Hydrolysate Research Articles

Management of gastrointestinal food allergy in childhood.

Gastrointestinal food allergy in childhood is characterised by onset of gastrointestinal symptoms following food ingestion where the underlying mechanism is an immunologically mediated reaction within gastrointestinal tract. Presentation may be quick, slow or quick and slow after food ingestion. Diagnosis depends on response to food elimination, response to food challenge and analysis of response to food elimination. Management centres upon an elimination diet, the need for this is temporary. Cow's milk protein hydrolysates or amino acid formulae are preferred to soy formulae.

Conversion of wax apple and taro stalk wastes to ethanol by genetically engineered escherichia coli strains

Recombinant strains of Escherichia coli ST09, ST32, and ST44 containing saccharification and fermentation genes were constructed. Pyruvate decarboxylase and alcohol dehydrogenase genes from Zymomonas mobilis and amylase gene from Bacillus licheniformis were integrated into the chromosome of E. coli. Endoglucanase gene from Clostridium cellulovorans was maintained on a plasmid pEB4 derived from pET8c. These genetically recombinant novel strains could convert 10% glucose to 4.9–6.1% (v/v) ethanol, which was equivalent to 77–96% of the theoretical yield in 72 h. Fermentation of blended wax apple and taro hydro lysate mix (10% reducing sugar equivalent) by ST44 was essentially complete after 72h with final ethanol concentrations of 4.2 and 3.8%(v/v), respectively.

The use of radiolabelling techniques to measure substantivity to, and penetration into, hair of protein hydrolysates

The use of 14C-labelled amino acids enables the measurement of both the total substantivity to hair and the degree of penetration into the hair shaft of amino acid mixtures derived from complete hydrolysis of proteins. The technique utilizes the fact that direct measurement of 14C radioactivity of the treated hair detects only the surface substantivity. Total substantivity can be determined following solubilization of the hair. Data obtained for wheat amino acids show significant penetration when used to treat hair from a shampoo or conditioner formulation. A similar technique has been investigated for a wheat protein partial hydrolysate using 14CNO for radiolabelling purposes and shows that significant penetration into hair can occur. L'utilisation d'amino-acides marques au 14C permet la mesure a la fois de l'absorption totale par les cheveux et du degre de penetration dans la fibre du cheveu de melanges d'amino-acides obtenus a partir d'une hydrolyse totale de proteines. La technique utilise le fait que la mesure directe de la radioactivite du 14C des cheveux traites ne detecte que l'absorption en surface. L'absorption totale peut etre determinee apres solubilisation des cheveux. Les donnees obtenues a partir d'amino-acides de ble montrent une penetration significative lors d'une utilisation pour traiter des cheveux a partir d'une formulation de shampoing ou d'apres-shampoing. Une technique similaire a ete exploree vis-a-vis d'un hydrolysat partiel d'une proteine de ble utilisant 14CNO a des fins de marquage, et montre qu'une penetration significative dans les cheveux peut avoir lieu.

Isolation from bovine haemoglobin of a peptide that might be used as a potential hydrophobic photosensitizer carrier.

The isolation of a peptic hydrolysate and of a pure peptide from bovine haemoglobin is described. These peptides were used to solubilize an insoluble photosensitizer in order to study their utilization as a carrier for photochemotherapy. Protoporphyrin IX was used as a model of an insoluble photosensitizer. Solubilization of protoporphyrin IX was first performed with the total haemoglobin hydrolysate, then with peptide fractions, and finally with a pure peptide isolated from this fraction by reversed-phase HPLC. The molecular mass and the primary structure of the pure peptide were determined by fast-atom-bombardment and tandem MS (molecular mass 1648 Da). The singlet oxygen quantum yield of the protoporphyrin IX-peptide complex was determined.

