No tillage affects the phosphorus status,isotopic composition and crop yield of Phaseolus vulgaris in a rain‐fed farming system

BACKGROUND: Conservation tillage promotes the accretion of soil organic matter and often leads to improved soil fertility and moisture availability. However, few studies have looked at the physiological response of crop plants to different tillage practices. It was therefore hypothesised that measuring the nutrient concentrations and stable isotope composition (δ¹³C, δ¹⁸O, δ¹⁵N) of shoots could help evaluate the physiological response of common bean (Phaseolus vulgaris L.) to different tillage treatments (no tillage (NT) and mouldboard ploughing (MP)) in a rain‐fed farming system in northern Mexico. RESULTS: NT significantly enhanced shoot phosphorus concentration in bean plants. Tillage exerted a negative effect on the extent of root colonisation (%) by arbuscular mycorrhizal fungi (AMF). Lower shoot δ¹⁸O but unchanged δ¹³C values in plants from the NT system suggest enhanced stomatal conductance but also enhanced photosynthetic rate, which overall resulted in unchanged water use efficiency. Bean plants in the NT system showed lower shoot δ¹⁵N values, which suggests that a larger proportion of total plant nitrogen was obtained through atmospheric nitrogen fixation in this treatment. CONCLUSION: Greater diversity of AMF soil communities and heavier colonisation of roots by AMF in the NT compared with the MP system appeared to contribute to improved crop nutrition, water relations and yield in this rain‐fed agroecosystem. Copyright © 2010 Society of Chemical Industry     

Formation of γ‐gluconolactone in a wine‐like model system

By experiments performed in wine‐like and must‐like model solutions, we have shown that intramolecular gluconic acid esterification leads to the formation of not only the well‐known δ‐gluconolactone (glucono‐5‐lactone) but also γ‐gluconolactone (glucono‐4‐lactone). To our knowledge, the presence of the latter is reported for the first time under conditions similar to those in grape musts and wines. Equilibrium between these lactones and gluconic acid was reached within 24 h of preparing a solution of gluconic acid, and in both wine‐like and must‐like model solutions the concentrations of γ‐ and δ‐gluconolactone represented the same proportions of the acid, ie about 60 and 40 mg g^?1 gluconic acid respectively. Owing to their chemical structures, not only δ‐gluconolactone but also γ‐gluconolactone could certainly contribute to sulphur dioxide binding in musts and wines containing gluconic acid. © 2002 Society of Chemical Industry     

The role of ALA-S and ALA-D in regulating porphyrin biosynthesis in a normal and a HEM R+ mutant strain of Saccharomyces cerevisiae

Catabolite repression and derepression on δ-aminolevulinate synthase (ALA-S) and δ-aminolevulinate dehydratase (ALA-D) in a normal yeast strain, D27, and its derived D27/C6 (HEM R⁺) were investigated. ALA-S and ALA-D activities and intracellular ALA (I-ALA) at different physiological states of the cells were measured. In YPD medium, under conditions of repression and when glucose was exhausted, both strains behaved identically as if the mutation was not expressed. In YPEt medium, however, both ALA-S and ALA-D activities were higher than in YPD, but the I-ALA content and the enzymic activity profiles shown by the two strains were quite different. It appears, therefore, that the mutation causes a deregulation of ALA-S, so that its activity is kept at a high level throughout the cell cycle. This would explain the increased levels of cytochromes present in the mutant. This mutation may affect some regulatory aspect of ALA formation and renders an ALA-S of high activity; moreover, this enzyme species seems to be more stable than in the normal strain.     

Influence of Heat Pretreatments of Oat Grain on the Viscosity of Flour Slurries

Heat treatment of oat grain had significant effects on the viscosity of flour slurries. Steamed oats produced highly viscous flour slurries, whose viscosity increased with time, whereas the viscosity of flour slurries produced from raw or roasted (104°C for 120 min) oats was much lower and degraded rapidly. Slurry viscosity was correlated with (1→3), (1→4)-β-D -glucan concentration in the flours and treatment of slurries with lichinase lowered viscosity significantly. Effects of steaming were partially reversed by roasting treatment and vice versa. Mixtures of equal amounts of raw and steamed flour resulted in slurries more viscous than either flour alone, but that viscosity degraded after 3 h to less than the mean viscosity of the steamed and raw controls. Water-soluble extracts from steamed flour had about twice the viscosity of raw or roasted flour extracts, but contained only 80% of the (1→3), (1→4)-β-D -glucan present in those extracts. Molecular weight analysis of soluble carbohydrates from raw, roasted and steamed soluble extracts indicated the molecular weight of β-glucans in these extracts was similar. However, if extracts were made from slurries that had incubated for 3 h, extensive degradation of β-glucans was evident in raw and roasted samples. It is likely that enzymic degradation of (1→3),(1→4)-β-D -glucans is responsible for much of the decreased raw and roasted flour slurry viscosity over extended time periods, but different heat treatments appear to also affect (1→3), (1→4)-β-D -glucan polymer interaction. © 1997 SCI.     

