The effect of gluten on the retrogradation of wheat starch

The retrogradation of amylopectin in a wheat starch and a wheat starch/gluten (10:1) blend prepared by extrusion and containing 34% water (wet weight basis) was studied using X-ray diffraction, differential scanning calorimetry and NMR relaxometry during storage at constant water content and temperature (25 °C). For both samples, amylopectin ‘fully’ retrograded after 2–3 days storage, i.e. the different parameters monitored with time to follow the retrogradation had reached their maximum value, and crystallised predominantly into the A polymorph. Under the experimental conditions used, there was no evidence of any significant effects of the presence of gluten on the kinetics, extent or polymorphism of amylopectin retrogradation.     

Effect of particle size on kinetics of starch digestion in milled barley and sorghum grains by porcine alpha-amylase

The influence of milled grain particle size on the kinetics of enzymatic starch digestion was examined. Two types of cereals (barley and sorghum) were ground, and the resulting grounds separated by size using sieving, with sizes ranging from 0.1 to 3 mm. In vitro enzymatic digestion was performed, using pancreatic alpha-amylase, amyloglucosidase and protease, to determine fractional-digestion rates over 24 h. The resulting glucose production rate data were well fitted by simple first-order kinetics. For each sieve screen size, the digestion rate of barley was always higher than that of sorghum. The rate coefficients for digestion showed a decrease with increasing size, and could be well fitted by an inverse square relationship. This is consistent with the supposition that starch digestion in these systems is controlled by diffusion of enzyme through the grain fragment. Apparent diffusion coefficients of alpha-amylase obtained by fitting the size dependence were 0.76 (sorghum) and 1.7 (barley) × 10⁻⁷ cm² s⁻¹, 9 (sorghum) and 4 (barley) times slower than predicted for a molecule of the size of alpha-amylase in water.     

Identification of quantitative trait loci for β-glucan concentration in barley grain

The amount of (1 → 3),(1 → 4)-β-d glucan (β-glucan) accumulated in cell walls of barley (Hordeum vulgare L.) kernel is an important determinant for grain end-use. Grain β-glucan concentration is affected by environmental and genetic factors and usually varies from 3 to 6%. In this study, we have analyzed the β-glucan trait in a doubled-haploid (DH) population of 170 lines grown in three separate field trials. Most of the DH lines showed β-glucan values that ranged from that of the low β-glucan parent (cultivar CDC Bold; 3.3%) to that of the high β-glucan parent (breeding line TR251; 5.4%). Eighty-eight lines of the DH population were genotyped using simple sequence repeat (SSR), amplified fragment length polymorphism (AFLP) and diversity array technology (DArT) markers, which were subsequently integrated into a barley genetic map spanning 1059 cM. Interval mapping and multiple-QTL-mapping (MQM) of quantitative trait loci (QTL) from the three trials indicated seven genomic regions associated with low grain β-glucan concentration. For all putative QTLs, the low β-glucan concentration was contributed by alleles from CDC Bold except for two loci on chromosomes 5H that were derived from TR251. A major QTL located to the centromere region of chromosome 7H was identified by both mapping methods for all three trials. The 7H QTL explained up to 39% of the β-glucan concentration and genetic markers associated with the locus may be used to aid selection of high and low β-glucan barley lines.     

The identification of a barley haze active protein that influences beer haze stability: The genetic basis of a barley malt haze active protein

In bright beers, the formation of haze is a serious quality problem, which places limitations on the storage life of the product. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) immunoblot analysis using an antiserum that was raised against a silica eluate (SE) protein fraction (obtained from silica gel, used for the colloidal stabilisation of beer), detected a range of protein bands in barley, malt, beer and haze. A polymorphism was observed in which some barley varieties contained a molecular weight (MW) 12,000 band (SE +ve) while in other varieties this band was absent (SE −ve). A survey of 219 Australian and international barley varieties, including a comprehensive selection of current and past malting varieties, identified 181 varieties as SE +ve, and 38 varieties as SE −ve. Previous pilot brewing trials demonstrated that SE −ve varieties are desirable as the beer brewed from the malt of these varieties formed less haze after accelerated ageing than beers brewed using SE +ve malt varieties. The genetic basis for the absence of the SE protein was conferred by a recessive allele at a single locus. Interval mapping analysis showed that the MW 12,000 band mapped to the short arm of chromosome 3H.     