Production of xylitol from raw wood hydrolysates by Debaryomyces hansenii NRRL Y-7426

Production of xylitol from raw wood hydrolysates by Debaryomyces hansenii NRRL Y-7426

Effect of the aminopeptidase from Pseudomonas fluorescens ATCC 948 on synthetic bitter peptides, bitter hydrolysate of UHT milk proteins and on the ripening of Italian Caciotta type cheese

Effet de l'aminopeptidase de Pseudomonas fluorescens ATCC 948 sur les peptides amers synthetiques, les hydrolysats de proteines du lait UHT et sur l'affinage du fromage italien de type caciotta. On a etudie l'activite de l'aminopeptidase purifiee de Pseudomonas fluorescens ATCC 948 sur les peptides amers synthetiques, les hydrolysats amers des proteines du lait UHT et sur l'affinage du fromage italien de type caciotta. L'aminopeptidase a presque totalement hydrolyse le pentapeptide amer H-Leu-Trp-Met-Arg-Phe-OH et libere Val du tetrapeptide amer H-Val-Pro-Leu-Leu-OH. Aucun clivage de la liaison Pro n'a ete observe. Dans le lait UHT, dans lequel l'hydrolyse des proteines a ete provoquee par addition d'une proteinase, l'aminopeptidase a produit une concentration d'acides amines libres environ 8 fois plus elevee que celles determinees sur le lait UHT non hydrolyse et non amer (211,3 contre 25,3 μg/ml respectivement). Glu, Leu, Met, Trp et Val ont ete les plus abondants. L'enzyme a presente 44% de son activite maximale a la temperature de conservation du lait UHT (20°C). L'aminopeptidase a ete stable pendant l'affinage de la caciotta : aucune activite n'a ete perdue pendant 2 mois. Au terme de 45 jours d'affinage, le fromage contenant l'aminopeptidase presentait un niveau d'acides amines plus eleve (990,2 μg/ml) que le controle non traite (591,1 μg/ml). Le profil des acides amines du fromage traite refletait l'activite de l'aminopeptidase. L'hydrolyse specifique des liaisons peptidiques impliquant des acides amines (Leu, Trp et Val) habituellement identifies comme etant les principaux composants des peptides amers indique probablement une activite d'elimination de l'amertume de cette aminopeptidase.

Evidence for α-1,6 and α-1,4-glucosidic bond cleavage in highly branched glycogen by amylopullulanase from Thermoanaerobacter ethanolicus

Activity of amylopullulanase from Thermoanaerobacter ethanolicus 39E on α-1,6 and α-1,4-glucosidic linkages in highly branched mammalian glycogen was analyzed by paper chromatography and 13C nuclear magnetic resonance (NMR) spectroscopy. Paper chromatography analysis showed that the glycogen hydrolysate consisted of glucose, maltose, maltotriose and maltotetraose. NMR spectroscopy confirmed that no hydrolysate products of α-1,6 linkage were present resulting from treatment with the amylopullulanase. Therefore, the amylopullulanase efficiently hydrolyzed glycogen both at α-1,6- and at α-1,4-glucosidic linkages into oligosaccharides.

Modelling of Continuous Corn Starch Hydrolysate Fermentations in Immobilised Cell Reactors

AbstractCorn starch hydrolysate has been fermented by two different strains of Saccharomyces cerevisiae and Saccharomyces oviformis, either in batch or in continuous modes, using both suspended‐cell and en‐trapped‐cell systems, provided with different porous supports, namely a commercial synthetic sponge, crushed almond skins and alginate beads. The present results demonstrate that starch hydrolysate is an excellent substrate for alcohol fermentation, whether out of the presence of growth factors for the yeast or in absence of inhibiting substances.A semi‐empirical kinetic model, previously proposed for comparing the results of continuous fermentations of various substrates in different reactors, is now successfully extended to the study of different porous supports for cell entrapment in a fixed‐bed column. The best results in terms of ethanol productivity (7.8 gp/1h, referred to the whole reactor volume) are obtained entrapping the yeast cells within the spongy matrix.