Characterisation of lactose crystallised in acidified aqueous ethanol

Three types of crystalline lactose hydrates having melting points (MP) of 218°C, 195°C and 169°C were crystallised from 680 ml litre^?1 ethanol solution with a lactose to solvent ratio of 1:14-1:24 at three pH values, ～ 4, 2.5 and 1.3. The three types were characterised on the basis of their optical rotation, phase transition and X-ray powder diffraction pattern. The samples with high and low MP were α-lactose hydrates whereas that with the intermediate MP (195°C) was a hydrated lactose containing both α- and β-forms.     

ENZYMIC QUANTIFICATION OF (1→3) (1→4)-β-D-GLUCAN IN BARLEY AND MALT

A simple and quantitative method for the determination of (1→3) (1→4)-β-D-glucan in barley flour and malt is described. The method allows direct analysis of β-glucan in flour and malt slurries. Mixed-linkage β-glucan is specifically depolymerized with a highly purified (1→3) (1→4)-β-D-glucanase (lichenase), from Bacillus subtilis, to tri-, tetra- and higher degree of polymerization (d.p.) oligosaccharides. These oligosaccharides are then specifically and quantitatively hydrolysed to glucose using purified β-D-glucosidase. The glucose is then specifically determined using glucose oxidase/peroxidase reagent. Since barley flours contain only low levels of glucose, and maltosaccharides do not interfere with the assay, removal of low d.p. sugars is not necessary. Blank values are determined for each sample allowing the direct measurement of β-glucan in maltsamples.α-Amylasedoes not interfere with the assay. The method issuitable for the routineanalysis of β-glucan in barley samples derived from breeding programs; 50 samples can be analysed by a single operator in a day. Evaluation of the technique on different days has indicated a mean standard error of 0–1 for barley flour samples containing 3–8 and 4–6% (w/w) β-glucan content.     

COMPARATIVE STUDIES OF THE β-D-GLUCAN RELEASED INTO SORGHUM AND BARLEY WORTS

Worts prepared from two cultivars of Nigerian grown sorghum six day melts — LI87 end SK5912 had β-D-glucan levels off five to seven times more than that of proctor barley. In contrast to barley, malting of the sorghums results in the release off more β-D-glucan into wort. Apparently, this is due to increasing levels of β-glucan solubilase and (1→3)-β-glucanase during malting with no significant (1→3, 1→4)-β-glucanase activity.     

THE INSTITUTE OF BREWING ANALYSIS COMMITTEE THE DETERMINATION OF ENDO-β-GLUCANASE ACTIVITY IN MALT

Various substrates for the determination of malt endo-β-glucanase activity have been tested in conjunction with the β-glucanase procedure advocated by Bourne and Pierce. It is recommended that one universal standard substrate should be used and that each determination should be measured against a standard check malt. Furthermore, great care must be exercised in the dissolution of the substrate to obtain consistent results.     

CHANGES IN BARLEY ENDOSPERMS DURING EARLY STAGES OF GERMINATION*

Kernels of 4 barley cultivars were germinated at 18°C and samples were removed for analysis at short time intervals for the first 30 h and at longer intervals during the ensuing 90 h. α-Amylase, (1 → 3) (1 → 4)-β-glucanase and (1 → 3) β-glucanase activities were measured in each sample. Analysis of kernel sections stained with Calcofluor showed that hydrolysis of β-glucan in the crushed cell layer commenced 6–9 h after the initiation of germination. Hydrolysis proceeded from the ventral edge to the dorsal edge of the kernel. Starch granule hydrolysis followed a similar pattern in the endosperm region adjacent to the crushed cell layer, but starch hydrolysis was always preceded by β-glucan hydrolysis.     

