Triticale Bran Research Articles

Modifications and in vitro absorption of 5-heptadecyresorcinol from cereals using digestion and ussing chamber models

5-Heptadecylresorcinol (AR-C17), a homologue of alkylresorcinols (ARs) and mainly observed in cereal brans, has stronger physiological functions compared with its homologues. However, not only is its content rare but also the purification low. Besides, few researches on its digestion characteristics and bioavailability limits its maximum applications. Here, we mainly relied on solid-state fermentation, embedment, in vitro models to systematically evaluate processing technologies, digestion and absorption characteristics of AR-C17. We showed that the highest content of AR-C17 was 57.6 μg/g extracted from triticale bran fermented by Saccharomyces cerevisiae relying on ultrasound-assistance. Additionally, AR-C17 was chiefly absorbed in duodenum and jejunum, and its apparent absorption increased by around 2.1 times when quercetin was added as the synergistic agent, which was higher than other phenolics in bran extract. Furthermore, AR-C17 embedded by β-cyclodextrin avoided the decomposition of in strong acidic environment, enhancing the retention rate to 96 % in in vitro digestion. According to the results above, we mixed AR-C17 with the quercetin, and embedded the mixture by β-cyclodextrin, which maximized the apparent absorption of AR-C17, reaching 19.8 % when the ratio of quercetin and AR-C17 was 1:1.

Quercetin Enhances the Availability of 5-Heptadecylresorcinol by Inhibiting the Expression of P-gp.

5-Heptadecylresorcinol (AR-C17), as the most important active monomer, is found in large quantities in wheat and triticale and plays a variety of health benefits, such as antidiabetic, anti-inflammatory, and antitumor. However, the low bioavailability of AR-C17 due to its low water solubility restricts its application. Moreover, the transport mechanism of AR-C17 is not fully understood. Here, we showed that the transport of AR-C17 in vitro was time- and concentration-dependent, and relatively higher temperature and lower pH obviously promoted the transport of AR-C17. Besides, transporters, especially P-glycoprotein (P-gp), markedly affected the transport of AR-C17 as well. Quercetin, a natural synergist in triticale bran (TB), was confirmed as an inhibitor of P-gp to promote the transport of AR-C17 in this study, and the bioavailability of AR-C17 reached the highest when the concentration ratio of quercetin to AR-C17 was 1:1.

Triticale flour in bakery and rusk products.

This paper researches the properties of two triticale varieties and three selection lines developed in Bashkortostan. The organoleptic indicators were typical of a standard grain; all the samples had a high protein content level. Gluten weight ratio matched the grain genotype. The falling number of the studied samples was low. Hydrothermal processing modes for triticale grain before grinding were optimized. The studied varieties and lines were found to yield 65.15%-70.18% of the flour. The conducted polynomial two-factor regression analysis proved that the flour yield factor is the grain softening period lasting 6 h. The dough deformation energy of the obtained flour samples was 67-129 units, the falling number 104-155 s indicates the low baking properties of the flour, thereby it was suggested to make brittle and crisp bakery products as crispbread and breadsticks. A breadstick formula with 60% wheat flour replaced with triticale flour was developed. The number of spore Rope bacteria increased slightly during the shelf life but did not exceed the permissible values. There have been developed formulas and methods to make crispbreads for producing crackers with triticale bran of 40% have been developed.

Cross-linking treatment of arabinoxylan improves its antioxidant and hypoglycemic activities after simulated in vitro digestion

The objective of the present study was to evaluate the impact of cross-linking treatment of arabinoxylan (AX) on its structure and physiological functions prior to and after the in vitro gastrointestinal digestion. The AX extracted from triticale bran was cross-linked using laccase. The reaction was monitored spectrophotometrically and verified based on the decrease in the ferulic acid content and an increase in the molecular weight. In comparison to the non-cross-linked AX (A-AX), the cross-linked AX (C-AX) exhibited a higher substitution degree, a more compact microstructure, and higher viscosity in the results of FT-IR, SEM, and rheological tests. The hydroxyl radical scavenging ability and the metal ion chelating ability of C-AX were higher than those of A-AX. After passing through the gastrointestinal tract simulated using an in vitro digestion model, in addition to the retentive larger molecular weight and network structure, the antioxidant activity and the glucosidase inhibition activity of C-AX were higher than those of A-AX. In conclusion, the cross-linking treatment of AX increased its tolerance of the extreme environment during the digestive process by modulating its structural characteristics.