Purification and properties of acetylcholinesterase from human brain.

Acetylcholinesterase from human caudate nucleus and partial thalamus was purified by using Con A-Sepharose, short-arm and long-arm ligand Sepharose affinity chromatographies. SDS-PAGE of the purified AChE under the reduced condition showed one main band, corresponding to a molecular weight of 66 kD. The purified AChE with a specific activity of 3384 U/mg protein represented 20% activity of the homogenate supernatant. Analysis of purified AChE by gradient slab PAGE and DISC-PAGE with activity staining revealed the existence of monomer, dimer, tetramer, hexamer and octomer of the enzyme. The isoelectric point of AChE ranged between pH 5.6 and 6.0. Con A-Sepharose affinity chromatography retained most of the applied AChE activity implying that the enzyme is a kind of glycoprotein. The isolated human brain AChE had no cross-immunoreactivity with 3F3 and weak cross-immunoreactivity with 2G8 and 1H11 anti-Torpedo AChE antibodies. Balb/c mice were immunized with human cerebellum AChE purified with Con A and short-arm ligand affinity chromatographies. The antiserum produced showed strong cross-immunoreactivity with Torpedo AChE but weak cross-immunoreactivity with human RBC membrane AChE. The purified human brain striatum AChE was reduced and alkylated, and then hydrolyzed by immobilized TPCK-treated trypsin. Trypsin peptides in the hydrolysate was separated by RP-HPLC. Several large peptide peaks and numbers of small peaks were observed. The large peaks showed obvious immunoreactivity with the mouse anti human cerebellum AChE antiserum.

Effect of fucoidin on human sperm-zona pellucida interactions.

The authors recently reported that fucoidin (a polymer of predominantly L-fucose sulfate) produced a strong, significant, and dose-dependent inhibition of sperm-zona binding under hemizona assay conditions. The current studies were designed to evaluate the mechanisms underlying this inhibitory activity. Using computerized semen analysis, the monoclonal anti-sperm antibody T-6 and indirect immunofluorescence technique, and the fura-2 indicator, no significant impact of fucoidin on sperm motion parameters, on the spontaneous acrosome reaction, or its prerequisite, the increase in calcium influx, were observed. Subsequently, a mild acid hydrolysis of the fucoidin molecule was performed, followed by sizing hydrolysates in a Biogel P2 column and separating fractions (n = 6). All fucoidin fragments significantly inhibited tight binding of human sperm to human zona pellucida under hemizona assay conditions (range, 56%-94%) when tested individually. These results provide further evidence that the effect of fucoidin is produced by a receptor-ligand association.

Alpha-Chymotrypsin in plastein synthesis: influence of substrate concentration on enzyme activity.

The role of alpha-chymotrypsin in the plastein reaction was studied using a peptic hydrolysate of albumin as substrate. Study of this reaction simultaneously by different methods showed that the plastein reaction is enzyme catalyzed and is highly dependent on environmental conditions. A gel permeation chromatography study of the plastein reaction showed simultaneous increases in the high- and low-molecular-weight oligopeptide fractions; a transpeptidation mechanism may be involved in the reaction. A study of the effect of substrate concentration on the plastein reaction catalyzed by alpha-chymotrypsin showed a profile with both hydrolytic and synthetic activities. This effect was also observed when the reaction course was followed by quantification of the free amino groups at different substrate concentrations, showing that a condensation mechanism is responsible for the synthetic activity when the substrate concentration is very high. These results have led us to conclude that the plastein reaction involves a transpeptidation and/or condensation mechanism, which is a function of the substrate concentration.

DIRECT SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM PROTOPLAST-DERIVED CELLS OF WHEAT ( Triticum aestivum L.)

Protoplasts were isolated from embryogenic calli derived from immature embryos of wheat (Triticum aestivum L. cv. Jinan 177). The protoplasts were cultured in NMB medium supplemented with 1mg/L 2,4- D and 500mg/l casein hydrolysate (CH). The regenerated cells from protoplasts divided to form somatic embryos directly. The somatic embryos grown to 1.5- 2 mm in size directly developed into complete plants on solid MB medium without hor- mones.