Relationship between the glass transition temperature and the melt flow behavior for gluten, casein and soya

The effects of moisture content (25–45% wwb) and temperature (75–120 °C) on the viscosity of gluten, soya and rennet casein systems was studied using a capillary rheometer. An attempt was made to relate the viscosities to the glass transition temperature measured by differential scanning calorimetry, dynamic mechanical thermal analysis and the phase transition analyzer. The temperature where the material flowed was also determined by the latter technique. All three-protein systems showed shear and extension thinning. Over the shear rate range investigated (1–10³ s⁻¹), gluten had a substantially lower viscosity than the other two proteins, although the difference was less pronounced at the highest temperature studied. This low viscosity is reflected by lower values of the glass transition temperature, the melt flow temperature and the dynamic moduli E′ and E″ in the rubbery state. The results are discussed in terms of the structure and heat induced changes for the three proteins and their relevance to food processing considered.     

Postharvest decay control and quality retention in litchi (cv. McLean's Red) by combined application of modified atmosphere packaging and antimicrobial agents

The effects of modified atmosphere packaging (MAP) [bioriented polypropylene (BOPP-1 or BOPP-2)] in combination with antimicrobial agents Bacillus subtilis, 10⁷ colony-forming units (cfu) ml⁻¹; ethylenediamine tetraacetic acid, calcium disodium salt hydrate (EDTA) (0.1%); or 4-hexylresorcinol (4-HR) (0.15%) on postharvest decay control and quality retention of litchi cv. McLean's Red were assessed as possible replacements for commercial SO₂ fumigation. Fruits dipped in B. subtilis, EDTA or 4-HR (5 min) separately, blow dried (25 °C, 3 min), packed in BOPP-1, held for 18 d at 2 °C, 95% RH, and 2 d at 14 °C, 75% RH were significantly less decayed. The antagonist–BOPP-1 combination also promoted the best bacterial survival during storage. B. subtilis was observed to survive effectively in BOPP-1 (16% O₂, 6% CO₂; 90% RH), but its survival was adversely affected in BOPP-2 (5% O₂, 8% CO₂; 93% RH). Alternaria alternata and Cladosporium spp. were the major decay-causing fungi in BOPP-1 treatments, and Candida, Cryptococcus and Zygosaccharomyces were the predominant yeasts in BOPP-2 treatments. Combination treatments EDTA, 4-HR or B. subtilis in BOPP-1 inhibited polyphenol oxidase (PPO) and significantly reduced pericarp browning and severity. Although the combination treatments EDTA, 4-HR or B. subtilis in BOPP were equally effective in controlling decay and browning, the EDTA and 4-HR affected the natural pinkish-red colour of the pericarp by showing higher h° values (orange–pink). Among the combination treatments, B. subtilis+BOPP-1 had the best potential to control decay, retain the colour and the overall litchi fruit quality during a marketing chain of 20 d.     

The identification of a barley haze active protein that influences beer haze stability: Cloning and characterisation of the barley SE protein as a barley trypsin inhibitor of the chloroform/methanol type

Previous pilot brewing trials have demonstrated that in the absence of a molecular weight (MW) 12,000 (barley silica eluate (SE) protein), the beer brewed from the malt of these SE −ve varieties formed less haze after accelerated ageing than beers brewed using SE +ve malt varieties. The previously described SE protein was characterised using comparative two-dimensional (2-D) gel electrophoresis immunoblots of barley seed extracts from both SE +ve and SE −ve varieties. The SE protein spot identified was excised and its partial sequence determined, after in-gel cleavage using trypsin and separation of the resulting fragments by reversed-phase high performance liquid chromatography (HPLC). N-terminal sequence analysis of the tryptic peptides from SE +ve and SE −ve varieties identified the SE protein as the barley trypsin inhibitor-CMe precursor (BTI-CMe). The mature BTI-CMe protein is 13.3 kDa and its functional gene is located on chromosome 3H. Cloning of the BTI-CMe protein demonstrated that both SE −ve and SE +ve barley varieties contain a BTI-CMe protein family member that is similar but has a consistently different sequence, primarily in the last 30 amino acid residues of their C-termini.     

