Sweet taste and solution properties of α,α-trehalose

α,α-Trehalose, a naturally occurring non-reducing disaccharide (α, D-glucopyranosyl 1–1 α-D-glucopyranoside) is investigated by solution parameters and sensorial effects. Interactions between α, α-trehalose and water give rise to hydration effects which are measurable by intrinsic viscosity [n], apparent specific volume V°₂, and NMR relaxation times. Water-solute interactions are prime determinants of taste qualities. An interpretation of sweetness intensity and persistence of α,α-trehalose is therefore offered, based on its solution properties and its effect on water structure. From the results, it appears that α,α-trehalose undergoes a better packing among water molecules than other sugars. α,α-Trehalose is found to be less sweet but more persistent than sucrose, fructose, glucose or maltose.     

SIMULTANEOUS DETERMINATION OF α, β AND ISO-α ACIDS IN HOPS AND HOP PRODUCTS

A method for the simultaneous analysis of α, β and iso-α acid in hops, hop extracts and isomerised hop extracts is described. It is based on the use of reversed phase high performance liquid chromatography and quantitative evaluation of the hop compounds is carried out with a computing integrator. The isomerisation reaction can be examined in detail, particularly in connection with the production of hop derived haze forming compounds in isomerised hop extracts used for post fermentation bittering.     

Stabilization of soybean oil body emulsions using κ, ι, λ-carrageenan at different pH values

Oil bodies, with their unique structural proteins, oleosins, are known to be useful in foods and other emulsion systems. The influence of ??, ??, and ??-carrageenans on the stability of soybean oil body emulsions at different pH values (pH 3, 4, 5 and 7) was investigated by particle electrical charge, particle size distribution, creaming stability and confocal laser scanning microscopy measurements. In acidic environment (pH 3, 4 and 5), the droplet charge of soybean oil body emulsions stabilized with carrageenan decreased with increasing carrageenan concentration for all types of carrageenan investigated, suggesting their adsorption to the oil body droplet surfaces. Extensive droplet aggregation and creaming were observed in the emulsions stabilized with ??-carrageenan at pH 3 and 5, indicating that soybean oil body droplets were bridged by carrageenan. At pH 7, there was no significant change in the droplet charge of soybean oil body emulsions stabilized with three types of carrageenan, but the emulsions stabilized with ??-carrageenan were more stable to creaming due to depletion flocculation than the emulsions stabilized with ?? or ??-carrageenan after seven days storage. The probable reason was that ??-carrageenan, which had the most densely charged helical structure, was most effective at creating highly charged interfacial membranes, thus reducing the depletion flocculation to occur.     

Enzymatic Characterisation of Novamyl®, a Thermostable α-Amylase

The thermostable -amylase Novamyl^® is used in the baking industry as an antistaling agent due to its ability to reduce retrogradation of amylopectin. We have studied its enzymatic properties at pH 5.0. We make two main conclusions: (1) Novamyl^® shows sequence homology to cycloglycosyl transferases (CGTases); like these enzymes, Novamyl^® cleaves cyclodextrins, forms transglycosylation products and is subject to product inhibition by maltose. Novamyl^® has 5 subsites in the active site and is also subject to substrate inhibition. (2) Novamyl^® is clearly different from exoglucanases like β-amylase and glucoamylase. It is able to hydrolyse a pentasaccharide with bulky substituents at both ends (INdp5) and is inhibited by the α-amylase inhibitor Trestatin A. Although Novamyl^® appears unable to hydrolyse α-1,6-linkages, it is able to degrade amylopectin to a greater extent than β-amylase as well as β-limit dextrins. Novamyl^® degrades amylose in such a manner that initially the molecular weight is drastically reduced while β-amylase does not show any detectable effect on the molecular weight of this substrate. Products of the degradation of amylopectin and amylose by Novamyl^® are maltose and oligosaccharides, whereas β-amylase and glucoamylase produce only maltose and glucose, respectively. This was shown in baking experiments as well. The new data presented here clearly show that unlike exoamylases, Novamyl^® does not require a non-reducing end and attacks amylose, Indp5 and cyclodextrins in an endo-like manner. Based on these results Novamyl^® should be reclassified.     