Analysis of bacterial exopolysaccharides.

Extracellular polysaccharides have been isolated from cultures of freshwater and marine bacteria originally isolated from material adhering to surfaces and underivatized hydrolysates have been analyzed by high-performance liquid chromatography methods. A scheme has been developed whereby the uronic acids can be identified on strong anion-exchange columns, while neutral monosaccharides can be separated and identified using aminobonded columns or cation-exchange adsorbent loaded with a heavy metal ion. The methods permit rapid and accurate comparison of polysaccharides with differing chemotype. The strains studied show a range of different chemotypes, all containing a uronic acid and several neutral monosaccharides. Some of the polysaccharides isolated from marine bacteria possessed a very high acetyl content.

Capillary gas chromatography of neutral sugars as their aldononitrile acetates from the hydrolyzate of corn bran residues

Presentation d'une methode d'analyse de la fraction glucidique de ce sous-produit d'amidonnerie de mais

Preparative high-performance liquid chromatography of malto-oligosaccharides

New preparative high-performance liquid chromatographic techniques that allow the practical isolation of milligram to gram quantities of malto-oligosaccharides from starch hydrolysates are described. High-resolution fractionations were achieved on preparative ( − 30 × 2.0 cm) columns packed with either (i) H +-form cation-exchange resin, (ii) Ag +-form cation-exchange resin, (iii) aminopropyl silica gel, or (iv) octadecyl (C 18) silica gel. Details are given for the preparation and packing of the first three phases and for the practical use of all four for rapid oligosaccharide fractionations. Although each column had some unique features, they all operated at surprisingly low flow-rates (2–12 ml/min) and pressures (15–1000 p.s.i.). In addition, most were relatively easy and inexpensive to assemble and operate. The use of these columns on standard analytical-scale chromatographs and also on automated preparative high-performance liquid chromatography systems, for the isolation of the title compounds, is described.

Urinary excretion of desmosine (elastin cross-links) in subjects with PiZZ alpha-1-antitrypsin deficiency, a phenotype associated with hereditary predisposition to pulmonary emphysema.

To evaluate the concept that lung elastin degradation is accelerated in homozygous alpha-1-antitrypsin (AAT) deficient persons, we prepared acid hydrolysates of urine and used a radioimmunoassay for desmosine to measure urine concentrations of this elastin-specific cross-link in such persons and in control subjects. Excretion of desmosine in 17 homozygous AAT-deficient (PiZZ) patients with emphysema was compared with that in 27 patients with interstitial lung diseases (16 sarcoid, 5 idiopathic pulmonary fibrosis, 6 other interstitial lung diseases) and 26 healthy subjects. Both smokers and nonsmokers were present in all groups. Urinary desmosine concentration (microgram/100 mg creatinine) was 2.35 +/- 0.93 in the PiZZ patients, 2.49 +/- 1.01 in those with interstitial lung disease, and 2.05 +/- 0.54 in the healthy control subjects (p greater than 0.1, all comparisons). Because abnormal pulmonary elastolysis may be largely completed before symptoms of emphysema develop in AAT-deficient persons, we also tested 6 asymptomatic adults with homozygous AAT deficiency (PiZZ) and 5 PiZZ children. Urine desmosine (microgram/100 mg creatinine) was not significantly elevated in either group compared with that in the age-matched control subjects, although children (PiZZ and age-matched controls) showed higher excretions than did adults (6 asymptomatic PiZZ adults, 2.60 +/- 0.91; 5 PiZZ children, 3.27 +/- 0.62; 10 control children, 3.61 +/- 0.62). These data suggest that pathologic lung elastolysis in the PiZZ subject may constitute too small a fraction of total-body elastin turnover to be detected by this method.(ABSTRACT TRUNCATED AT 250 WORDS)

Immobilization of proteins to hydrolyzate lignin by means of formaldehyde

Preparation d'un support peu onereux par utilisation d'un hydrolysat de lignine activee au moyen de formaldehyde en milieu alcalin de preference. Le traitement par le formaldehyde, permet la formation d'un grand nombre de groupes reactifs capables d'immobiliser de grandes quantites de proteines (chymotrypsine, trypsine, ovomucoide)

Secretion of 3,3'-diiodothyronine by the perfused canine thyroid isolated in situ.