Effect of bran addition on rheological properties of dough and quality of triticale bread

The purpose of this study was to examine the effects of the addition of triticale bran on the production of triticale bread. This work assessed in detail the properties of the dough and the quality of the bread with added bran, which was obtained from the grain of two selected triticale cultivars: Fredro and Panteon. The results showed that flours of these cultivars were suitable for bread-making. The addition of bran to baking blends positively affected the rheological properties of the dough and led to a substantial improvement in dough softening. Breads from the tested cultivars showed similar loaf volume. With the increasing addition of bran, breads of smaller volume and harder crumb were baked. In conclusion, the obtained results showed that triticale flour, and particularly its flour-bran blends with the addition of 5 and 10% of bran, resulted in the production of a new type of good quality bread. Practical applications Enrichment of triticale flour by adding a portion of bran allows such a flour to be used in baking of a good quality bread. According to our results, both of the examined triticale cultivars—Fredro and Panteon—can be considered for baking purposes. The addition of bran to baking blends at ratios of 5 and 10% to the flour mass had a beneficial effect on the rheological properties of the dough. There was a significant improvement in the softening of the dough prepared from triticale flour blends with bran compared to the value of this parameter in the dough without bran. Such an effect, of the use of bran in the blends is worthy of mention, because triticale doughs are most often characterized by high susceptibility to softening. The obtained results showed that triticale flour, and particularly its flour-bran blends with the addition of 5 and 10% of bran, resulted in the production of bread of good quality.

Interactive effects of molecular weight and degree of substitution on biological activities of arabinoxylan and its hydrolysates from triticale bran

Arabinoxylan (AX) has many beneficial health effects that are closely related to its structural characteristics. The current study aimed to investigate the effects of molecular weight (Mw) and degree of substitution (DS) on the antioxidant and hypoglycemic activities of triticale bran AX and its hydrolysates in vitro. At low and similar Mw, the antioxidant activity of AX was inversely proportional to its DS. When DS was close, the antioxidant activity of AX was inversely proportional to its Mw at high DS, but the opposite result was found at low DS. As for the hypoglycemic performance, when DS was similar, the hypoglycemic activity of AX was proportional to its Mw. At low and similar Mw, the α-glucosidase inhibitory ability and glucose adsorption ability of AX was positively correlated with DS, whereas the α-amylase inhibitory ability and glucose delayed absorption ability showed the opposite results. Mw and DS had significant effects on the antioxidant and hypoglycemic activities of AX, and these two factors often need to be combined to explain the varied effects under different conditions.

Lignans in triticale grain and triticale products

This study determined the lignan content of seven varieties of triticale grain and compared these values with wheat and rye grain, with triticale milling fractions (bran, shorts, and refined flour), and triticale products (sprouted grain, malt, fermented bread dough, baked dough, dried pasta and cooked pasta). Acid hydrolysis and enzymatic hydrolysis were used to release the lignans in extracts. We carried out qualitative and quantitative analysis of secoisolariciresinol, matairesinol, lariciresinol, syringaresinol, and pinoresinol levels using an Acquity UPLC liquid chromatograph with photodiode and mass Acquity TQD detectors. The most abundant lignan in all samples was syringaresinol, which made up 80% of all lignans. The lignan content of triticale grain varied from 392 to 792 μg·100gd·m−1 (acid hydrolysis) and from 685 to 1189 μg·100gd·m−1(enzymatic hydrolysis), depending on the variety. The lignan levels in triticale bran were almost three times those found in the grain; in flour, there was one fifth or even one tenth as much, whereas the shorts had similar levels as the grain. After three days of germination, the amount of lignans in the triticale grain increased by 17%–32%. A greater increase in lignan content (by factors of 1.4 and 2.3) was found in triticale after extrusion cooking. Fermentation led to lignan concentrations increasing slightly (2–14%), but this was then reduced in the baking process. Extracts from dry pasta contained similar or lower amounts than did the raw material, but the concentration was higher in cooked pasta (14%–47%). The enzymatic hydrolyzed extracts contained on average 36% higher lignan content than did the acid hydrolyzed samples.

