小麦麸皮结构层拉伸力学试样的手工制备方法探讨

制备小麦麸皮结构层试样是开展其机械特性拉伸试验的前提条件。能否利用手工剥离方法实现中间层和糊粉层的完整分离是制备小麦麸皮结构层试样的关键步骤，并形成了“两层说”和“三层说”的观点。该研究结合小麦麸皮结构层试样的制备要求，从小麦麸皮的显微组织结构、几何尺寸特性、实际剥离情况、宏观机械特性和杨氏模量复合法则等方面探讨了手工分离中间层和糊粉层的可行性问题。结果表明：利用手工剥离方法获得完整而纯粹的中间层和糊粉层试样是困难的。同时，明确了“三层说”分离中间层和糊粉层的方法是刮削工艺而不是撕离工艺，导致试样之间互相残留细胞组织，合理解释了两种试样的厚度和机械特性较为接近的原因。基于“两层说”的小麦麸皮结构层拉伸力学试验验证了层合板复合材料混合定律的普适应，表明当前利用“三层说”制备的中间层和糊粉层试样所得的拉伸力学特性参数的准确性不高。以上研究解决了手工分离中间层和糊粉层存在的一些疑惑，为利用小麦麸皮结构层机械特性探究其超微粉碎机理、优化其超微粉碎性能提供指导。     

Bran Size Distribution at Milling and Mechanical and Biochemical Characterization of Common Wheat Grain Outer Layers: A Relationship Assessment

Mechanical properties of wheat grain outer layers from common wheat (Triticum aestivum L.) cultivars known to display distinct milling behavior were analyzed using uniaxial tension tests. Tensile modulus and strain to rupture of the tissues distinguished between the wheat cultivars. Values of strain to rupture were related to coarse bran size generated by grain milling, a characteristic that distinguishes the two hardness classes. As content of an aleurone marker in total or first break flour was also related to coarse bran size, extensibility of wheat grain outer layers' could be a key parameter to explain the observed tissue mechanical behavior and thus distribution of the aleurone layer content in flours. As tissue mechanical properties are generally linked to the cell wall biochemical composition and structure, analysis of the main wheat outer layers' cell wall compounds was undertaken to establish relationships with the differences observed in mechanical properties. No clear correlation could be found with one of the wheat outer layers' component but involvement of the outer layers' cell wall structure in the tissues behavior at milling was confirmed.     

麦麸结构层粉体的电特性研究

为了确定麦麸结构层粉体静电场分离的可行性,该文以高纯度麦麸糊粉层和外果皮粉体为原料,研究其介电性、导电性和电晕带电特性的差异。结果表明,麦麸粉体的介电常数随频率的增加而降低,最终趋于一致;当频率较低时,外果皮的相对介电常数明显低于糊粉层;当频率为300 MHz时,两者的介电常数之比最大达5.3;麦麸粉体的导电性差,糊粉层粉体的电阻率为2300MΩ/cm,为外果皮电阻率的5倍;经正电晕带电后,糊粉层粉体所获电荷量约为外果皮的1.7倍,具有明显差异;单位质量麦麸粉体所获电荷量随电晕电压的增加而增加,而随麦麸粉体的含水率和粒径的增大而减小。总之,麦麸结构层粉体的介电性、导电性和电晕带电性均存在明显差异,可以利用介电泳和电晕带电后的静电场分离技术对麦麸结构层进行分离纯化。     

Enzymatic oxidative treatments of wheat bran layers: effects on ferulic acid composition and mechanical properties

Enzymatic treatments known to induce the gelation of feruloylated arabinoxylans solutions were applied to tissue strips isolated from peripheral layers of wheat grain to tentatively produce in situ arabinoxylan reticulation. The treatments by horseradish peroxidase (HRP) and manganese dependent peroxidase (MnP) induced a dimerization of ferulic acid (FA) in wheat bran with concomitant decrease of arabinoxylan solubility. Similar results were obtained, but to a lesser extent, by simple incubation of bran strips in water, suggesting the action of endogenous peroxidases. The fact that these treatments proved to be ineffective on the isolated aleurone layer and pericarp suggested that dimerization occurred mostly at the aleurone-pericarp interface. In addition, the MnP system generated a consumption of monomer and dimer of ferulic acid in the pericarp, perhaps due to their incorporation into lignin. Micro-mechanical tests using DMTA were performed on isolated tissue strips and showed that oxidation of wheat bran increased their mechanical strength (increase of stress and strain to rupture).     