Previous studies employing perfused dog thyroid lobes have shown that thyroidal secretion is modulated by intrathyroidal deiodination of T4 to T3 and rT3. To characterize further the intrathyroidal iodothyronine-deiodinating processes 3,3'-diiodothyronine (3,3'-T2) and T4 were measured in hydrolysate and effluent from isolated dog thyroid lobes during single passage perfusion with a synthetic hormone-free-medium. In five experiments the concentrations in effluent from unstimulated thyroid lobes were: 3,3'-T2, 208 +/- 62 pmol/liter; and T4, 12.8 +/- 2.8 nmol/liter (mean +/- SE). During the infusion of 100 microU/ml TSH, there was a parallel increase in the secreton of 3,3'-T2 and T4. After approximately 90 min of TSH infusion, effluent concentrations were: 3,3'-T2, 2490 +/- 420 pmol/liter; and T4, 204 +/- 24 nmol/liter. In a pronase hydrolysate of the thyroid lobes, the amounts were: 3,3'-T2, 2.44 +/- 0.56 pmol/mg; and T4, 1070 +/- 125 pmol/mg. Expressed as a percent of T4, the amount of 3,3'-T2 in thyroid effluent was approximately 10 times higher than that in thyroid hydrolysate. This relative hypersecrection of 3,3'-T2 was abolished by the infusion of 10(-3) or 10(-5) M propylthiouracil, an inhibitor of peripheral and intrathyroidal iodothyronine deiodination. Thus, thyroid secretion contains small amounts of 3,3'-T2, most of which seems to originate from intrathyroidal deiodination of T3 and/or rT3.

An in vitro study on the interaction between dimethylnitrosamine and nucleic acids via a microsomal system.

Radioactive alkylated bases, ribose or phosphate, were never found either in acid and alkaline hydrolysates of polyribonucleotides or in alkaline hydrolysates of DNA after incubation with 14C-dimethylnitrosamine (DMNA) in a microsomal system. Two radioactive compounds, which were co-chromatographed with methylamine and N-methylhydrazine, respectively, on column, paper, and thin-layer, were always detected. They differed from the compound derived from 7-methylguanosine after the alkali-mediated fission of the imidazole ring in its molecule. The in vitro system employed well represents the in vivo situation (7-methylguanine which is liberated from DNA after acid hydrolysis); however, it has given results which do not agree with the generally-accepted mechanism of DMNA alkylation at the N-7 position of guanine.

Antitumor activity of a new antitumor antibiotic, sporamycin.

Sporamycin is an antitumor antibiotic isolated from the culture filtrate of Streptosporangium strain No. PO-357. Hydrolysate of the antibiotic showed at least 12 kinds of amino acid, and the molecular weight was calculated approximately as 8,500 approximately 9,000. Antitumor activities of Sporamycin were examined on murine tumors according to different schedules of treatment. After nine daily doses of the antibiotic had been given to dd mice bearing Ehrlich ascites carcinoma, significant increase of lifespan was observed at doses of 5.0 approximately 1.3 mg/kg. A marked decrease in tumor size of sarcoma-180 transplanted subcutaneously was observed when the mice were treated by daily or single injection of Sporamycin. Against leukemia P-388 and L-1210 inoculated intraperitoneally into CDF1 mice, the antibiotic increased lifespan of leukemic mice. The antibiotic also inhibited significantly the growth of human tumor cells in vitro.