Enrichment of maize and triticale bran with recombinant Aspergillus tubingensis ferulic acid esterase.

Ferulic acid is a natural antioxidant found in various plants and serves as a precursor for various fine chemicals, including the flavouring agent vanillin. However, expensive extraction methods have limited the commercial application of ferulic acid, in particular for the enrichment of food substrates. A recombinant Aspergillus tubingensis ferulic acid esterase Type A (FAEA) was expressed in Aspergillus niger D15#26 and purified with anion-exchange chromatography (3487U/mg, Km =0.43mM, Kcat=0.48/min on methyl ferulate). The 36-kDa AtFAEA protein showed maximum ferulic acid esterase activity at 50°C and pH 6, suggesting potential application in industrial processes. A crude AtFAEA preparation extracted 26.56 and 8.86mg/g ferulic acid from maize bran and triticale bran, respectively, and also significantly increased the levels of p-coumaric and caffeic acid from triticale bran. The cost-effective production of AtFAEA could therefore allow for the enrichment of brans generally used as food and fodder, or for the production of fine chemicals (such as ferulic and p-coumaric acid) from plant substrates. The potential for larger-scale production of AtFAEA was demonstrated with the A. niger D15[AtfaeA] strain yielding a higher enzyme activity (185.14 vs. 83.48U/ml) and volumetric productivity (3.86 vs. 1.74U/ml/h) in fed-batch than batch fermentation.

Triticale Bran Alkylresorcinols Enhance Resistance to Oxidative Stress in Mice Fed a High-Fat Diet.

Triticale (× Triticosecale Whitm.) is a cereal grain with high levels of alkyresorcinols (AR) concentrated in the bran. These phenolic lipids have been shown to reduce or inhibit triglyceride accumulation and protect against oxidation; however, their biological effects have yet to be evaluated in vivo. The purpose of this study was to determine the effects of ARs extracted from triticale bran (TB) added to a high–fat diet on the development of obesity and oxidative stress. CF-1 mice were fed a standard low-fat (LF) diet, a 60% high-fat diet (HF) and HF diets containing either 0.5% AR extract (HF-AR), 10% TB (HF-TB), or 0.5% vitamin E (HF-VE). Energy intake, weight gain, glucose tolerance, fasting blood glucose (FBG) levels, and body composition were determined. Oxygen radical absorbance capacity (ORAC), superoxide dismutase (SOD) activity, and glutathione (GSH) assays were performed on mice liver and heart tissues. The findings suggest that ARs may serve as a preventative measure against risks of oxidative damage associated with high-fat diets and obesity through their application as functional foods and neutraceuticals. Future studies aim to identify the in vivo mechanisms of action of ARs and the individual homologs involved in their favorable biological effects.

Chemical variability and antifungal activity of phenolic lipids from several Gramineae