Isolation of obligate anaerobic rumen bacteria capable of degrading the neurotoxin β‐ODAP (β‐N‐oxalyl‐L‐α,β‐diaminopropionic acid) as evaluated by a liquid chromatography/biosensor analysis system

Six pure strains of obligate anaerobes capable of degrading the toxin β‐N‐oxalyl‐L ‐α, β‐diaminopropionic acid (β‐ODAP) contained in grass pea (Lathyrus sativus) have been isolated from cow rumen. The new isolates were identified as Megasphaera elsdenii (five different genotypes) and Clostridium bifermentans using 16S rDNA analysis. The β‐ODAP degrading efficiency of the isolates was evaluated by measuring the amount of β‐ODAP in the growth medium, which contained β‐ODAP as the only carbon source, before and after incubation with the microbes. The method of analysis was liquid chromatography employing bioelectrochemical detection. The biosensor is based on co‐immobilising two enzymes, glutamate oxidase (GlOx) and horseradish peroxidase (HRP), on the end of a spectrographic graphite electrode. β‐ODAP is oxidised by GlOx to form H₂O₂, which in turn is bioelectrocatalytically reduced by HRP through a mediated reaction using a polymeric mediator incorporating Os^{2 + /3+} functionalities rapidly shuttling electrons with the electrode_giving rise to the analytical signal. On the basis of this analysis system, the new isolates are capable of utilising β‐ODAP as sole carbon source to a maximum of 90–95% within 5 days with concomitant increase in cell protein. Copyright © 2005 Society of Chemical Industry     

Comparative study of enzymatic hydrolysis of α/β- and γ-gliadins

Enzymatic hydrolysis of proteins and fractionation of hydrolysates is a route of diversifying their functional properties. Chymotryptic hydrolysis of different sulphur-rich gliadins (α/β- and γ-types), major wheat storage proteins, was studied. The peptides formed in the course of digestion were characterised by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate (SDS-PAGE) and reversed-phase high performance liquid chromatography (RP-HPLC). With reference to previous work, a general scheme of degradation was assessed for γ-gliadins. Limited hydrolysis released two types of polypeptides, comprising respectively the repetitive and the non-repetitive moieties of the protein. In spite of strong sequence homologies between the two groups of sulphur-rich gliadins, it was not possible to prepare similar peptide fractions from α/β-gliadins. They were more resistant to hydrolysis and the region where the two domains merge appeared inaccessible to chymotrypsin. Restricted accessibility of cleavage sites was attributed to the less expanded conformation of α/β-type than γ-type gliadins. A first step of scaling-up was performed. This offers opportunities to prepare functional peptides from wheat storage proteins.     

β-GLUCAN AND β-GLUCANASE IN BREWING

The development of endo β-glucanase during the micro-malting of barley, with and without the addition of gibberellic acid, was compared. Results indicated that the stimulative effect of gibberellic acid on the enzyme system was of only marginal practical importance. From the assessment of the enzyme activity in a number of commercial malts it would appear that the germination time used for some malts is too short to take full advantage of the critical phase of very rapid enzyme development. The viscosity-reducing power of β-glucanase was demonstrated in miniature brewery mashing experiments and details of the progressive degradation of β-glucan by the enzyme system were analysed by gel filtration methods. The β-glucan content of a number of varieties of barley was established.     