Prediction of relative tissue proportions in wheat mill streams by fourier transform mid-infrared spectroscopy

Fourier-transform mid-infrared (FTIR) spectroscopy was investigated as a method to quantify the relative wheat grain tissue proportion in milling fractions. Spectra were acquired with a FTIR spectrometer equipped with an attenuated total reflectance device on ground samples, and the relative tissue proportion was determined according to the biochemical marker methodology as the reference method. Partial least-squares models were developed independently to predict the amount of outer pericarp, aleurone layer, starchy endosperm, and an intermediate layer (made up of inner pericarp plus seed coat plus nucellar epidermis). Good quality of prediction was obtained regardless of the target tissue. The standard errors of prediction obtained for the outer pericarp, intermediate layer, aleurone layer, and starchy endosperm quantification were, respectively, 3.4, 1.3, 3.4, and 4.6%.     

FTIR and Raman Signatures of Wheat Grain Peripheral Tissues

Hand‐dissection of mature grains from three common wheat cultivars (Triticum aestivum L.: Caphorn, Crousty, and Recital) and one durum wheat (T. durum Desf., Ardente) was performed to obtain pure samples of aleurone layer, hyaline layer, outer pericarp, and a composite layer made up of testa+inner pericarp. Vibrational spectral signatures were collected on both sides of the layers by ATR‐FTIR and Raman spectroscopy. Spectra were compared with biochemical analysis on the same samples which allowed identification of specific composition patterns in each tissue regardless of the cultivar. Considering the low penetration depth of ATR‐FTIR signal, the cuticles were evidenced on the external sides of outer pericarp, hyaline layer, and testa. Spectra from testa of red and white wheats were clearly distinguished. FTIR spectroscopy, combined with statistical analysis, was successful in identifying the specific spectral signature for each peripheral tissue of wheat grains. In the 1500–800 cm^‐1 spectral region, multivariate models allowed accurate prediction of the histological origin of the pericarp, hyaline, and aleurone layers regardless of the analyzed side, and the testa but with a lower performance.     

Influence of Structural Characteristics of Aleurone Layer on Milling Behavior of Durum Wheat (Triticum durum Desf.)

The structure of the aleurone layer was considered for many years as a potential factor influencing wheat milling efficiency. Eight durum wheat samples of different milling values, including distinct cultivars and harvesting conditions, were employed to investigate the structural characteristics of the aleurone layer through image analysis of kernel sections. Particular attention was paid to tissue thickness and structural irregularity of its interface with the starchy endosperm. Wheat cultivar, agricultural conditions, and location of measurement within the grain had an influence similar to both thickness and irregularity of the aleurone layer. Conversely, grain weight and morphology showed no effect on these parameters. Statistical investigation demonstrated no correlation between structural characteristics and wheat milling behavior. However, the negative correlation between the extraction rate of semolina and starch content in the bran fraction, which was used as an indicator of the endosperm‐aleurone dissociation extent, demonstrated the relevance of the tissue adhesion on milling efficiency.     

Influence of UV Exposure on Phenolic Acid Content,Mechanical Properties of Bran,and Milling Behavior of Durum Wheat (Triticum Durum Desf.)

Durum wheat bran was exposed to UV radiation up to 48 hr and the changes in ferulic acid (FA) content in the peripheral part s of grain were measured. The treatment resulted in a 25% decrease in FA monomer and a 44% decrease in dehydrodiferulic acid (DHD) ester‐linked to the cell‐wall arabinoxylans. This reduction was partly explained by a significant increase of FA (30%) and DHD (36%) engaged in hot alkali‐labile linkages. The results suggest that UV irradiation induced the formation of new cross‐links between feruloylated arabinoxylan and lignin in the pericarp. The effects of UV treatment on bran mechanical properties and wheat milling behavior were investigated. UV irradiation for 15 hr increased the stress to rupture by 30% and decreased the extensibility of bran tissues by 54%. This stiffening was associated with an increase in bran friability during grinding. Although this effect was due in part to the hydrothermal history of the grain, chemical modification induced by UV significantly influenced the size reduction of bran particles, which can be explained by the modification of the mechanical properties of bran. Relationships between the organization of cell‐wall polymers, the mechanical properties of tissues, and the behavior of wheat grain during milling were investigated.     

Distribution of Peroxidases in Durum Wheat (Triticum durum)

A biochemical study of the main durum wheat milling fractions (bran, embryo, and semolina) showed that peroxidases (POD) were present in multiple forms in the kernel and appeared to be tissue specific: one form for the embryo, one for the endosperm, one for the subaleuronic layer, and one for the outer layers. Large varietal differences were found regarding both the composition and the POD activity. POD activity, detected by diaminobenzidine, was found mainly in the cell wall of the subaleurone layer and inside some specific, differentiated cells of the embryo. Immuno‐localization with antibodies of durum wheat POD showed the presence of POD in several layers of the pericarp (epidermis) and the seed coat (testa), in the embryo, and also in the endosperm. In this latter tissue, the staining intensity decreased gradually from the outer layers toward the center of the kernel. The localization of POD in durum wheat kernel suggests specific functions for different forms.     