An important class of phenolic lipids is alkylresorcinol (AR)-related compounds, which have been identified in many plants, fungi taxa, bacteria and some animals. Furthermore, there are several studies indicating that AR exhibit potent antimicrobial activity in plant defense possibly by their amphiphilic nature. This type of compounds can be found in large proportions in cereal grains such as wheat, rye, triticale and barley, whose occurrence is mainly located within the outer cuticle of the seed. As part of our research on antifungal compounds, the chemical profile and antifungal activity of extracts obtained from a set of 21 grasses were investigated. The chemical profile was determined by measuring total AR content (TAC) using Fast Blue RR protocol and by LC-DAD and LC-MS analysis of the resulting extracts for comparative drives by chemometrics through principal component analysis (PCA). TAC values indicated that triticale bran and wheat bran had the highest AR amounts with 45.0 and 33.1 mg AR/g dried material, respectively. In addition, the acetone-soluble extracts were evaluated in vitro against Fusarium oxysporum. The highest inhibition percentage against F. oxysporum was found for triticale bran-derived extract (81.9 and 76.9% at 10.0 and 1.0 µg/mL, respectively). The LC-DAD and LC-MS profiling for the 21 extracts led us to identify the some AR, involving AR with alkyl and alkenyl side chains possessing 13 – 19 carbons (tetradecenyl and hydroxytetradecenylresorcinol as main AR). Multivariate analysis (PCA) was performed on chromatographic data for all extracts. PCA-derived score plot showed that there is a relationship between the content of ARs and the antifungal activity. A bioguided fractionation leading to the isolation of the active AR is currently in the course.

Overexpression of Aspergillus tubingensis faeA in protease-deficient Aspergillus niger enables ferulic acid production from plant material

The production of ferulic acid esterase involved in the release of ferulic acid side groups from xylan was investigated in strains of Aspergillus tubingensis, Aspergillus carneus, Aspergillus niger and Rhizopus oryzae. The highest activity on triticale bran as sole carbon source was observed with the A. tubingensis T8.4 strain, which produced a type A ferulic acid esterase active against methyl p-coumarate, methyl ferulate and methyl sinapate. The activity of the A. tubingensis ferulic acid esterase (AtFAEA) was inhibited twofold by glucose and induced twofold in the presence of maize bran. An initial accumulation of endoglucanase was followed by the production of endoxylanase, suggesting a combined action with ferulic acid esterase on maize bran. A genomic copy of the A. tubingensis faeA gene was cloned and expressed in A. niger D15#26 under the control of the A. niger gpd promoter. The recombinant strain has reduced protease activity and does not acidify the media, therefore promoting high-level expression of recombinant enzymes. It produced 13.5 U/ml FAEA after 5 days on autoclaved maize bran as sole carbon source, which was threefold higher than for the A. tubingensis donor strain. The recombinant AtFAEA was able to extract 50 % of the available ferulic acid from non-pretreated maize bran, making this enzyme suitable for the biological production of ferulic acid from lignocellulosic plant material.

Cloning and characterization of the first GH10 and GH11 xylanases from Rhizopus oryzae.

The only available genome sequence for Rhizopus oryzae strain 99-880 was annotated to not encode any β-1,4-endoxylanase encoding genes of the glycoside hydrolase (GH) family 10 or 11. Here, we report the identification and cloning of two such members in R. oryzae strain NRRL 29086. Strain 29086 was one of several selected fungi grown on wheat or triticale bran and screened for xylanase activity among other hydrolytic actions. Its high activity (138 U/ml) in the culture supernatant led to the identification of two activity-stained proteins, designated Xyn-1 and Xyn-2 of respective molecular masses 32,000 and 22,000. These proteins were purified to electrophoretic homogeneity and characterized. The specific activities of Xyn-1 and Xyn-2 towards birchwood xylan were 605 and 7,710 U/mg, respectively. Kinetic data showed that the lower molecular weight Xyn-2 had a higher affinity (K m=3.2 ± 0.2 g/l) towards birchwood xylan than Xyn-1 by about 4-fold. The melting temperature (T m) of the two proteins, estimated to be in the range of 49.5-53.7 °C indicated that they are rather thermostable proteins. N-terminal and internal peptide sequences were obtained by chemical digestion of the purified xylanases to facilitate cloning, expression in Escherichia coli, and sequencing of the respective gene. The cloned Rhizopus xylanases were used to demonstrate release of xylose from flax shives-derived hemicellulose as model feedstock. Overall, this study expands the catalytic toolbox of GH10 and 11 family proteins that have applications in various industrial and bioproducts settings.