AN APPROACH TO THE IDENTIFICATION OF A ß-GLUCAN SOLUBILASE FROM BARLEY1

A ß-glucan solubilase activity was demonstrated in barley extract. This enzyme catalyzed the dissolution of barley ß-glucan by releasing a product having a narrow molecular weight distribution and a molecular weight of about 20,000. The enzyme was partially purified by ion exchange chromatography on DEAE-cellulose and gel permeation chromatography on Bio-Gel P-100. Although carboxypeptidase activity was present in the crude extract of barley flour the partially purified ß-glucan solubilase did not hydrolyse N-CBZ-Phe-ala. Examination of extracts from different barley tissues indicated that the ß-glucan solubilase activity was associated with the husks only; a large portion of the activity was extractable from whole barley kernels. About 85% of the enzyme activity in crude extracts from barley flour was retained after 40 min at 62°C. However, the enzyme was much more heat-labile in extracts of whole barley kernels. The pH of maximal activity was found to be about pH 5.7 and results from column chromatography suggested that the enzyme had a low pl value and a MW between 5 × 10⁴ and 6 × 10⁴.     

THE APPLICATION OF SALT FORMATION IN THE CHEMISTRY AND TECHNOLOGY OF HOP RESINS

In natural product chemistry the satisfactory isolation of chemical compounds of scientific and/or commercial significance normally follows a pattern of fractionation which involves the utilization of a range of physical and chemical properties of the compounds present in the mixture. Vapour pressure and solubility are the physical properties traditionally made use of in the initial stages of fractionation. Chemical properties normally enable somewhat more specific fractionation to be achieved. One chemical property that is widely used is the ability of a large number of compounds to form salts. In the case of the hop resins it is this specific chemical property which has played the most significant role in their isolation, characterization and commercial utilization. The historical development of this aspect of hop chemistry has been reviewed.     

Gamma/alpha‐zein hydrolysates as oral delivery vehicles: Enhanced physicochemical stability and in vitro bioaccessibility of curcumin

In this study, we aimed to evaluate the potential of the γ‐zein hydrolysate (γ‐ZH) as a new delivery system for hydrophobic bioactive molecules such as curcumin (Cur). γ‐ZH‐Cur complexes (γ‐ZH‐Cur) with a particle size of 50–60 nm were successfully formed via the hydrogen bonding and hydrophobic interactions. Compared to α‐ZH, the complexation of Cur in γ‐ZH showed a better improvement in the water solubility and physicochemical stability of Cur and showed sustained release and greater potential for absorption of Cur. The differences in the loading capability, the physicochemical storage and the bioaccessibility of Cur between γ‐ZH and α‐ZH may be associated with the differences in the amino acid composition and typical sequence of peptides. These results indicate that γ‐ZH has a better ability to serve as a novel delivery for the hydrophobic bioactive molecules compared to α‐ZH.     

IONIZATION BEHAVIOUR OF HOP COMPOUNDS AND HOP-DERIVED COMPOUNDS

The ionization behaviour of colupulone, (?)-humulone, trans-isohumulone and trans-humulinic acid was investigated using a number of techniques. When a potentiometric technique was employed, precise estimates of pKa could not be obtained for any of the compounds, since a drift in the observed pKa values occurred during the titrations. Approximate, or equilibrium pKa values for the most acidic function of each of the compounds were: colupulone, 6.1; (?)-humulone, 5.0; trans-isohumulone, 3.1; trans-humulinic acid, 2.7. Evidence from spectroscopic measurements (UV, ¹H-NMR, ¹³C-NMR), conductiometric experiments and measurements of the pH dependency of solvent partition behaviour and aqueous solubility of the compounds is consistent with the possibility that, in aqueous solution, compounds such as trans-isohumulone are present as covalent hydrates. It is likely that reversible hydration occurs at one or more carbonyl groups. As a consequence of this behaviour, no single pKa value is sufficient to describe the behaviour of any one hop compound or hop-derived compound. The potential significance of this phenomenon is discussed.     

IMPROVED EXTRACTION AND ASSAY OF β-AMYLASE BROM BARLEY AND MALT

β-Amylase was extracted from barley or malt using four physical techniques to break up grists which had been prepared using a Moulinex coffee grinder. Grinding with a Polytron homogeniser apparently completely disrupted all cells, as determined by transmission electron microscopy, and increased the efficiency of extraction of β-amylase from barley by more than 30%. The other treatments tested were without value . The β-amylase activity in extracts of barley or malt was assayed by measuring the production of reducing sugars from reduced soluble starch, using a PAHBAH reagent. α-Amylase, which interferes with the quantitation of β-amylase in extracts of malt, was not totally inactivated by the chelating buffer used for enzyme extraction or by several other chelating agents. α-Amylase activity was quantified specifically using Phadebas. Using purified α-amylase a calibration was developed which related activity, as determined using Phadebas, to reducing power units. Thus the α-amylase activity present in an extract containing β-amylase could be determined using Phadebas and the reducing power equivalent activity subtracted from the total “apparent” activity to give the actual β-amylase activity. α-Glucosidase and limit dextrinase activities are believed to be too low to have a significant effect on the apparent β-amylase . The soluble and bound β-amylase activities were measured in samples taken from micromalting barley (Alexis). Dry weight losses increased to over 10% after 8 days germination. Antibiotics, applied during steeping, were used to control microbes in one experiment. However, their use checked germination and reduced malting losses to 8.4% in 8 days germination. The soluble enzyme present in extracts from steeped barley and early stages of germination was activated (20–40%) by additions of the reducing agent DTT .     