Synthesis of β‐Galactooligosaccharides from Lactose Using Microbial β‐Galactosidases

Abstract: Galactooligosaccharides (GOSs) are nondigestible oligosaccharides and are comprised of 2 to 20 molecules of galactose and 1 molecule of glucose. They are recognized as important prebiotics for their stimulation of the proliferation of intestinal lactic acid bacteria and bifidobacteria. Therefore, they beneficially affect the host by selectively stimulating the growth and/or activity of a limited number of gastrointestinal microbes (probiotics) that confer health benefits. Prebiotics and probiotics have only recently been recognized as contributors to human health. A GOS can be produced by a series of enzymatic reactions catalyzed by β‐galactosidase, where the glycosyl group of one or more D‐galactosyl units is transferred onto the D‐galactose moiety of lactose, in a process known as transgalactosylation. Microbes can be used as a source for the β‐galactosidase enzyme or as agents to produce GOS molecules. Commercial β‐galactosidase enzymes also do have a great potential for their use in GOS synthesis. These transgalactosyl reactions, which could find useful application in the dairy as well as the larger food industry, have not been fully exploited. A better understanding of the enzyme reaction as well as improved analytical techniques for GOS measurements are important in achieving this worthwhile objective.     

α 1,4 — α 1,6 Glucopolysaccharides Contained in Developing Maize Kernels

Corn grains with different filling times were fractionated in order to obtain their α 1,4 — α 1,6 glucopolysaccharide composition and structure. Sugary, waxy, flint and several hybrids were the varieties analyzed. There is an increase in the total content of polysaccharides, as the grain is completed. But, no differences in composition and structure were detected between the first sample analyzed (15th day after pollination) and the last one, which corresponds to the complete grain filling period. The methodology developed by us, allowed us to observe that the introduction of su character on the hybrids (F x S and S x F) has influence on the final polysaccharide structure. Also, in the F x W hybridα 1,4 — α 1,6 glucans, their structural characteristics were different from those of F or W pure.     

Investigation of selected toxicological parameters of γ-pentachlorocyclohexene (γ-PCCH)

The toxic effects of the γ-hexachlorocyclohexane metabolite γ-pentachlorocyclohexene (γ-PCCH) were studied by acute and subacute (6 weeks) experiments. The investigations included cerebral convulsibility with chemoshock (Tetrazolium), reactivity with hot plate method, the learning ability with learning tests, and peripheral nervous activity (EMG). Nociceptive reaction time was not influenced, the learning process (6 weeks) was inhibited by γ-PCCH. The conduction velocity of the peripheral nerve was decreased. At the end of the 6th week liver enlargement was found.     

DETERMINATION OF α- AND β-AMYLASE IN MALT

The Analysis Committee of the European Brewery Convention carried out a collaborative trial on malts using the specific analysis methods for α- and β-amylase activities based on dyed substrates supplied by MegaZyme (Aust.) Pty. Ltd. The repeatability and reproducibility values for the methods were judged to be unsatisfactory and consequently the methods were not recommended for Analytica-EBC.     

Purification and physicochemical characterization of ovine β‐lactoglobulin and α‐lactalbumin

Ovine whey proteins were fractionated and studied by using different analytical techniques. Anion‐exchange chromatography and reversed‐phase high‐performance liquid chromatography (HPLC) showed the presence of two fractions of β‐lactoglobulin but only one of α‐lactalbumin. Gel permeation and sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis allowed the calculation of the apparent molecular mass of each component, while HPLC coupled to electrospray ionisation‐mass spectrometry (ESI‐MS) technique, giving the exact molecular masses, demonstrated the presence of two variants A and B of ovine β‐lactoglobulin. Amino acid compositions of the two variants of β‐lactoglobulin differed only in their His and Tyr contents. Circular dichroism spectroscopy profiles showed pH conformation changes of each component. The thermograms of the different whey protein components showed a higher heat resistance of β‐lactoglobulin A compared to β‐lactoglobulin B at pH 2, and indicated high instability of ovine α‐lactalbumin at this pH.     

