Cytotoxic and allergenic potential of bioactive proteins and peptides.

This review article deals with the assessment of cytotoxic and allergenic potential of bioactive proteins and peptides. It is evident that 'novel' foods or nutraceuticals containing bioactive proteins and peptides must fulfill their proposed "health claim". Furthermore, there is a need to assess their potential to exert adverse effects before they can be made widely available to consumers.

Influence of heat impact in reconstituted skim milk on the properties of yoghurt fermented by ropy or non-ropy starter cultures.

The paper describes studies on the influence of heat impact in reconstituted skim milk on chemical and functional properties of yoghurt products. Reconstituted skim milk was heated for 20 min at 85 degrees C, 90 degrees C, or 95 degrees C. Ropy (producing exopolysaccharides, EPS) or non-ropy strains of S.

Biology of the temperate Streptococcus thermophilus bacteriophage TP-J34 and physical characterization of the phage genome.

The temperate Streptococcus thermophilus bacteriophage TP-J34 was identified in the lysogenic host strain J34. The majority of phage particles produced upon induction was defective and noninfectious, consisting of DNA-filled heads lacking tails. A physical map (45.

Identification and differentiation of species and strains of Arthrobacter and Microbacterium barkeri isolated from smear cheeses with Amplified Ribosmal DNA Restriction Analysis (ARDRA) and pulsed field gel electrophoresis (PFGE).

ARDRA (Amplified Ribosomal-DNA Restriction Analysis) was used to differentiate among species and genera of Arthrobacter and Microbacteria. Species-specific restriction patterns of PCR-products were obtained with NciI for Arthrobacter citreus (DSM 20133T), A. sulfureus (DSM 20167T), A.

Application of the shsp gene, encoding a small heat shock protein, as a food-grade selection marker for lactic acid bacteria.

Plasmid pSt04 of Streptococcus thermophilus contains a gene encoding a protein with homology to small heat shock proteins (A. Geis, H. A.

Sequence analysis and characterization of plasmids from Streptococcus thermophilus.

The nucleotide sequences of eight plasmids isolated from seven Streptococcus thermophilus strains have been determined. Plasmids pSt04, pER1-1, and pJ34 are related and replicate via a rolling circle mechanism. Plasmid pJ34 encodes for a replication initiation protein (RepA) and a small polypeptide with unknown function.

Biofunctional peptides from milk proteins: mineral binding and cytomodulatory effects.

The protein fraction of milk contains many valuable components and biologically active substances. Moreover, milk proteins are precursors of many different biologically active peptides which are inactive within the sequence of the precursor protein but can be released by enzymatic proteolysis. Many milk protein-derived peptides, such as caseinophosphopeptides, reveal multi-functional bioactivities.

Detection of caseinophosphopeptides in the distal ileostomy fluid of human subjects.

Caseinophosphopeptides (CPP) were detected for the first time in ileostomy fluid, collected at 2 h intervals for 10 h post milk and CPP ingestion, from human volunteers with an ileostomy. The level of CPP present in ileostomy fluid obtained from milk-fed volunteers was markedly higher than that from volunteers fed with selected CPP preparations. The findings are based on HPLC analysis in combination with peptide-bound P determination, thin-layer electrophoresis and amino acid analysis, together with ELISA studies using polyclonal antibodies raised against a set of CPP to detect immunoreactive CPP in ileostomy fluid.

Effect of oligofructose or dietary calcium on repeated calcium and phosphorus balances, bone mineralization and trabecular structure in ovariectomized rats*.

We investigated the effects of dietary oligofructose and Ca on bone structure in ovariectomized rats, using microradiography and histomorphometry. Ninety-six animals were allocated to seven experimental groups: G1, sham-operated; G2-G7, ovariectomized. Semi-purified diets containing 5 g Ca/kg (recommended content) without oligofructose (G1, G2) or with 25, 50 or 100 g oligofructose/kg (G3, G4, G5) or 10 g Ca/kg (high content) without oligofructose (G6) or with 50 g oligofructose/kg (G7) were fed for 16 weeks.

Probiotics and non-intestinal infectious conditions.

Orally ingested probiotic micro-organisms do not exert health effects exclusively in the intestine. Some strains can alleviate or prevent bacterial, fungal or viral infections in other organs by stimulation of the immune system. By preservation or improvement of the barrier function of the intestinal mucosa, they may inhibit translocation of potential pathogens and thus prevent infections of the blood stream and other tissues and organs.

Inulin, oligofructose and mineral metabolism - experimental data and mechanism.

Numerous investigations performed in animal models in the past 10 years have shown repeatedly that non-digestible oligosaccharides (NDO), such as inulin, oligofructose or transgalacto-oligosaccharides (TOS), stimulate mineral absorption, mainly calcium and magnesium. Long-term beneficial effects on bone health have been indicated by accumulation of bone mineral content in growing rats or prevention of bone loss in ovariectomized rats. However, bone mineral content or density are not necessarily associated with bone quality.

