Enzyme pre-milling treatments improved milling performance of chickpeas by targeting mechanisms of seed coat and cotyledon adhesion with various effects on dhal quality.

BACKGROUND: Dehulling and splitting are important elements of the milling process to produce dhal from pulses. However, grain that is difficult-to-mill because of tightly adhered seed coats or cotyledons that resist separation makes it difficult to achieve high quality dhal. Milling yields are reduced, energy inputs into the milling process are increased, and the resulting dhal can be of poorer quality, chipped or abraded. RESULTS: Eight enzyme pre-treatments were chosen based on the hypothesised mechanisms of seed coat and cotyledon adhesion established previously. Using a difficult-to-mill chickpea (Cicer arietinum L.) genotype, we examined the effects of these pre-treatments, over time, on laboratory-scale milling performance and dhal quality. We pioneered a texture analyser method to measure the flex of the cotyledons and the force required to cleave the cotyledons. The enzyme-induced changes ranged from negative (tough seed coat, weight loss, deleterious colour and texture, increased visual damage to cotyledons and increased kibble loss, concave cotyledons, increased flex, and changes in taste) to positive (brittle seed coat, increased seed volume, improved dehulling efficiency and splitting yield, reduced cotyledon cleavage force, and acceptable dhal quality and taste). CONCLUSION: All pre-treatments improved milling performance compared to milling the raw seed, although there was considerable variation between them. Two pre-treatments showed no improvement in milling yields compared to the water control, and several pre-treatments resulted in unacceptable qualities. Three pre-treatments, endo-polygalacturonanase, α-galactosidase and cellulase, show potential for commercial milling applications and could assist pulse millers globally to achieve high quality dhal at the same time as minimising milling effort. © 2021 Society of Chemical Industry.

Genetic and environmental factors contribute to variation in cell wall composition in mature desi chickpea (Cicer arietinum L.) cotyledons.

Chickpea (Cicer arietinum L.) is an important nutritionally rich legume crop that is consumed worldwide. Prior to cooking, desi chickpea seeds are most often dehulled and cleaved to release the split cotyledons, referred to as dhal. Compositional variation between desi genotypes has a significant impact on nutritional quality and downstream processing, and this has been investigated mainly in terms of starch and protein content. Studies in pulses such as bean and lupin have also implicated cell wall polysaccharides in cooking time variation, but the underlying relationship between desi chickpea cotyledon composition and cooking performance remains unclear. Here, we utilized a variety of chemical and immunohistological assays to examine details of polysaccharide composition, structure, abundance, and location within the desi chickpea cotyledon. Pectic polysaccharides were the most abundant cell wall components, and differences in monosaccharide and glycosidic linkage content suggest both environmental and genetic factors contribute to cotyledon composition. Genotype-specific differences were identified in arabinan structure, pectin methylesterification, and calcium-mediated pectin dimerization. These differences were replicated in distinct field sites and suggest a potentially important role for cell wall polysaccharides and their underlying regulatory machinery in the control of cooking time in chickpea.

Near-isogenic lines of desi chickpea (Cicer arietinum L.) that differ in milling ease: differences in chemical composition.

Milling performance is an important attribute for desi chickpea and other pulses, as varieties that are more difficult-to-mill lead to processing yield loss and damage to the resulting split cotyledons (dhal) such as chipping and abrasion which are unattractive to the consumer. Poor milling performance leads to poor dhal quality and therefore lower prices and profitability along the pulse value chain. The Pulse Breeding Australia Chickpea Program identified near-isogenic desi lines that differed in seed shape and milling yields, however it was unknown whether this was due simply to a difference in physical forces on the seed during milling, mediated by seed shape, or whether there were underlying differences in chemical composition that could explain these differences. The two isolines differed in the composition of their seed coat, cotyledons and adjoining surfaces. Some of these differences were in agreement with previous research on composition of easy- and difficult-to-mill samples. These differences suggest that biochemical adhesive or cohesive mechanisms at the interface of seed tissues involve pectic polysaccharides and lignin-mediated binding.

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part II: protein, lipid and mineral composition.

BACKGROUND: Part I introduced the concept of easy- and difficult-to-mill chickpea genotypes, the broad chemical composition of their seed fractions and proposed mechanistic explanations for physical differences consistent with observed variation in milling ease. Part II continues this research by delving deeper into the amino acid, fatty acid and mineral components. RESULTS: No association between fatty acid composition and ease of milling was observed. However, particular amino acids and mineral elements were identified that further support roles of lectins, pectins and mineral-facilitated binding in the adhesion of chickpea seed coat and cotyledons. CONCLUSION: These differences suggest underlying mechanisms that could be exploited by breeding programmes to improve milling performance. This study shows that the content and composition of amino acids, fatty acids and minerals within different chickpea tissues vary with seed type (desi and kabuli) and within desi genotypes in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour.

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part I: broad chemical composition.

BACKGROUND: Ease of milling is an important quality trait for chickpeas (Cicer arietinum L.) and involves two separate processes: removal of the seed coat and splitting of cotyledons. Four chickpea genotypes (two desi types, one kabuli type and one interspecific hybrid with 'wild' C. echinospermum parentage) of differing ease of milling were examined to identify associated seed composition differences in the seed coat, cotyledons and their junctions (abaxial and adaxial). RESULTS: Several components in different fractions were associated with ease of milling chickpea seeds: primarily soluble and insoluble non-starch polysaccharides (including pectins) and protein at the seed coat and cotyledon junctions, and the lignin content of the seed coat. CONCLUSION: This study shows that the chemical composition of chickpea does vary with seed type (desi and kabuli) and within desi genotypes in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour.