Effect of Corn Milling Practices on Aleurone Layer Cells and Their Unique Phytosterols

Coarse and fine fiber fractions obtained from the corn wet‐milling processes, with and without steeping chemicals (SO₂ and lactic acid), were evaluated microscopically for structure and analytically for recovery of phytosterol compounds from the fiber oil. Microscopic results showed that wet milling, with and without chemicals during steeping, changed the line of fracture between pericarp and endosperm and therefore affected the recovery of the aleurone layer in coarse (pericarp) and fine (endosperm cellular structure) fiber. Analytical results showed that most of the phytosterols and mainly phytostanols in corn fiber are contributed by the aleurone layer. Hand‐dissection studies were performed to separate the two layers that comprise the wet‐milled coarse fiber, the aleurone, and pericarp layer. Analyses revealed that the aleurone contained 8× more phytosterols than the pericarp.     

Comparative Studies on Composition and Distribution of Phenolic Acids in Cereal Grain Botanical Fractions

The phenolic acid composition and concentration of four manually separated fractions (pericarp, aleurone layer, germ, and endosperm fractions) as well as whole grains of yellow corn, wheat, barley, and oats were analyzed by HPLC‐MS/MS following microwave‐assisted alkaline aqueous extraction. Phenolic acid compositions in whole grains and their fractions were similar, with minor differences among the grain fractions. Significant differences (P < 0.05), however, were observed in phenolic acid concentrations among cereal types, within cereal varieties, and among grain fractions, with yellow corn exhibiting the highest values. The concentrations of p‐coumaric and syringic acid in the pericarp were 10‐ to 15‐fold and 6‐ to 10‐fold higher, respectively, in yellow corn than in wheat, barley, and oats. In the aleurone layer, sinapic and vanillic acids in yellow corn were about 8‐ and 30‐fold more than in wheat. The germ fraction of wheat had 1.8 times more syringic acid than yellow corn germ. Grain fractions, excluding endosperm, had enhanced levels of phenolic acids compared with whole grain. Sinapic acid was more concentrated in the pericarp and germ of wheat, whereas isoferulic acid was concentrated in the germ of purple barley. Syringic and vanillic acids were concentrated in the pericarp and sinapic acid in the aleurone layer of yellow corn. These findings are important in understanding the composition and distribution of phenolic acids, and they act as a guide in identification of grain fractions for use as food ingredients. In addition, yellow corn fractions (aleurone and pericarp) may be potential alternative phenolic‐rich functional food ingredients in grain‐based food products.     

Evaluation of Degree of Elasticity and Other Mechanical Properties of Wheat Kernels

Three samples were selected representing bread, soft, and durum wheat. Uniaxial compression and stress relaxation tests were performed on wheat kernels. Force‐deformation curves from intact wheat grain typically exhibited at least two points of inflection (PI) at ≈0.1 and 0.2 mm displacement. The first PI is related to the mechanical properties of all the bran layers. The second PI (0.2 mm) seems to be the endosperm boundary near the aleurone layer. These structures had higher degree of elasticity (DE) compared to the inner endosperm (0.5–0.6 mm). Besides wheat class and specific structures of the caryopsis, moisture content is a prominent factor affecting the mechanical strength of kernels. Stress relaxation tests show that bread wheat kernels with 69.2% DE at 13% moisture decreased to 31.6% DE with additional 6% moisture content. Soft wheat kernels DE of 61.0% at 13% moisture decreased to 22.7% at 19.7% moisture. Stress relaxation revealed pronounced time‐dependence. However, the differences of stress values at 120–180 sec were not significant in all wheat classes and moisture contents evaluated. The stress values after 120 sec might be attributed to the elastic deformation of the kernels.     

