavenging activity of sinapic acid and sinapine indicate that sinapine had a significant but lower activity as compared to sinapic acid. Apart from this, sinapic acid, sinapine and different tocopherols were compared as antioxidants for inhibition of the formation of lipid oxidation products in purified rapeseed oils. The oxidation at 40 °C was monitored by the formation of hydroperoxides and propanal. The experiments indicate that in contrast to tocopherol mixtures addition of sinapic acid causes increasing inhibition of hydroperoxide formation when enhancing the concentration from 50 to 500 μmol/kg oil. Sinapine was not able to inhibit the formation of hydroperoxides, compared to sinapic acid. This indicates that sinapic acid‐rich extracts, as compared to sinapine‐rich fractions, could better inhibit the lipid oxidation in bulk lipid systems.

Antioxidant Activity of the Main Phenolic Compounds Isolated from Hot Pepper Fruit (Capsicum annuum L.)

Four cultivars (Bronowicka Ostra, Cyklon, Tornado, and Tajfun) of pepper fruit Capsicum annuum L. were studied for phenolics contents and antioxidant activity. Two fractions of phenolics, flavonoids (with phenolic acids) and capsaicinoids, were isolated from the pericarp of pepper fruit at two growth stages (green and red) and were studied for their antioxidant capacity. Both fractions from red fruits had higher activities than those from green fruits. A comparison of the capsaicinoid fraction with the flavonoid and phenolic acid fraction from red fruit with respect to their antioxidant activity gave similar results. Phenolic compounds were separated and quantified by LC and HPLC. Contents of nine compounds were determined in the flavonoid and phenolic acid fraction: trans-p-feruloyl-beta-d-glucopyranoside, trans-p-sinapoyl-beta-d-glucopyranoside, quercetin 3-O-alpha-l-rhamnopyranoside-7-O-beta-d-glucopyranoside, trans-p-ferulyl alcohol-4-O-[6-(2-methyl-3-hydroxypropionyl] glucopyranoside, luteolin 6-C-beta-d-glucopyranoside-8-C-alpha-l-arabinopyranoside, apigenin 6-C-beta-d-glucopyranoside-8-C-alpha-l-arabinopyranoside, lutoeolin 7-O-[2-(beta-d-apiofuranosyl)-beta-d-glucopyranoside], quercetin 3-O-alpha-l-rhamnopyranoside, and luteolin 7-O-[2-(beta-d-apiofuranosyl)-4-(beta-d-glucopyranosyl)-6-malonyl]-beta-d-glucopyranoside. The main compounds of this fraction isolated from red pepper were sinapoyl and feruloyl glycosides, and the main compound from green pepper was quercetin-3-O-l-rhamnoside. Capsaicin and dihydrocapsaicin were the main components of the capsaicinoid fraction. A high correlation was found between the content of these compounds and the antioxidant activity of both fractions. Their antioxidant activities were elucidated by heat-induced oxidation in the beta-carotene-linoleic acid system and the antiradical activity by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) decoloration test. The highest antioxidant activity in the beta-carotene-linoleic acid system was found for trans-p-sinapoyl-beta-d-glucopyranoside, which was lower than the activity of free sinapic acid. Quercetin 3-O-alpha-l-rhamnopyranoside had the highest antiradical activity in the DPPH system, which was comparable to the activity of quercetin. The activities of capsaicin and dihydrocapsaicin were similar to that of trans-p-feruloyl-beta-d-glucopyranoside in the DPPH model system.

Sinapate dehydrodimers and sinapate-ferulate heterodimers in cereal dietary fiber.

Two 8-8-coupled sinapic acid dehydrodimers and at least three sinapate-ferulate heterodimers have been identified as saponification products from different insoluble and soluble cereal grain dietary fibers. The two 8-8-disinapates were authenticated by comparison of their GC retention times and mass spectra with authentic dehydrodimers synthesized from methyl or ethyl sinapate using two different single-electron metal oxidant systems. The highest amounts (481 microg/g) were found in wild rice insoluble dietary fiber. Model reactions showed that it is unlikely that the dehydrodisinapates detected are artifacts formed from free sinapic acid during the saponification procedure. The dehydrodisinapates presumably derive from radical coupling of sinapate-polymer esters in the cell wall; the radical coupling origin is further confirmed by finding 8-8 and 8-5 (and possibly 8-O-4) sinapate-ferulate cross-products. Sinapates therefore appear to have an analogous role to ferulates in cross-linking polysaccharides in cereal grains and presumably grass cell walls in general.

Carbohydrate and phenolic constituents in a comprehensive range of rapeseed and canola fractions: nutritional significance for animals.

AbstractCotyledons from 11 varieties of rapeseed and canola seed were extracted with hexane and analysed for tannins, α‐amylase inhibition value, starch and available carbohydrate by conventional methods. The sinapine content (total sinapic acid esters plus free sinapic acid) was determined by a spectrophotometric method. Samples of soya bean cotyledons, high‐tannin sorghum grain and commercial rapeseed and canola meals were also analysed for comparison. Sinapine in rapeseed and canola cotyledons was measured at mean levels of 2.67 and 2.85% respectively. Tannins were detected in these cotyledons but only at low levels. The α‐amylase inhibition values were also low or negative. Starch was detected in negligible amounts, and available carbohydrate at 13–14%. The predicted metabolisable energy (ME) value of commercial rapeseed or canola meal for poultry was found to be numerically close to the determined value when the available carbohydrate value was reduced to 35% of the observed value.

Enzymatic Synthesis of 1-Sinapoylglucose from Free Sinapic Acid and UDP-Glucose by a Cell-Free System from Raphanus sativus Seedlings

Abstract Protein extracts from seedlings of Raphanus sativus catalyze the transfer of the glucosyl moiety of UDP-glucose to the carboxyl group of phenolic acids. Enzymatic activity was determined spectrophotometrically by measuring the increase in absorbance at 360 nm and/or by the aid of high performance liquid chromatography (HPLC). From 12 phenolic acids tested as acceptors, sinapic acid was by far the best substrate. The glucosyltransfer to sinapic acid has a pH optimum near 7 and requires as SH group for activity, p-Chloromercuribenzoate (PCMB) inhibits activity, which can be restored by the addition of dithiothreitol (DTT). The formation of 1-sinapoylglucose was found to be a reversible reaction, since the addition of UDP results in a breakdown of the ester.

Metabolism of sinapic acid and related compounds in the rat.

1. Administration of sinapic acid to the rat results in the excretion of 3-hydroxy-5-methoxyphenylpropionic acid, dihydrosinapic acid, 3-hydroxy-5-methoxycinnamic acid and unchanged sinapic acid in the urine. The sinapic acid conjugate sinalbin is also catabolized to free sinapic acid and 3-hydroxy-5-methoxyphenylpropionic acid in the rat. 2. 3,4,5-Trimethoxycinnamic acid is metabolized in part to sinapic acid and 3-hydroxy-5-methoxyphenylpropionic acid. 3. 3,5-Dimethoxycinnamic acid is metabolized to 3-hydroxy-5-methoxycinnamic acid and 3-hydroxy-5-methoxyphenylpropionic acid. 4. The metabolic interrelationships of these compounds were studied by the administration of intermediates and a metabolic pathway is proposed. 5. The metabolism of the corresponding benzoic acids was studied, but these compounds and their metabolites were shown not to be intermediates or products of the metabolism of the related cinnamic acids.