Cloning of Escherichia coli lacZ and lacY genes and their expression in Gluconobacter oxydans and Acetobacter liquefaciens.

An efficient transformation protocol for Gluconobacter oxydans and Acetobacter liquefaciens strains was developed by preparation of electrocompetent cells grown on yeast extract-ethanol medium. Plasmid pBBR122 was used as broad-host-range vector to clone the Escherichia coli lacZY genes in G. oxydans and A.

Occurrence and biochemical characteristics of natural bioactive substances in bovine milk lipids.

Bovine milk lipids (BML) contain a number of bioactive substances with positive as well as negative properties, mainly in the class of fatty acids. Besides trans fatty acids (TFA), conjugated linoleic acids (CLA) are of particular interest. Apart from ruminant meat products the main source of CLA in food are BML.

Protein-bound D-amino acids, and to a lesser extent lysinoalanine, decrease true ileal protein digestibility in minipigs as determined with (15)N-labeling.

Heat and alkali treatment of food may increase the concentrations of protein-bound D-amino acids and cross-links such as lysinoalanine (LAL). To examine how protein treatment affects digestibility, purified test meals [total protein 150 g/kg dry matter (DM), 0.44 MJ/(kg BW(0.

Bioactive peptides encrypted in milk proteins: proteolytic activation and thropho-functional properties.

The bioactivities of peptides encrypted in major milk proteins are latent until released and activated by enzymatic proteolysis, e.g. during gastrointestinal digestion or food processing.

Detection of incurred dihydrostreptomycin residues in milk by liquid chromatography and preliminary confirmation methods.

An LC method for the determination of the aminoglycosides streptomycin (STR) and dihydrostreptomycin (DHS) in milk was developed/modified on the basis of published papers. Mean recoveries were 87 and 95% for STR and DHS, respectively. Recoveries are dependent on the concentration level and batch of solid-phase extraction columns used, and independent of fat content and homogenization.

Recovery of 15N-lactoferrin is higher than that of 15N-casein in the small intestine of suckling, but not adult miniature pigs.

Performance of biological functions of lactoferrin in the small intestine requires at least some resistance to degradation. Therefore, we studied prececal digestibility of lactoferrin in comparison to casein both in suckling and adult miniature pigs, applying 15N-labeled proteins. In study 1, 43 piglets (10-d-old), deprived of food for 12 h received 10 mL of sow's milk supplemented with 120 mg of 15N-labeled protein (porcine or bovine lactoferrin or bovine casein).

Crosslinking of sodium caseinate by a microbial transglutaminase.

Sodium caseinate is an effective substrate for transglutaminase. Crosslinking takes place at fast reaction rates resulting in high degrees of crosslinking. The polymers can be hydrolysed by trypsin, but the proteolysates contain residual portions of non- or partlyhydrolysed aggregates.

Evidence for peptide synthesis in the course of in vitro proteolysis.

The present paper reports on studies concerned with furnishing evidence for peptide synthesis in the course of in vitro proteolysis. To this end, the oxidized chain B from insulin (INS) (S = 5% in demineralized water) was subjected to tryptic proteolysis (E/S = 1/50; pH 5; 37 degrees C; 24 h). HPLC-as well as amino acid-and sequence analytical studies have shown that the heptapeptide INS 23-29 (Gly-Phe-Phe-Tyr-Thr-Pro-Lys) liberated by way of hydrolysis is linked by tryptic synthesis via transpeptidation or condensation to form a dimer which accounts for 15% of the amount of monomer.

Biochemical properties of regulatory peptides derived from milk proteins.

Biologically active peptides derived from milk proteins are inactive within the sequence of the precursor proteins but can be released by enzymatic proteolysis. Based on structure-activity studies, peptides with a defined bioactivity show common structural features. Moreover, many milk protein-derived peptides reveal multifunctional bioactivities.

Comparison of Biosensor, Microbiological, Immunochemical, and Physical Methods for Detection of Sulfamethazine Residues in Raw Milk.

A biosensor assay based on biospecific interaction analysis (BIA) was compared with already existing methods for detection of sulphamethazine (SMZ) residues in milk. Microbial inhibitor and receptor assays, an enzyme-linked immunosorbent assay (ELISA), high-pressure liquid chromatography (HPLC), and BIA were used to analyze milk samples from SMZ-treated cows. The results of the commercially available tests (Delvotest SP Special, BR-test Blue Star, Charm II test) were in agreement with the claimed sensitivity of the respective assays.

Detection of inhibitors in milk by microbial tests. A review.

The demands concerning microbial inhibitor tests were subjected to marked changes during the last decades: It started with the claim of being able to detect contaminated milk which might cause problems during fermentation processes ('technological safety'). Due to the present day attention drawn to toxic and allergic hazards for numerous antimicrobials Maximum Residue Limits (MRLs) or safe/tolerance levels are fixed ('toxicological safety'). This means a variety of demands underlying permanent changes with respect to the 'detection pattern' which cannot be fulfilled by a single test.