Differences in the Aleurone Layer Fate Between Hard and Soft Common Wheats at Grain Milling

In the milling process, efficient separation between the starchy endosperm and the other grain tissues is a key parameter estimated by ash measurement. Because this separation occurs near the aleurone layer interface, better understanding of this tissue fractionation is critical for a better analysis of the wheat milling behavior. Samples from hard and soft common wheat cultivars that had the same protein content were processed on a pilot mill, and whole grain meals or flour streams were analyzed for ash content. The para‐coumaric acid (p‐CA) and phytic acid flour contents were compared with ash measurement and used as markers of the aleurone cell walls or aleurone cell content, respectively. A greater amount of phytic acid in hard wheat flour compared with soft wheat flour was found and reveals a distinct milling behavior between those wheat classes, mainly at the breaking step. Therefore simple ash content measurement is not sufficient to analyze flour purity. At the reduction stage, quantity of phytic acid increases with the other markers and may result from the overall mechanical resistance of the aleurone tissue. As a consequence, wheat hardness not only determines grain milling behavior but also affects flour composition.     

In‐Depth Study of Durum Wheat Grain Tissue Distribution at Milling

The starchy endosperm proportion in durum wheat grain and its ability to be isolated from the peripheral tissues appear as main intrinsic characteristics potentially related to the milling value but still difficult to assess. In this study, several durum wheat samples displaying distinct grading characteristics were analyzed and processed through a pilot mill. The histological composition of grains and milling fractions was monitored by using identified biochemical markers of each wheat grain tissue. Contrasted milling yields of semolina and flour were observed between samples, despite displaying a similar starchy endosperm proportion determined by hand dissection. These yields were related both to differences in the starchy endosperm extraction and to the presence of the aleurone layer, particularly its cellular content. Furthermore, two distinct types of fractionation behavior of the aleurone layer were distinguished depending on the wheat grain sample. Extraction of the envelopes and embryonic axis into semolina and flours were found negligible in comparison with the other tissues.     

Comparison of Swiss red wheat grain and fractions for their antioxidant properties

Swiss red wheat grain, bran, aleurone, and micronized aleurone were examined and compared for their free radical scavenging properties against 2,2-diphenyl-1-picrylhydrazyl radical (DPPH*), radical cation ABTS*+ and peroxide radical anion O(2)*-, oxygen radical absorbance capacity (ORAC), chelating capacity, total phenolic content (TPC), and phenolic acid composition. The results showed that micronized aleurone, aleurone, bran, and grain may significantly differ in their antioxidant properties, TPC, and phenolic acid composition. Micronized aleurone had the greatest antioxidant activities, TPC, and concentrations of all identified phenolic acids, suggesting the potential of postharvesting treatment on antioxidant activities and availability of TPC and phenolic acids. Ferulic acid was the predominant phenolic acid in Swiss red wheat and accounted for approximately 57-77% of total phenolic acids on a weight basis. Ferulic acid concentration was well correlated with scavenging activities against radical cation and superoxide anion, TPC, and other phenolic acid concentrations, suggesting the potential use of ferulic acid as a marker of wheat antioxidants. In addition, 50% acetone and ethanol were compared for their effects on wheat ORAC values. The ORAC value of 50% acetone extracts was 3-20-fold greater than that of the ethanol extracts, indicating that 50% acetone may be a better solvent system for monitoring antioxidant properties of wheat. These data suggest the possibility to improve the antioxidant release from wheat-based food ingredients through postharvesting treatment or processing.     

Formation of phenolic microbial metabolites and short-chain Fatty acids from rye, wheat, and oat bran and their fractions in the metabolical in vitro colon model

Rye bran and aleurone, wheat bran and aleurone, and oat bran and cell wall concentrate were compared in their in vitro gut fermentation patterns of individual phenolic acids and short-chain fatty acids, preceded by enzymatic in vitro digestion mimicking small intestinal events. The formation of phenolic metabolites was the most pronounced from the wheat aleurone fraction. Phenylpropionic acids, presumably derived from ferulic acid (FA), were the major phenyl metabolites formed from all bran preparations. The processed rye, wheat, and oat bran fractions contained more water-extractable dietary fiber (DF) and had smaller particle sizes and were thus more easily fermentable than the corresponding brans. Rye aleurone and bran had the highest fermentation rate and extent probably due to high fructan and water-extractable arabinoxylan content. Oat samples also had a high content of water-extractable DF, β-glucan, but their fermentation rate was lower. Enzymatic digestion prior to in vitro colon fermentation changed the structure of oat cell walls as visualized by microscopy and increased the particle size, which is suggested to have retarded the fermentability of oat samples. Wheat bran was the most slowly fermentable among the studied samples, presumably due to the high proportion of water-unextractable DF. The in vitro digestion reduced the fructan content of wheat samples, thus also decreasing their fermentability. Among the studied short-chain fatty acids, acetate dominated the profiles. The highest and lowest production of propionate was from the oat and wheat samples, respectively. Interestingly, wheat aleurone generated similar amounts of butyrate as the rye fractions even without rapid gas production.     