Salt effect on yield and composition of shoot essential oil and trichome morphology and density on leaves of Mentha pulegium

The aim of the present work was to study salt effect on the yield and composition of shoot essential oil (EO) and the structures responsible for its biosynthesis in Mentha pulegium L. Shoot EO was extracted by hydrodistillation and composition was determined by GC–MS method. Apical and basal leaves were taken for microscopy analyses; small fresh samples were observed directly without fixation or metallisation with environmental scanning electron microscope (ESEM) and stereomicroscope (SM). Fresh separate epidermis was used for light microscopy (LM). Salt stress enhanced EO yield by about 2.75 times and affected the percentage of menthone, which is the major compound (51%), increasing that of pulegone. Menthone, pulegone, and neomenthol constituting the monoterpene class were found to be the principal components. The anatomical study showed three types of trichomes: (i) non-glandular, multicellular, simple hairs; (ii) small, capitate glandular trichomes; (iii) and peltate glandular trichomes. In control plants, the density and size of trichomes varied with leaf side (abaxial or adaxial) and developmental stage. Salt stress results in significant modifications affecting trichome distribution and size on both sides.     

Arabinoxylan and hydroxycinnamate content of wheat bran in relation to endoxylanase susceptibility

Hydroxycinnamate and protein contents and monosaccharide composition were determined for 11 industrial wheat (Triticum aestivum) brans and related to the susceptibility of their arabinoxylans (AX) to enzymatic degradation. There was significant variation in carbohydrate, A/X ratio, protein, hydroxycinnamic acid and diferulic acid (DiFA) content among the wheat brans. In addition, a strong correlation was found between AX and ferulic acid contents. Brans were extracted with water followed by a dry-thermal treatment to remove 90% starch and to inactivate endogenous enzymes and proteinaceous inhibitors. Treated brans were compared with respect to AX degradation by a family 11 xylanase. Digestion with xylanase had a strong impact on the chemical composition of the residual bran. The A/X ratio changed from 0.60 to 1.07. The solubilised AX had an A/X ratio of 0.32. The A/X ratio of enzyme-depleted bran was negatively correlated with the loss of AX. Total DiFA in destarched brans was negatively correlated with the amount of soluble AX. These relationships indicate that structural features of AX and the extent of its cross-linking in the cell walls of the bran tissues influence its susceptibility to xylanase treatment. Thus, the amount of enzyme-solubilised AX was not directly related to the content of AX in the destarched bran. However, brans differed in their susceptibility to xylanase attack.     

Lipid oxidation and amylopectin molecular weight changes occurring during storage of extruded starch samples

Using thermomechanical extrusion, waxy maize starch and 4% (w/w) lipid were formed into strips. The lipids used were free fatty acids (mostly linoleic) and antioxidant stripped sunflower oil (either previously oxidised or fresh, with and without copper ions). By altering their water content it was possible to store, at the same temperature, sample strips in the glassy and rubbery states. Lipid oxidation was monitored by determining hexanal in the headspace above all stored samples. The molecular weights of the amylopectin in the samples of extruded products (dissolved in dimethylsulfoxide, precipitated and redissolved using a pressure cell) were determined by asymmetric flow-field-flow fractionation, analytical ultracentrifugation and light scattering. The initial rate of hexanal generation was higher in samples stored in the glassy state compared with those in the rubbery state. Waxy maize and extruded samples, at the start of the storage period and after 42 days, were used to establish the molecular weight of the amylopectin. A significant fall (a decrease of 40%) in molecular weight was found on storage of samples containing sunflower oil and copper ions and those containing free fatty acids, irrespective of the method used for molecular weight determination.     

Molecular Basis of the Gelatinisation and Swelling Characteristics of Waxy Rice Starches Grown in the Same Location During the Same Season