Characterization of lupin major allergens (Lupinus albus L.)

White lupin is considered to be a rich source of protein with a notable content of lysine and is being increasingly used in bakery, confectionery, snacks and pastry products due to its multifunctional properties, in addition to its potential hypocholesterolemic and hypoglycemic properties. However, lupin seed flour has been reported as a causative agent of allergic reactions, especially in patients with allergy to peanut since the risk of immunological cross‐reactivity between lupin and peanut is higher than with other legumes. Previously, we had identified two proteins as major lupin allergens (34.5 and 20 kDa) as determined by IgE immunoblotting using sera of 23 patients with lupin‐specific IgE. The aim of this study was to purify and characterize the two major lupin allergens. The results using in vitro IgE‐binding studies and MS analysis have shown that the 34.5 kDa allergen (Lup‐1) is a conglutin β (vicilin‐like protein) while the 20 kDa allergen (Lup‐2) corresponds to the conglutin α fraction (legumin‐like protein). The high level of amino acid sequence homology of Lup‐1 and Lup‐2 with the major allergens of some legumes explains the IgE cross‐reactivity and clinical cross‐reactivity of lupin and other legumes.     

Hydrolysis of β-glucosyl ester linkage of p-hydroxybenzoyl β- -glucose, a chemically synthesized glucoside, by β-glucosidases

To investigate the hydrolysis of glucosyl esters by β-glucosidase, p-hydroxybenzoyl β- -glucose (pHBG) was chemically synthesized. The hydrolytic activity of some β-glucosidases for pHBG was compared to that for p-nitrophenyl β-glucoside (pNPG). The Clavibacter michiganense and Flavobacterium johnsonae enzymes could hydrolyze pHBG and steviol glycosides which are natural glucosyl esters. The commercial β-glucosidase originating from Caldocellum saccharolyticum also hydrolyzes pHBG despite having no activity for steviol glycosides. The β-glucosidase from Aspergillus niger cleaved the glucosyl ester linkage much more weakly than the glucosidic linkage. The pH-activity profile for the hydrolysis of pHBG was similar to that of pNPG by the C. saccharolyticum β-glucosidase. The similar profiles for these substrates suggested that the active site for the glucosyl ester chemically resembles that for glucoside with respect to catalysis. Kinetic analysis of the C. saccharolyticum β-glucosidase for mixed substrates of pHBG and pNPG showed that the hydrolysis of pHBG competed with that of pNPG. This result indicated that there is only one active site for both the glucosyl ester and glucoside. Mass spectroscopic analysis of the hydrolysates of pHBG in H₂¹⁸O suggested that β-glucosidase hydrolyzes glucosyl esters between the anomeric carbon of glucose and the carbonyl oxygen, not between the carbonyl carbon and the carbonyl oxygen.     

β-GLUCANASE: THE SUCCESSFUL APPLICATION OF GENETIC ENGINEERING

The now well established principles of genetic engineering are described in relation to the solution of problems associated with β-glucans in beer. The application of these techniques has enabled the isolation of a Bacillus subtilis endo-1, 3–1, 4-β-D-glucanase gene which expresses a biologically active enzyme in yeast.^15,16 Although this enzyme is capable of hydrolysing beer β-glucans during fermentation, thereby enhancing beer filtration, insufficient β-glucanase is produced in yeast to enable successful commercial implementation. The requirements for the efficient production of β-glucanase in genetically manipulated brewing yeast are described.     

THE DETERMINATION OF α- AND β-ACIDS IN HOPS AND HOP PRODUCTS USING HPLC

A rapid reversed phase HPLC method for the analysis for α- and β-acids in hops and hop products is described and has been evaluated. The method uses citric acid in the eluent as a complexing agent to overcome the irreversible adsorption effects shown by some columns, thus allowing optimum eluent pH to be selected. The precision of the method for analysis of hop extract has been determined giving relative standard deviations of 1·0% and 2·1% for α- and β-acids respectively. General agreement with results obtained using a polarimetric α-acids analysis method for hop extracts and hops has been demonstrated.