Structure, chemical composition, and xylanase degradation of external layers isolated from developing wheat grain

The external layers of wheat grain were investigated during maturation with respect to chemical and structural features and xylanase degradability. Cytochemical changes were observed in the isolated peripheral tissues of the wheat grain at four defined stages following anthesis. Marked chemical changes were highlighted at 11 days after anthesis, for which protein and lipid contents varied weakly. The profile of esterified ferulic acid showed large variation in the maturing peripheral layers of grain in contrast to the deposition of ferulate dimers, p-coumaric and sinapic acids. Lignin was monitored at the latest stages of ripening, which corresponds to the cessation of reserve accumulation in the grain. Arabinoxylans (AX) reached a maximum at 20 days and did not display any significant change in arabinosyl substitution proportion until ripeness. When submitted to xylanase, all outer layers were similarly altered in the proportion of soluble AX except for the peripheral tissues of the 11-day-aged wheat grain that had very little AX. Aleurone and nucellar layers were mostly degraded, whereas pericarp stayed intact at all stages of maturation. This degradation pattern was connected with the preferential immunolocalization of xylanase in aleurone and nucellar layers irrespective of the developmental stages. Further chemical examination of the enzyme-digested peripheral tissues of the grain supports the facts that ferulic ester is not a limiting factor in enzyme efficiency. Arabinose branching, ferulic dimers, and ether-linked monomers that are deposited early in the external layers would have more relevance to the in situ degradability of AX.     

Effect of Outer Bran Layers from Germinated Wheat Grains on Breadmaking Properties

Wheat grains were germinated in the dark at room temperature (24°C) for 1, 2, 3, 5, and 8 days. Germinated wheat grains were pearled in a Waring blender, and the outer bran layers were separated from the pearled grains. Breadmaking was performed with wheat flour and 10% of the outer bran layer. Breadmaking properties (bread height [mm] and specific volume [cm³/g]) were gradually enhanced by blending the germinated outer bran layers, and maximum specific volume was obtained after 5 days of germination. However, the improvement was lost after 8 days of germination. Rapid ViscoAnalyser (RVA) and Brabender Farinograph profiles of wheat flour and outer bran layers (10%) indicated that the maximum decrease of peak viscosity and increase of angle of tail were obtained after 3 and 5 days of germination, respectively. Those changes are presumed to be due to the action of enzymes found in the outer bran layers. Activities of α‐ and β‐amylase, lipase, protease, and xylanase in the outer bran layers were measured, and correlation coefficients (r) between breadmaking properties and peak viscosity (RVA), angle of tail (farinograph), and enzyme activities were calculated. These data suggest that xylanase and α‐amylase activities in the outer bran layers were highly related to the enhancement of the breadmaking properties.     

Distribution of mineral elements in the outer layer of rice and wheat grains,using electron microprobe X-Ray analysis

Electron microprobe X-ray analysis was used to determine the transversal microdistribution of P, K, Mg, Ca, Fe, and Mn, with special attention to the outer layers of rice and wheat grains. P, K, Mg, Fe, and Mn were concentrated in the aleurone layer in each case. In particular, P, Mg, and K were highly concentrated in the subcellular particles of the aleurone layer, and had very similar distribution patterns in the outer layers of the matured grain of rice and wheat. By contrast, Ca was abundant in the pericarp.     

Effect of Hydrolyzing Enzymes on Wheat Bran Cell Wall Integrity and Protein Solubility

Wheat bran contains good quality protein, but given its location inside aleurone cells, this protein has restricted digestibility. The aim of this work was to liberate and solubilize wheat bran proteins via cell wall degradation by using carbohydrate‐hydrolyzing and proteolytic enzymes without causing extensive protein hydrolysis. Bran incubated with water (without added enzymes) for 16 h increased the solubilized organic nitrogen content from 14.0 to 42.8%. Enzymes with solely carbohydrate‐hydrolyzing activity increased the water‐soluble pentosan and reducing sugar contents but did not significantly increase protein solubilization or protein release from the aleurone cells. Enzymes with proteolytic activity significantly increased the solubilization of protein to 58.2% already at 4 h. Significant protein hydrolysis was detected with a high dosage of protease. However, based on light microscopy, the enzymatic treatment mainly modified the proteins in the subaleurone layer, and it was less effective on proteins inside the aleurone cells. With optimized protease treatment (3 h, 35°C, and 550 nkat/g), effective protein solubilization (>48%) without extensive protein hydrolysis (free amino nitrogen content <45 mg/L) was achieved. In conclusion, intensive solubilization of proteins in the subaleurone layer of wheat bran is possible by using exogenous enzymes with proteolytic activities.