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.     

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.     

Effect of γ-irradiation on soya bean proteins

The properties of soya bean proteins following γ-irradiation (5–20 kGy) were investigated in connection with protein denaturation. Irradiation doses above 10 kGy caused a decrease in 7S and 11S components and an increase in 2S and 15S components (P < 0.05). However, subunit patterns determined by electrophoresis were not changed appreciably over the entire irradiation dose range. In the differential scanning calorimetry thermogram, the denaturation temperatures of 11S and 7S components were not affected by γ-irradiation, while increased irradiation dose caused a decrease in the enthalpy values of 11S and 7S components due to protein denaturation. Changes in the circular dichroism spectra and tryptophan fluorescence intensity (P < 0.05) were observed only at 20 kGy.     

Effect of γ-irradiation on enzymatic digestion of oil palm empty fruit bunch

Effect of γ-irradiation and combination treatment with alkali on oil palm empty fruit bunch (EFB) was studied to enhance the efficiency of enzymatic digestibility. γ-Irradiation enhanced the enzymatic digestibility, and the amounts of released galacturonic acid and neutral sugars from EFB by Driselase were increased with an increase in irradiation dose. The main components of neutral sugars released from EFB were glucose and xylose. EFB treated with alkali and 50 kGy irradiation released 1200 μg g^?1 of xylose that was twice amount of glucose showing that the combination treatment was effective especially to increase the digestibility of xylan.     

Vergleich der Verteilung und Toxizität von α-, β- und γ-Hexachlorcyclohexan (HCH) nach 30tägiger Verabreichung an Ratten

Administration of 1000 ppm α-HCH, 600 ppm β-HCH and β 125 ppm γ-HCH to rats caused a growth retardation. α-HCH and β-HCH led to livermass enlargement. β-HCH caused a decrease of absolute brainmass. In case of α-HCH and γ-HCH fat tissue, kidneys and adrenals had a high burden in residues. Fat tissue and adrenals had high residues of β-HCH. The level of HCH-residues during the test period was γ ≦ β < α. The results are discussed taking into account the contradictory knowledge from literature.     

Chlorinated residues in the adipose tissue of pigs treated with γ-hexachlorocyclohexane

Experiments were conducted to study the metabolic fate of γ-HCH used in pig husbandry, and to see if a relationship existed between its use and the occurrence of 1,4-dichlorobenzene and other chlorobenzenes in pork meat. Both oral administration and spray application of γ-HCH led to the rapid accumulation of γ-HCH and several metabolites in the adipose tissue. The following compounds were identified by gas chromatography and mass spectrometry: monochlorobenzene, 1,2-, 1,3- and 1,4-dichlorobenzene, 1,2,4-trichlorobenzene, 1,2,4,5-, 1,2,3,5- and 1,2,3,4-tetrachlorobenzene, pentachlorobenzene, γ-3,4,5,6-tetrachlorocyclohexene, γ-2,3,4,5,6-penta-chlorocyclohexene and α-hexachlorocyclohexane. Concentrations of residual γ-HCH and its metabolites were related to the amount of γ-HCH applied initially to the pigs. γ-HCH residues and the metabolites were monitored by biopsy and were found to be eliminated rapidly from the adipose tissue, such that pigs sprayed once with a dose 16 times greater than the recommended rate had a residual γHCH concentration, 30 days after treatment, no greater than that in untreated controls. The origin of 1,4-dichlorobenzene and other chlorobenzene compounds is discussed in relation to the low levels of these compounds often found in flavour extracts of pork meat.     

Investigation of selected toxicological parameters of gamma-pentachlorocyclohexene (gamma-PCCH).

The toxic effects of the gamma-hexachlorocyclohexane metabolite gamma-pentachlorocyclohexene (gamma-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 gamma-PCCH. The conduction velocity of the peripheral nerve was decreased. At the end of the 6th week liver enlargement was found.     