Determination of water-extractable polysaccharides in triticale bran

The content, composition, and molecular weight distribution (MWD) profile of water-extractable polysaccharides (WEP) in triticale (×Triticosecale Whitm.) bran were determined. Results were compared against wheat (Triticum sp.) and rye (Secale sp.) bran, as well as triticale straw and flakes. The effects of the following conditions on the extractability of sugars were compared and respective fractions obtained: (1) boiling water extraction, WEP-I; (2) successive enzyme treatment and dialysis, WEP-II; (3) successive ethanol fractionation, WEP-III. Although the highest yield of WEP was generally found in fraction II, fraction III had the least level of simple sugars as unwanted co-extractives. The extractability of xylose (Xyl) and arabinose (Ara) in all bran samples significantly increased (P<0.05) by up to 23.3% and 3.2% respectively, after precipitation with 80% ethanol. Amongst the cereal brans, wheat bran possessed a significantly higher (P<0.05) molar % of Ara (19.5%) and Xyl (29.6%), followed by triticale bran (14.5% Ara, 17.2% Xyl), and lowest in rye bran (7.2% Ara, 13.4% Xyl). Triticale bran (TB) had higher molecular weight (MW) polysaccharides and the widest MWD range (4400–401,000Da) of WEP, suggesting greater bioactive potential and high grain hardiness; prospects to be further investigated in future studies.

Responses to Sierra Rayne's Comments on “Optimization of alkylresorcinols extraction from triticale bran using response surface methodology [Agil et al. (2012), Food and Bioprocess Technology, 5(7), 2655–2664

1. FTIR gave us several lines since we had to run different samples with different concentrations to optimize in the range of relevant standards. 2. The software provided all lines in colour and were very narrow. Then, we selected the graphs of interests to be extracted from the instrument and applied them to PowerPoint, only the colour and thickness of the lines were changed. This is a very common way for any publication (e.g. presentation of GC or HPLC chromatograms) and does not mean that the data was manipulated. It was done simply to present the data more clearly for readers. Factors that affected alkylresorcinols spectra between 3000 and 2900:

Comment on “Optimization of Alkylresorcinols Extraction from Triticale Bran Using Response Surface Methodology [Agil et al. (2012), Food and Bioprocess Technology, 5(7), 2655–2664

Comment on “Optimization of Alkylresorcinols Extraction from Triticale Bran Using Response Surface Methodology [Agil et al. (2012), Food and Bioprocess Technology, 5(7), 2655–2664

Triticale bran: A novel dietary source of prebiotics and antioxidants in fermented dairy products

Triticale bran: A novel dietary source of prebiotics and antioxidants in fermented dairy products

Dual Functionality of Triticale as a Novel Dietary Source of Prebiotics with Antioxidant Activity in Fermented Dairy Products

The objectives of this study were to: (i) define the optimum concentration of triticale bran (TB) that can be incorporated in yogurt, (ii) evaluate the prebiotic effects of TB on microbial viability, pH and total titratable acidity (TTA) in yogurt across 28 days of cold storage, and (iii) measure the oxygen radical absorbance capacity (ORAC) of water-extractable polysaccharides (WEP) in TB. Lactobacillus bulgaricus and Streptococcus thermophilus were used as starter cultures. Lactobacillus acidophilus and Bifidobacterium lactis were used as probiotics. A concentration of 4% TB in yogurt was determined to be the maximum amount that could be added without causing synersis. By day 7, the number of bacteria greatly increased in yogurt samples containing TB and maintained higher viable bacteria counts at the end of the cold storage period, in comparison to controls (P ≤ 0.05). Confirming this data was the lower pH levels and higher TTA values of TB yogurt samples exhibited throughout 28 days (P ≤ 0.05). Polysaccharide extracts of TB exhibited strong antioxidant activity with an ORAC value of 33.86 ± 2.30 μmol trolox equivalents (TE)/g of bran. Results of this study suggest that TB may serve as a new prebiotic and antioxidant source for functional foods and nutraceutical applications.