Six waxy rice samples (grown at the same site during the same season) were investigated to identify those aspects of amylopectin structure and granule composition which differentiate the starches into high (T_p 77·8 °C on average) or low (T_p 66·2 °C on average) gelatinisation temperature. All starches contained less than 2·5% amylose with negligible lipid. Granule dimensions were similar (mean diameter 4·9–5·7 μm) as was the β-amylolysis limit (59·9–63·0%). However, the structural elements of amylopectins within the two groups varied. The low gelatinisation temperature starches contained two major chain fractions upon debranching with iso-amylase, F3 (DP 16) and F1 (DP 51) on average while the high gelatinisation temperature starches contained three fractions, F3 (DP 16), F2 (DP 19) and F1 (DP 40). The proportion of F1/(F2+F3) for the low and high gelatinisation starches representing on average 20·2 and 10·5% respectively. Hence, the higher proportion of exterior chains and especially those with DP 19 appear to confer the higher gelatinisation temperature characteristic of these starches. Debranched β-limit dextrins were prepared from these starches. Regardless of their origin, (high or low gelatinisation temperature), they contained three fractions: F_3β (DP 5), F_2β (DP 13) and F_1β (DP 19). Hence, the high versus low gelatinisation characteristic was imparted by the exterior chains. Using 13-C CP-MAS/NMR it was confirmed that the number of double helices within the starches were approximately constant although small differences in crystallinity were identified by X-ray diffraction. Retrogradation of the solubilised native high and low gelatinisation temperature amylopectin molecules confirmed that the ‘high temperature gelatinisation amylopectin’ molecules form higher dissociation temperature and enthalpy crystalline domains. This confirms that the higher proportion of short and especially relatively longer short chains promotes crystallite formation.     

Spectroscopic comparison of lignin separated by electrolysis and acid precipitation of wheat straw soda black liquor

Electrolysis experiments were carried out with wheat straw soda black liquor. Organics from black liquor were electrodeposited at the anode. These organics were studied for their FTIR and NMR spectra and compared with those separated by acid precipitation of black liquor. The electrodeposited organics were found to be chemically distinct from those obtained by the acidification of the black liquor. Notably, they differed in the characteristic features of the aromatic part from hitherto known lignin formulations as revealed by the absence of signals for quaternary aromatic carbon between δ 129 and δ155 ppm in their ¹³C NMR spectra and higher proportion of aromatic protons in their ¹H NMR spectra. The hydroxyl environment was also found to be different by the shift in OH stretching band to higher frequency. Other significant differences were noted in the FTIR and NMR spectra of the electrodeposited solids from those of acid precipitated solids.     

Purification and Characterisation of Ascorbate Oxidase from Flour and Immature Wheat Kernels

White flour from wheat was shown to contain basic-ascorbate oxidase (AOX) enzymes (pI 7·6–9·6) and acidic-AOX enzymes (pI 5·1–6·6) in a ratio of 0·4:1, based on chromatography data. Immature wheat kernels (two weeks post-anthesis) contained about 12 times more AOX activity (units/g dry weight) than flour from mature grain, and the ratio of basic- to acidic-AOX was 5:1. Acidic-AOX was purified 90-fold from flour by hydrophobic interaction, gel filtration and anion exchange chromatography. Basic-AOX was purified 20 000-fold from immature wheat by hydrophobic interaction, anion exchange, cation exchange and gel filtration chromatography in a yield of 5%. The acid-AOX had a M of 140 k, was optimally active at pH 6·3 and 40 °C, and was stable in the pH range 5–9 and at 30 °C for 0·5 h at pH 6·2. The Km values were 0·26 m for L-ascorbic acid and 0·93 m for D-iso ascorbic acid. The basic-AOX had a M of 139 k and subunit M of 72 k. The enzyme was optimally active at pH 6·2 and 50 °C, and was stable in the pH range 5–9 and at 40 °C for 0·5 h at pH 6·2. The Km values were 0·30 m for L-ascorbic acid and 0·53 m for D-iso ascorbic acid. The absorption spectrum of basic-AOX had absorption maxima at 280 nm and 607 nm of similar magnitude to those measured in AOX fromCucurbita species (squash). This indicates that wheat AOX contains protein-bound copper similar to other plant AOX.     

Weed control and sesame (Sesamum indicum L.) response to preplant incorporated herbicides and method of incorporation