β-GLUCAN AND β-GLUCAN SOLUBILASE IN MALTING AND MASHING

During malting the water-insoluble β-glucan of barley is diminished whilst water-soluble gum is little decreased. The amount of β-glucan surviving into malt depends on variety but barleys rich in glucan give malts with high β-glucan levels. The β-glucan content of barley depends on variety and growth site. β-Glucan solubilase survives mashing and catalyses the release of hemicellulose into solution. There is no correlation between the β-glucan content of malt and the amount released into wort. However, barley adjuncts containing high levels of β-glucan give worts rich in β-glucan. β-Glucan dissolution in mashing is dependent on time, temperature, grist particle size and liquor: grist ratio. Use of adjuncts derived from barley contribute relatively more β-glucan in wort, coinciding with reduced rates of wort separation, but these can be increased by using a β-glucanase produced by growing the fungus Trichoderma viride on spent grains.     

α-D-Polyglucane-Iodine Complexes

The temperature-dependent dyeability of oligoglucanes and polyglucanes with I₂/KI solution is based on the interaction between iodine and the C-atoms of glucosidic bonds (Vetter and Thorn [1, 2]). Only when the helix is compressed, the expansion of the lumen that is necessary for the attachment of iodine bands is achieved as suggested by Freudenberg et al. in 1939 [3]. The development of colour starts with the maltododecaosis (just over two spirals) within a temperature range of 20 to 25°C adsorbing one pentaiodide and bringing about a pale shade of pink. The rich blue tone is achieved with pure amyloses forming a maximum of about 610 to 620 nm. Unramified polysaccharides which had been synthesised by our team from 30 glucoside residues, still showed a purple colour with a maximum of 510 nm at temperatures from 20 to 25°C. Defined polysaccharides from 40 residues are bluish-purple (just 7 spirals of a tension-free helix) with a maximum absorption of about 550 nm. According to our knowledge, the rich blue tone requires at least 50 glucoside residues linked in an unramified chain [1, 2]. Amylopectin from potatoes has a maximum of 575 to 580 nm which shifts to 555 to 560 nm after partial break-down into β-dextrin by β-amylase (EC 3.2.1.2) and does a long-wave re-shift to 570 nm after splitting off of the side-chain stumps in the basic structure by pullulanase (EC 3.2.1.41). In the case of maize, the maximum is 535 nm for amylopectin, 520 nm for β-dextrin, and 555 nm for the basic structure. This means that in amylopectin from potatoes the helical sections available for the adsorption of iodine bands are longer than those in amylopectin from maize [4]. In glycogen iodine accumulates in a diffuse manner without forming any long bands. For this reason, an absorption shoulder of merely 400 to 500 nm is found by photometry [4]. A treatment with β-amylase makes the situation even worse; only when the side-chain stumps are separated by pullulanase does the formation of iodine bands in the basic structure of the glycogen improve to a maximum of 500 nm. This maximum corresponds to unramified sections of less than 30 glucoside residues [4, 2, 6].     

Cystathionine γ-lyase of Saccharomyces cerevisiae: Structural gene and cystathionine γ-synthase activity

Purification of Saccharomyces cerevisiae cystathionine γ-lyase (γ-CTLase) was hampered by the presence of a protein migrating very close to it in various types of column chromatography. The enzyme and the contaminant were nevertheless separated by polyacrylamide gel electrophoresis. N-terminal amino acid sequence analysis indicated that they are coded for by CYS3(CYI1) and MET17(MET25), respectively, leading to the conclusion that CYS3 is the structural gene for γ-CTLase and that the contaminant is O-acetylserine/O-acetylhomoserine sulfhydrylase (OAS/OAH SHLase). Based on these findings, we purified γ-CTLase by the following strategy: (1) extraction of OAS/OAH SHLase from a CYS3-disrupted strain; (2) preparation of antiserum against it; (3) identification of a strain devoid of the OAS/OAH SHLase protein using this antiserum; and (4) extraction of γ-CTLase from this strain. Purified γ-CTLase had cystathionine γ-synthase (γ-CTSase) activity if O-succinylhomoserine, but not O-acetylhomoserine, was used as substrate. From this notion we discuss the evolutional relationship between S. cerevisiae γ-CTLase and Escherichia coli γ-CTSase.