Alternaria alternata as a new fungal enzyme system for the release of phenolic acids from wheat and triticale brans

This study describes the release of antioxidant ferulic acid from wheat and triticale brans by mixtures of extracellular enzymes produced in culture by a strain FC007 of Alternaria alternata, a dark mold originally isolated from Canadian wood log. The genus of the mold was confirmed as Alternaria by 18S ribosomal DNA characterization. Enzyme activities for feruloyl esterase (FAE) and polysaccharide hydrolyzing enzymes were measured, and conditions for release of ferulic acid and reducing sugars from the mentioned brans were evaluated. The highest level of FAE activity (89±7mUml(-1) fermentation culture) was obtained on the fifth day of fermentation on wheat bran as growth substrate. Depending on biomass and processing condition, up to 91.2 or 72.3% of the ferulic acid was released from wheat bran and triticale bran, respectively, indicating the proficiency of A. alternata extracellular enzymes in plant cell wall deconstruction. The apparent high extraction of ferulic acid from wheat and triticale brans represents a potential advantage of using a whole fungal cell enzyme complement over yields reported previously through an artificial assembly of cloned FAE with a particular xylanase in a cocktail format.

Optimization of alkylresorcinols extraction from triticale bran using response surface methodology

Optimization conditions for alkylresorcinols (ARs) extraction from triticale bran were determined using response surface methodology. A central composite design was used to determine the effects of extraction temperature (degrees Celsius) and solid-to-solvent ratio (weight per volume) on yield of saturated, unsaturated and total ARs. Extraction of ARs was affected significantly (p value ≤ 0.05) by temperature and solid-to-solvent ratio on the yield of saturated, unsaturated and total ARs. The highest quantity of total ARs from triticale bran was extracted at 24 °C between 16 and 24 h at a solid-to-solvent ratio (weight per volume) of 1:40 and ranged from 278 to 308 mg/100 g, while saturated and unsaturated ARs were 163 to 225 mg/100 g and 22 to 29 mg/100 g, respectively. The ARs identified by high-performance liquid chromatography included: C15:0, C17:0, C19:0, C21:0, C23:0 and C25:0. Also some unsaturated analogues (12.4% to 14.0%) were found including C17:1, C19:1, C21:1 and C23:1. Analysis of variance (p value = 0.05) indicated that the response surface methodology (RSM) developed for saturated, unsaturated and total ARs were adequate and explained most of the variability (85% to 89%) with high coefficient of multiple determination (R2 = 0.89). The main effect of the response variable was quadratic rather than a linear function. Our results showed that RSM is a tool that is useful to optimise experimental conditions for the extraction of ARs.

New Feruloyl Esterases to Access Phenolic Acids from Grass Biomass

In the Sorangium cellulosum strain So ce56 genome, two putative esterase-encoding genes (loci sce1896 and sce8927) were cloned, expressed in Escherichia coli, and the resulting enzymes (designated ScFAE1 and ScFAE2) were used to assess the possible release of ferulic acid (FA) from triticale and wheat brans, and an aqueous fraction of steam-exploded wheat straw. The two polypeptides, sharing only 30% sequence identity, exhibit a typical catalytic Ser-Asp-His triad, a characteristic of α/β-hydrolase fold proteins. Both ScFAE1 (35 kDa) and ScFAE2 (34 kDa) were purified to apparent homogeneity and comparison of their kinetic parameters indicated an apparent higher affinity of ScFAE2 than ScFAE1 towards the various feruloyl substrates. This property was reflected by the observation that ScFAE2 was capable of yielding up to 85% of FA from destarched triticale bran. In the steam-exploded wheat sample, more than 85% yield of FA or p-coumaric acid was also effected by ScFAE2 without the decomposition of valuable chemical such as furfural. The two cloned FAEs represent the first of myxobacterial origin to be characterized and they are classified as new members of the type D family of FAEs.