Influence of packaging atmosphere on the formation of cholesterol oxides in γ-irradiated egg powder

In a previous work the effect of ionising radiation on formation of cholesterol oxidation products (COPs) under aerobic conditions was studied. In the present work the influence of aerobic and anoxic conditions have been comparatively investigated to study the chemical changes of cholesterol in γ-irradiated egg powder. The irradiation treatment was carried out with powdered egg packed under air and also under vacuum in polyethylene (PE) bags and in laminated, oxygen impermeable three-layer (polyester-aluminium-polyethylene) foil to dosage levels 2 and 4 kGy at room temperature. The cholesterol oxidation is demonstrated by thin-layer chromatograms. In the egg powder wrapped in PE bags the predominant cholesterol derivatives?7-hydroxycholesterol isomers (α and β)—accumulated in significant amounts (increasing with dose) while vacuum-packaging in laminated foil considerably suppressed the quantity of these products and prevented the formation of cholesterol 5α, 6α-epoxide as well as 7-ketocholesterol. Little or no change was observed in non-irradiated (control) vacuum-packed egg powder stored at approximately 22°C for up to 5 months. Peroxide values showed changes parallel to the formation of COPs.     

Comparing effects of α‐ vs. β‐chitosan coating and emulsion coatings on egg quality during room temperature storage

Effects of α‐ and β‐chitosan (CH), soybean oil (SO) and their emulsions (CH:SO = 2:3) as coating materials on selected internal quality and sensory properties of eggs were evaluated during 5 weeks storage at 25 °C. After 3 weeks of storage, α‐ and β‐CH‐coated eggs changed to B grade, while SO‐ and emulsion‐coated eggs preserved grade A quality. Weight loss of eggs coated with SO and CH:SO emulsions was <2.0% vs. 5.3–5.8% for noncoated and CH‐coated eggs after 5 weeks of storage. β‐CH (0.9%) maintained lower weight loss of eggs than α‐CH (1.2%) only at 1‐week storage. Albumen pH of eggs coated with SO and CH:SO emulsions decreased progressively throughout storage. Eggs coated with β‐CH:SO emulsion and SO were significantly glossier than noncoated eggs. Consumers indicated positive purchase intent (69.17–76.67%) for all coated eggs. Overall, α‐CH:SO and β‐CH:SO emulsions extended egg shelf life by at least 3 weeks during room temperature storage.     

Preparation of Recombined Milk Using Modified Butterfats Containing α‐Linolenic Acid

Abstract: Modified butterfats (MBFs) were produced by lipase‐catalyzed interesterification with 2 substrate blends (6:6:8 and 4:6:10, by weight) of anhydrous butterfat (ABF), palm stearin, and flaxseed oil in a stirred‐batch type reactor after short path distillation. The 6:6:8 and 4:6:10 MBF contained 21.7% and 26.5%α‐linolenic acid, respectively. Total saturated fatty acids of the MBFs ranged from 41.4% to 47.4%. The cholesterol contents of the 6:6:8 and 4:6:10 MBFs were 21.0 and 12.1 mg/100 g, respectively. In addition, the melting points of the 6:6:8 and 4:6:10 MBFs were 32 °C and 31 °C, respectively. After preparation of recombined milks (oil‐in‐water emulsions) with MBFs, the stability of emulsions prepared with the MBFs (6:6:8 and 4:6:10) was compared to those with ABF during 10‐d storage at 30 °C. Skim milk powder (containing 1% protein) was added to prepare emulsions as an emulsifier. Microstructures of emulsions freshly prepared with the ABF and the MBFs consisted of uniform fat globules with no flocculation during 10‐d storage. With respect to fat globule size distribution, the volume‐surface mean droplet diameter (d₃₂) of the 6:6:8 and 4:6:10 MBF emulsions ranged between 0.33 and 0.34 μm, which was similar to the distribution in ABF emulsion. Practical Application: Milk, an expensive dairy food, has been widely used in various milk‐derived food products. Modified butterfats (MBFs) contain α‐linolenic acid as an essential fatty acid. Emulsion stability of recombined milks (oil‐in‐water emulsions) with MBFs was similar to that in anhydrous butterfat emulsion during 10‐d storage. They may be a promising alternative for reconstituted milks to use in processed milk‐based products.     

