Arabinoxylan, ß-glucan and pectin in barley and malt endosperm cell walls: a microstructure study using CLSM and cryo-SEM.

The architecture of endosperm cell walls in Hordeum vulgare (barley) differs remarkably from that of other grass species and is affected by germination or malting. Here, the cell wall microstructure is investigated using (bio)chemical analyses, cryogenic scanning electron microscopy (cryo-SEM) and confocal laser scanning microscopy (CLSM) as the main techniques. The relative proportions of ß-glucan, arabinoxylan and pectin in cell walls were 61, 34 and 5%, respectively. The average thickness of a single endosperm cell wall was 0.30 µm, as estimated by the cryo-SEM analysis of barley seeds, which was reduced to 0.16 µm after malting. After fluorescent staining, 3D confocal multiphoton microscopy (multiphoton CLSM) imaging revealed the complex cell wall architecture. The endosperm cell wall is composed of a structure in which arabinoxylan and pectin are colocalized on the outside, with ß-glucan depositions on the inside. During germination, arabinoxylan and ß-glucan are hydrolysed, but unlike ß-glucan, arabinoxylan remains present in defined cell walls in malt. Integrating the results, an enhanced model for the endosperm cell walls in barley is proposed.

A simple method for analysis of vitamin A palmitate in fortified cereal products using direct solvent extraction followed by reversed-phase HPLC with UV detection.

Vitamin A is generally analysed using a time-consuming and possibly detrimental saponification step, followed by extraction and HPLC analysis. We here developed a new method to analyse retinyl palmitate (RP) (also known as vitamin A palmitate) without the need for saponification and validated it in model systems consisting of RP, soy oil and wheat bran, and in RP-fortified cereal products. Two direct solvent extraction protocols using acetone/methanol (7/3, v/v) or chloroform/methanol (1/1, v/v) were tested. After extraction, RP was quantified by reversed-phase HPLC with UV detection. The HPLC method had low limits of detection (0.01 µg/mL) and quantification (0.03 µg/mL). Both extraction protocols showed a good recovery (88-105 %) and intra-and inter-day precision (<5%) for RP extraction from the model systems. The obtained results corresponded to results obtained using a golden standard approach. For complex food matrices like bread and cookies, the chloroform/methanol extraction protocol showed the best performance characteristics.

Stabilization of Vitamin A by Cereal Bran: The Importance of the Balance between Antioxidants, Pro-oxidants, and Oxidation-Sensitive Components.

This study investigated the contribution of bran antioxidants and lipids to the stabilizing effect of cereal bran on vitamin A during accelerated storage. Hereto, wheat and rice bran samples subjected to a sequential extraction process were used. Vitamin A stabilization was more pronounced for wheat compared to rice bran. This was attributed to the higher antioxidant capacity and lower degree of lipid oxidation of wheat compared to rice bran. Removal of the chloroform/methanol-extractable fraction resulted in a substantial decrease in vitamin A retention from 78 to 26% for wheat bran and from 30 to 0% for rice bran after 2 weeks of accelerated storage. However, the vitamin A-stabilizing effect could not be attributed to specific components. The ability of cereal bran to stabilize vitamin A is therefore believed to be determined by the balance of antioxidants, pro-oxidants, and oxidation-sensitive components in the system.

Stabilisation of vitamin A by wheat bran is affected by wheat bran antioxidants, bound lipids and endogenous lipase activity.

Food fortification is an efficient strategy to combat vitamin A deficiency. However, the stability of vitamin A during storage is low. Cereal bran can be used as a natural and affordable stabilising agent, but the mechanism behind this stabilisation remains unclear. To unravel this mechanism, vitamin A stabilisation was studied during an accelerated storage experiment (60 °C, 70% relative humidity) using a set of 30 in-house modified wheat bran samples. The characteristics of these samples were linked to vitamin A stabilisation during storage using forward regression modelling. While all wheat bran samples could stabilise vitamin A to a significant extent, the stabilising effect was more pronounced for samples with a high antioxidant capacity, high bound lipid content and low lipase activity. The main effect of lipase activity was more than thrice as large as the main effects of antioxidant capacity and bound lipid content. These results suggest that wheat bran antioxidants and bound lipids protect vitamin A from degradation during storage, while endogenous lipase activity counteracts the stabilising effect. Based on these findings, modified wheat bran mixed with vitamin A can be a cost-effective and healthy aid in food fortification by providing high vitamin A stability.

Conversion of Retinyl Palmitate to Retinol by Wheat Bran Endogenous Lipase Reduces Vitamin A Stability.

Wheat bran can be used as a cost-effective food ingredient to stabilise vitamin A. However, wheat bran endogenous enzymes have been shown to reduce vitamin A stability. In this study, we elucidated the mechanism for this negative effect in an accelerated storage experiment with model systems consisting of native or toasted wheat bran, soy oil and retinyl palmitate (RP). Both native and toasted wheat bran substantially stabilised RP. While RP was entirely degraded after ten days of storage in the absence of wheat bran, the RP retention after ten days was 22 ± 2% and 75 ± 5% in the presence of native and toasted bran, respectively. The significantly stronger stabilising effect of toasted bran was attributed to the absence of bran endogenous enzymes. In contrast to toasted bran systems, noticeable free fatty acid production was observed for native bran systems. However, this did not result in a pronounced lipid oxidation. Next to lipid hydrolysis, wheat bran lipase was shown to hydrolyse retinyl esters to the less stable retinol and fatty acids. This reaction could explain the major part, about 66 ± 5%, of the difference in RP stabilisation between native and toasted wheat bran.