Field studies were conducted at two sesame-growing regions of Texas in 2004 and 2005 to determine weed control and sesame response to four dinitroaniline herbicides and their method of incorporation. Ethalfluralin, pendimethalin (formulated as Prowl EC), and trifluralin were applied at the X, 1X, and 2X of the suggested label dose for Gossypium hirsutum L. Pendimethalin (formulated as Prowl H₂O) was applied at the X, 1X, and 1X rate. Two methods of incorporation included rolling cultivator mixing wheels and spring tooth harrow. With rolling cultivator mixing wheels, all herbicides controlled Amaranthus tuberculatus (Moq.) J.D. Sauer at least 74% regardless of dose. The X dose of ethalfluralin and pendimethalin (formulated as Prowl EC), or the X and 1X dose of pendimethalin (formulated as Prowl H₂O) controlled Brachiaria platyphylla (Griseb.) Nash no better than 73% while all other doses of the herbicides controlled B. platyphylla at least 80%. The use of mixing wheels to incorporate the herbicides resulted in better sesame stands and less stunting than the use of the spring tooth harrow; however, sesame stands were reduced as herbicide rate increased when using mixing wheels. Ethalfluralin at the 1X dose, pendimethalin (formulated as Prowl H₂O) at the X dose, and trifluralin at the X dose produced the highest sesame yield while ethalfluralin at the 2X dose produced the lowest yield.     

Effect of Chemical Treatments on Polyphenols and Malt Quality in Sorghum

Tannin-containing sorghums, Chirimaugute and DC-75, and a tannin-free sorghum, SV2, were steeped in water, HCl (0·25 ), formaldehyde (0·017 ) and NaOH (0·075 ) for 8 and 24 h. Germination was carried out for 2 and 5 d. Steeping in NaOH enhanced water uptake of the grains compared with the other treatments. All treatments reduced the polyphenol content of the raw grain. Treatment with NaOH or formaldehyde (HCHO) was more effective than water or HCl. Malt quality was measured in terms of diastatic power (DP). Potential DP, determined after the peptone extraction, indicated higher amylase activity in DC-75 malt than in Chirimaugute and SV2 malt. Available DP, determined after water extraction, was low in malt from the tannin-containing varieties that had been treated with water or HCl. Malting alone was not an effective method of reducing the enzyme inhibitory power of the sorghum tannins. Available DP was markedly improved by the NaOH and HCHO treatments of the tannin-containing varieties. It is concluded that steeping in dilute NaOH is effective in detoxifying high-tannin sorghums, reducing the steeping period and enhancing malt quality. Steeping in NaOH appears to be a safer alternative to HCHO for treatment of high-tannin sorghums in the malting industry and for other food uses.     

Assessment of antioxidant activity and rheological properties of wheat and buckwheat milling fractions

This study examined the antioxidant properties of the ethanolic extracts of wheat milling fractions (wheat flour type 500 and type 850, and bran) and their polyphenol and tocopherol content, and rheological characteristics of wheat dough supplemented with buckwheat flours (light and wholegrain). The results obtained in this study were correlated with our previously published data on wheat flour type 400, wholegrain wheat flour and buckwheat flours.Buckwheat flours exhibited significantly higher (P < 0.05) antiradical activity on hydroxyl (OH), superoxide anion (O₂⁻) and (1,1-diphenyl-2-picrylhydrazyl) DPPH radicals, antioxidant activity and reducing power than all investigated wheat milling fractions when their corresponding IC₅₀ values were compared.The rheological parameters of wheat dough supplemented with light and wholegrain buckwheat flour (0-50%) were obtained by using Mixolab. Results indicated changes in protein and starch properties of dough.The obtained results indicate the benefit of using buckwheat flours in wheat-based food products, i.e. their contribution in functional and tailor-made-food production.     

Nematicidal efficacy of isothiocyanates against root-knot nematode Meloidogyne javanica in cucumber

Isothiocyanates (ITCs), a series of new nematicides of the -NCS group, were evaluated for their efficacy against root-knot nematode Meloidogyne javanica. Of the compounds tested, AllylITC, AcITC, EtITC, BzTC, BzITC, 1-PEITC and 2-PEITC showed in vitro irreversible nematicidal activity against second-stage juveniles of M. javanica, following exposure for 72 h at concentrations as low as 5 μg mL⁻¹. When exposed to AllylITC, AcITC and EtITC at lower concentrations, motile juveniles also became irreversibly immobile in 3 days, with a LC₅₀ value at 2.76, 2.53 and 3.05 μg mL⁻¹, respectively. In the pot experiments, 1.0 ml AllylITC and 1.1 ml AcITC per kg of soil controlled M. javanica, similarly to or better than metam sodium at its recommended dose. Similar results were obtained in the field experiments using 1.0 kg AllylITC or 1.0 kg AcITC ha⁻¹. Based on the results of this study, AllylITC and AcITC have potential to be used as new bio-fumigant nematicides.