Characterization of the Supermolecular Structure of Polydatin/6‐O‐α‐Maltosyl‐β‐cyclodextrin Inclusion Complex

Polydatin is the main bioactive ingredient in many medicinal plants, such as Hu‐zhang (Polygonum cuspidatum), with many bioactivities. However, its poor aqueous solubility restricts its application in functional food. In this work, 6‐O‐α‐Maltosyl‐β‐cyclodextrin (Malt‐β‐CD), a new kind of β‐CD derivative was used to enhance the aqueous solubility and stability of polydatin by forming the inclusion complex. The phase solubility study showed that polydatin and Malt‐β‐CD could form the complex with the stoichiometric ratio of 1:1. The supermolecular structure of the polydatin/Malt‐β‐CD complex was characterized by ultraviolet–visible spectroscopy (UV), Fourier transform infrared spectroscopy (FT‐IR), X‐ray diffractometry (XRD), thermogravimetric/differential scanning calorimetry (TG/DSC), and proton nuclear magnetic resonance (¹H‐NMR) spectroscopy. The changes of the characteristic spectral and thermal properties of polydatin suggested that polydatin could entrap inside the cavity of Malt‐β‐CD. Furthermore, to reasonably understand the complexation mode, the supermolecular structure of polydatin/Malt‐β‐CD inclusion complex was postulated by a molecular docking method based on Autodock 4.2.3. It was clearly observed that the ring B of polydatin oriented toward the narrow rim of Malt‐β‐CD with ring A and glucosyl group practically exposed to the wide rim by hydrogen bonding, which was in a good agreement with the spectral data.     

PRODUCTION OF BEER USING IMMOBILIZED YEAST ENCODING α-ACETOLACTATE DECARBOXYLASE

Genetically modified brewer's yeast encoding α-acetolactate decarboxylase (α-ALDC) was tested in immobilized yeast bioreactors for main fermentation of beer. The α-ALDC enzyme produced by the transformant catalyzes the direct conversion of α-acetolactate to acetoin without formation of diacetyl. The long lagering period required for beer maturation in conventional brewing can thus be shortened or even omitted. Three different packed bed bioreactors were employed, with volumes of 1.6 dm³, 5 dm³ and 25 dm³. The 5 dm³ column had a slightly conical geometry in contrast to the others which had cylindrical shapes. Sintered glass beads were chosen as the carrier material on the basis of experiments with the parent strain. The brewing performance of the transformant compared well with that of the parent strain in the immobilized system. Fermentation, utilization of amino acids (including isoleucine, valine and leucine) and flavour formation were practically identical with both strains, the only difference being a marked decrease in the formation of diacetyl by the transformant. Small differences were, however, observed in the long-term biochemical stability. By using yeast encoding α-ALDC in the immobilized yeast system the total (primary and secondary) fermentation time could be reduced to approximately 2–6 days, compared with 3–6 weeks in a conventional batch process.     

Whitening effect of α‐bisabolol in Asian women subjects

Although skin pigmentation, which results from the production and distribution of melanin in the epidermis, is the major physiological defence against solar irradiation, hyperpigmentation is a common and distressing problem caused by various inflammatory skin disorders, such as eczema, allergic contact dermatitis and irritant contact dermatitis. We evaluated the effects of a preparation containing α‐bisabolol on pigmented skin of a group of subjects. The effectiveness of the active compound, α‐bisabolol, in a base‐cream preparation for the treatment of pigmented skin was tested on 28 female subjects as follows: the cream was applied once a day to the back for 8 weeks. These same women also applied a vehicle control cream to the pigmented skin. The results were evaluated by clinical and biophysical test methods. After 8 weeks of treatment of the α‐bisabolol‐containing cream, there was significant lightening effect in the pigmented skin for the majority of the subjects who tested the cream.     