The Wheat Starchy Endosperm Protein Gradient as a Function of Cultivar and N-fertilization Is Reflected in Mill Stream Protein Content and Composition.

Within the wheat starchy endosperm, the protein content increases biexponentially from the inner to outer endosperm. Here, we studied how this protein gradient is reflected in mill fractions using three cultivars (Claire, Apache, and Akteur) grown without and with N-fertilization (300 kg N ha-1). The increasing protein content in successive break fractions was shown to reflect the protein gradient within the starchy endosperm. The increasing protein content in successive reduction fractions was primarily due to more aleurone contamination and protein-rich material being harder to reduce in particle size. The miller's bran fractions had the highest protein content because of their high sub-aleurone and aleurone content. Additionally, the break fractions were used to deepen our understanding of the protein composition gradient. The gradient in relative gluten content, increasing from inner to outer endosperm, was more pronounced without N-fertilization than with and reached levels up to 87.3%. Regarding the gluten composition gradient, no consistent trends were observed over cultivars when N-fertilization was applied. This could, at least partly, explain why there is no consensus on the gluten composition gradient in the literature. This study aids millers in managing fluctuations in the functionality of specific flour streams, producing specialized flours, and coping with lower-quality wheat.

Impact of process parameters on the specific volume of wholemeal wheat bread made using sourdough- and baker's yeast-based leavening strategies.

The final quality of wholemeal wheat bread is determined by the process parameter settings and leavening strategy. We hypothesise that the used leavening strategy may influence the optimal process parameter settings and, as such, the specific volume of the bread loaf. To analyse this interaction, bread was leavened with (i) a type 1 sourdough (SB), (ii) a type 1 sourdough combined with baker's yeast (YSB), or (iii) baker's yeast (YB). For each leavening strategy, the specific volume of bread, in response to variations in mixing time (4-10/4-14 min), water absorption (60-85 %), and proofing time (1-7/1-3 h), was analysed using an I-optimal response surface experimental design. Data modelling identified a substantially lower maximal specific volume of SB (2.13 mL/g), compared to YSB (3.30 mL/g) and YB (3.26 mL/g). The proofing time and water absorption mostly influenced the specific volume of the SB and YSB, respectively. However, the mixing and proofing times mainly affected the specific volume of YB. The type 1 sourdough reduced the mixing time and water absorption required for an optimal specific volume of bread compared to baker's yeast. These results challenge the idea of yielding higher volumes upon using sourdough compared to baker's yeast and highlight the importance of optimisation of bread dough formulations and breadmaking processes.

The Contribution of Sub-Aleurone Cells to Wheat Endosperm Protein Content and Gradient Is Dependent on Cultivar and N-Fertilization Level.

The proteins in the starchy endosperm of wheat determine wheat quality and exhibit a quantitative gradient decreasing from the outer to inner endosperm. Here, we investigate how protein-rich sub-aleurone cells contribute to the protein content and gradient by studying three cultivars, each cultivated at three levels of nitrogen (N)-fertilization. The observed increased protein content with increased N-fertilization was cultivar-dependent. Image analysis showed that the underlying protein gradient could be described by a declining biexponential curve, with protein contents up to 32.0% in the sub-aleurone. Cultivars did not differ in protein content in the center of the cheeks and only differed in the outer endosperm when N-fertilization is applied. N-Fertilization resulted in relatively higher increases in protein content in the outer compared to inner endosperm. Hence, sub-aleurone cells could affect the classification of cultivars by baking quality. Cultivar selection and N-fertilization could furthermore be promising techniques to produce protein-rich miller's bran.

Arabinoxylan from non-malted cereals can act as mouthfeel contributor in beer.

A satisfying mouthfeel is essential for the production of non-alcoholic, low-alcohol beers and light beers. This paper highlights the importance of non-starch carbohydrates as mouthfeel contributors in this context. Beers were brewed with a substitution of 20 % barley malt grits by non-malted barley, rye or oats compared to a control. For the beer brewed with rye, both a 53 % increase in arabinoxylan content and an increase in the average degree of polymerization from 29 to 50 were observed. Compared to the control beer (1.48 mm²/s), viscosity was the highest for the rye beer (1.85 mm²/s). Multivariate data analysis underlined the role of arabinoxylan content and degree of polymerization as determinants of beer viscosity. A sensory panel distinguished a low-alcohol rye beer as the one with increased fullness compared to a 100 % malt beer. These experiments suggest that rye addition can be used as a strategy to increase the beer fullness.

Different gelatinization characteristics of small and large barley starch granules impact their enzymatic hydrolysis and sugar production during mashing.

This study investigates the impact of different gelatinization characteristics of small and large barley starch granules on their enzymatic hydrolysis and sugar production during mashing. Therefore, a barley malt suspension was consecutively incubated at 45, 62, 72 and 78 °C to monitor starch behavior and enzymatic starch hydrolysis and sugar production. The combination of microscopic and rapid visco analyses showed that small starch granules persisted longer in the system and were present non-gelatinized at temperatures higher than 62 °C. HPAEC-PAD analysis showed that 8% of the total amount of starch, predominantly small granules, gelatinized at temperatures between 62 °C and 78 °C. Due to their delayed gelatinization in this system, their enzymatic hydrolysis yielded relatively more dextrins compared to what was observed for large granules. It was concluded that small granules should be taken into account when optimizing enzymatic hydrolysis of barley starch, like in brewing, distilling or bio-ethanol production.