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part III: free sugar and non-starch polysaccharide composition.

BACKGROUND: Parts I and II of this series of papers identified several associations between the ease of milling and the chemical compositions of different chickpea seed fractions. Non-starch polysaccharides were implicated; hence, this study examines the free sugars and sugar residues. RESULTS: Difficult milling is associated with: (1) lower glucose and xylose residues (less cellulose and xyloglucans) and more arabinose, rhamnose and uronic acid in the seed coat, suggesting a more flexible seed coat that resists cracking and decortication; (2) a higher content of soluble and insoluble non-starch polysaccharide fractions in the cotyledon periphery, supporting a pectic polysaccharide mechanism comprising arabinogalacturonan, homogalacturonan, rhamnogalalcturonan, and glucuronan backbone structures; (3) higher glucose and mannose residues in the cotyledon periphery, supporting a lectin-mediated mechanism of adhesion; and (4) higher arabinose and glucose residues in the cotyledon periphery, supporting a mechanism involving arabinogalactan-proteins. CONCLUSION: This series has shown that the chemical composition of chickpea does vary in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour. Seed coat strength and flexibility, pectic polysaccharide binding, lectins and arabinogalactan-proteins have been implicated. Increased understanding in these mechanisms will allow breeding programmes to optimise milling performance in new cultivars.

Effects of roasting on phenolic composition and in vitro antioxidant capacity of Australian grown faba beans (Vicia faba L.).

Faba bean phenolic compounds encompassed phenolic acids, flavonols, proanthocyanidins and anthocyanins. Roasting faba beans for 120 min decreased the total phenolic, flavonoid and proanthocyanidin contents by 42, 42 and 30%, respectively. Roasting beans for 120 min decreased the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, total equivalent antioxidant capacity and ferric reducing antioxidant power by 48, 15 and 8%, respectively. High performance liquid chromatography-post column derivatisation revealed the generation of new phenolic compounds as a result of roasting. Antioxidant mechanism of bean less-polar phenolic compounds was largely based on free radical scavenging activity. The bean phenolic compounds with reducing capability were heat stable. Roasted faba bean extracts (70% acetone, v/v) were fractionated into relatively polar and non-polar fractions; the latter contributed the majority of the antioxidant capacity. The extracts from beans with different seed coat colours differed in their phenolic compositions, which suggest different levels of potential benefits to health. Although roasting initially lowers the bean antioxidant capacity, prolonged roasting at 150 °C for 60 min and longer causes generation of new phenolic compounds and an increased antioxidant capacity. The findings encourage a wider ultilisation of faba beans for human foods particularly in baked/roasted products.

In vitro investigations of the potential health benefits of Australian-grown faba beans (Vicia faba L.): chemopreventative capacity and inhibitory effects on the angiotensin-converting enzyme, α-glucosidase and lipase.

The functional properties, including antioxidant and chemopreventative capacities as well as the inhibitory effects on angiotensin-converting enzyme (ACE), α-glucosidase and pancreatic lipase, of three Australian-grown faba bean genotypes (Nura, Rossa and TF(Ic*As)*483/13) were investigated using an array of in vitro assays. Chromatograms of on-line post column derivatisation assay coupled with HPLC revealed the existence of active phenolics (hump) in the coloured genotypes, which was lacking in the white-coloured breeding line, TF(Ic*As)*483/13. Roasting reduced the phenolic content, and diminished antioxidant activity by 10-40 % as measured by the reagent-based assays (diphenylpicrylhydrazyl, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) and oxygen radical absorbance capacity) in all genotypes. Cell culture-based antioxidant activity assay (cellular antioxidant activity) showed an increase of activity in the coloured genotypes after roasting. Faba bean extracts demonstrated cellular protection ability against H2O2-induced DNA damage (assessed using RAW264.7 cells), and inhibited the proliferation of all human cancer cell lines (BL13, AGS, Hep G2 and HT-29) evaluated. However, the effect of faba bean extracts on the non-transformed human cells (CCD-18Co) was negligible. Flow cytometric analyses showed that faba bean extracts successfully induced apoptosis of HL-60 (acute promyelocytic leukaemia) cells. The faba bean extracts also exhibited ACE, α-glucosidase and pancreatic lipase inhibitory activities. Overall, extracts from Nura (buff-coloured) and Rossa (red-coloured) were comparable, while TF(Ic*As)*483/13 (white-coloured) contained the lowest phenolic content and exhibited the least antioxidant and enzyme inhibition activities. These results are important to promote the utilisation of faba beans in human diets for various health benefits.

Effects of soaking, boiling and autoclaving on the phenolic contents and antioxidant activities of faba beans (Vicia faba L.) differing in seed coat colours.

The Australian grown faba beans of different seed coat colours were either soaked, boiled or autoclaved, and analysed for phenolic contents and antioxidant activity using an array of reagent-based assays. Soaking, boiling and autoclaving were shown to lower the level of active compounds in faba beans. A significant amount of active compounds was leached to the soaking and cooking medium. Boiling was a better method in retaining active compounds in beans than autoclaving. The boiled beans had more active compounds than those of resulting cooking broths, which was the opposite observation when autoclaving. The buff-genotypes had a similar level of active compounds to red- and green-genotypes. The high performance liquid chromatography-post column derivatisation (HPLC-PCD) system detected a dense collection of high antioxidant HPLC peaks ('humps') in extracts of raw, soaked and boiled beans. The present findings encouraged consumption of faba beans together with cooking broth for the maximum potential health benefits.