Properties of soya milk and tofu prepared with γ-irradiated soya beans

Soya beans (Glycine max) ‘hwang keum’ were γ-irradiated at dose levels of 0, 2.5, 5, 10 and 20 kGy and the effects of the irradiated soya beans on soya milk and tofu properties were studied. An irradiation dose of 5 kGy caused an increase in yield of soya milk and tofu while having very little effect on their quality. The properties of tofu prepared with the soya beans irradiated at 2.5–5 kGy showed no significant difference from the non-irradiated control. However, at higher doses (10–20 kGy), decreases in yield, water holding capacity and sag value of tofu were observed. Compared with the non-irradiated control, hardness and fracturability in the texture of tofu were both significantly increased when the soya bean had been irradiated at 10–20 kGy, while cohesiveness and adhesiveness decreased. The changes in color values of soya milk and tofu were pronounced at 20 kGy.     

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.     

Adsorption of Shiga Toxin to Poly‐γ‐Glutamate Precipitated

We screened foods containing indigestible ingredients in the ability to adsorb Shiga toxin (Stx). When 5 mg of foods and dietary fibers such as dry vegetables and inulin were mixed and incubated with 0.5 mL of Stx solution (100 ng/mL) containing 0.5% bovine serum albumin, both Stx1 and Stx2 seemed to be adsorbed by only a fermented food, natto (a traditional Japanese food prepared from steamed soybeans by the biological action of Bacillus subtilis). We purified the Stx‐adsorbing substance from natto by extraction with H₂O, acid treatment, Proteinase K treatment, and an ion exchange chromatography. The purified substance showed an average molecular mass of about 600 kDa. We identified it as poly‐γ‐glutamate (PGA) by amino acid analysis of its hydrolysate and peptide analysis after its treatment with Proteinase K. Purified PGA (MW: molecular weight = about 600 kDa) was considered to adsorb both Stx1 and Stx2 when we separated adsorbed and unadsorbed Stxs (MW = about 72 kDa) by an ultrafiltration method with a centrifugal filter unit (MWCO: molecular weight cut‐off = 100 K). However, PGA with the ability to adsorb Stx was an insoluble form precipitated in the filter unit during centrifugation. PGA precipitated beyond the saturated density was also confirmed to well adsorb both Stx1 and Stx2 by an equilibrated dialysis method. To the best of our knowledge, this is the 1st report on food‐adsorbing Stx.     

Bioequivalence of β‐carotene and retinol

For many years it was accepted that 6 mg of β‐carotene were required to produce 1 mg of vitamin A in the form of retinol. The equivalence was based on the assumptions that two‐thirds of dietary β‐carotene are not absorbed, while in the metabolism of the remaining third 1 mol of β‐carotene is converted to 1 mol retinol. Recently, the bioequivalence was raised to 12 mg β‐carotene and 1 mg retinol. The objective of this review was to re‐examine the data that were used to support the new equivalence ratio, especially since some of these data were obtained in developing countries where infestation with gut parasites and exposure to other infections is common, yet the influence of inflammation on plasma carotenoid and retinol concentrations is frequently ignored. Bioequivalence studies examined in this review include those done in developing and developed countries, depletion and repletion studies, feeding with vegetable sources of β‐carotene or pure supplements, influence of helminths, carotenoid interactions and matrix effects and studies using stable isotopes (SI). SI studies show the bioefficacy of β‐carotene conversion to retinol is generally poor even for pure β‐carotene unless the dose is small and fed regularly until equilibration is reached. Retinol formation appears to be inversely influenced by previous vitamin A intake, the amount of material given and current vitamin A status. In spite of technical complexities, more SI studies where liver reserves of vitamin A are determined pre and post intervention are needed to evaluate β‐carotene bioefficacy of different vegetable sources. Copyright © 2006 Society of Chemical Industry     

Effects of poly‐γ‐glutamic acid on the physicochemical characteristics of skim milk yoghurt

The physicochemical and sensory properties of skim milk yoghurts containing poly‐γ‐glutamic acid (PGA) at different levels (0.0025, 0.005 and 0.01%) were evaluated. Addition of PGA up to 0.01% to reconstituted skim milk (11%, w/v) did not affect the growth of lactic acid bacteria or the development of titratable acidity in yoghurt, whereas full‐fat control yoghurt had reduced acid production. No changes were found in viable cell counts of PGA yoghurts during storage (4 weeks at 4 °C). The addition of PGA (0.005%) significantly decreased syneresis in skim milk yoghurt and did not cause any undesirable effects in sensory acceptability.