Glutamic acid uptake in Streptococcus salivarius subsp. thermophilus was energy dependent, the source of energy and adaptation to sugar being important to efficiency of uptake. The disaccharides, lactose and sucrose, stimulated uptake, but cells grown in glucose were more active. Optimum temperature was ˜ 40 °C and pH ˜ 7·0. NaCl was strongly inhibitory to the uptake of glutamic acid although not to that of isoleucine. High specificity existed because only L-aspartic acid was inhibitory.

Gels varying in protein content and composition were prepared from whey proteins by a standard method and their compression and impact strengths were determined. No surface friction could be observed between the gels and the compression plates, and the Poisson's ratio was close to 0·5. Thus, the elastic moduli of the gels were determined. Compression and impact strengths and elastic moduli were related logarithmically to the β-lactoglobulin content of the protein powders. The strength and manner of fracture of the gels depended on the gel microstructure. Strong gels had a honeycomb structure consisting of a very tightly bound network, and failed by cracking on impact and by shear along cleavage planes in compression. Weaker gels had a loose open network with no definite means of fracture.

The effect of Ca2+. binding and ionic strength on the physicochemical characteristics of phosphoserine residues was studied on O-phospho-DL-serine, bovine β-casein and its phosphopeptide (1–25) using 31P NMR. pK in various experimental conditions were determined.

The pK of the phosphoserine residues of β-casein and its phosphopeptide (1–25) respectively ranged from 6·46 to 7·21 and from 6·57 to 7·10. pK of O-phospho-DL-serine, β-casein and its phosphopeptide decreased when Ca2+ was bound to the phosphoserine residue or when ionic strength was increased. The binding of Ca2+ to the phosphopeptide (1–25) took place at first on phosphoserine residues 17, 18, 19 which had the highest pK, then, when these were saturated, on residue 15 whose pK was the lowest. When the four sites had bound Ca2+, peaks corresponding to a different complex form appeared in the spectrum.

Streptococcus salivarius subsp. thermophilus (Str. thermophilus) has no group-specific antigen. HC1 extracts of 551 strains of streptococci isolated from cheese or yogurt (123 Str. thermophilus and 428 Enterococcus faecalis – Ent. faecium spp.) were examined with three type-specific antisera prepared against representatives of Str. thermophilus and also with Lancefield group D antisera. All typical (80) or atypical (43) strains of Str. thermophilus reacted with at least one type-specific antiserum and none of the enterococci reacted. Only two strains reacted with both type-specific and group antisera.

Micellar calcium phosphate has the chemical composition and physicochemical properties that are consistent with it being a complex of the phosphate centres of casein with an acidic amorphous calcium phosphate. Similar acidic amorphous calcium phosphates have been prepared in the laboratory and for these, as well as for micellar calcium phosphate, the most appropriate crystalline model compound from which the short-range structure may be derived is brushite, CaHPO4.2H2O. The predicted secondary structures around sites of phosphorylation in the Ca2+-sensitive caseins often comprise an α-hclix-loop-α-helix motif with the sites of phosphorylation in the loop region. This motif may be important in linking the colloidal calcium phosphate with casein in native casein micelles.

Accelerated ripening was successfully achieved in Feta cheese by the addition of a heat-shocked culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, or neutral or acid microbial proteinase. Optimum levels of microbial proteinases were established at which trichloroacetic acid-soluble N (TCA-N) compounds were increased by about 35% in 40 d. Treated cheeses of this age had flavour intensities and body and texture qualities typical of 80 d-old control cheeses. Any further increase in TCA-N by adding larger amounts of enzyme resulted in bitterness. Heat-shocked culture treated cheeses developed similar characteristics to those prepared with neutral or acid proteinases. In contrast to the control cheese, it was necessary in the experimental cheeses to add minute amounts of lamb lipase (0·1–0·2 g/100 kg milk) to the cheese milk to keep the flavour balance. Best organoleptic characteristics were obtained from cheese treated with heat-shocked culture, followed by those treated with neutral or acid proteinase.

Enhanced stability of β-galactosidase (Streptococcus salivarius subsp. thermophilus) in milk is due to several milk components acting in concert. Milk salts stabilized the enzyme 3-fold compared to phosphate buffer. K+ was a better stabilizer than Na+. Omission of divalent cations, especially Mg2+, caused a marked drop in stability. Lactose stabilized enzyme stored in the frozen state but destabilized enzyme stored unfrozen. Caseinate stabilized the enzyme 8-fold in phosphate buffer but it stabilized 144-fold in the presence of lactose. In the presence of milk proteins, lactose was 25 times as effective as galactose and 100 times as effective as glucose at promoting stability; sucrose slightly destabilized the enzyme. Stability rose with increasing lactose concentration but declined with increasing enzyme concentration. In milk, soluble casein was the primary stabilizer; whey proteins and peptides had much less effect. Micellar casein had no effect on stability.

β-Galactosidase from an autolytic strain of Streptococcus salivarius subsp. thermophilus was purified 109-fold to near homogeneity. The yield of purified enzyme was 41% and the specific activity was 592 0-nitrophenyl β-D-galactopyranoside U/mg at 37 °C. Two isozymes were present, but only one subunit was detected, having a mol. wt of 116000. Enzyme stability was 37–83 times greater in milk than in buffer in the range 60–65 °C. At 60 °C the half-life in milk was 146 min. Denaturation in buffer was first-order, but in milk the overall reaction order with respect to enzyme concentration was ˜ 0·5. The activation energy for denaturation was 453 kJ/mol in milk and 372 kJ/mol in buffer. In milk the activation energy for lactose hydrolysis was 35·1 kJ/mol.

Milk protein genetic polymorphism has a major influence on the composition of milk, and on its processing properties, including yield of cheese (see Schaar et al. 1985; McLean et al. 1984, 1987; McLean, 1987). However, there appears to be little information on the effects of milk protein genetic variants on syneresis of cheese curd. The effect of casein composition on syneresis was studied by Pearse et al. (1986), who found that syneresis was affected only by the level of β-casein. Syneresis is an essential requirement in cheese making from renneted or acidified milk, but is undesirable during the storage of products such as yogurt. Milk for yogurt manufacture is preheated to minimize syneresis and to give maximal firmness of the yogurt coagulum (Tamime & Deeth, 1980). Pearse et al. (1985) showed that the reduction of one-third in the extent of syneresis caused by heating artificial micelle milk (AMM) containing βlactoglobulin (β-lg) in natural concentrations was due to sulphydryl-mediated complex formation between β-lg and micellar κ-casein which appeared to interfere with the micelle–micelle interactions responsible for syneresis. The results presented here were part of a study which investigated the effects of κcasein and κ-lg genetic variants and concentrations on syneresis of curd formed from renneted heated AMM.

Sandwich enzyme immunoassay of Mucor miehei proteinase (from the commercial preparation Fromase) has been developed using horseradish peroxidase as a marker. The enzyme may be determined to a detection limit of 50 ng/ml. Slight cross reactions by chymosin, bovine, porcine and chicken pepsin did not affect the assay. Recovery of Fromase added to cheese extracts varied between 92 and 106%. The method was used for the determination of M. miehei proteinase in different cheese extracts.

Two l·9 1 Upflow Anaerobic Sludge Blanket (USAB) reactors, operated at 35 °C, were used for the digestion of deproteinated cheese whey substrate. The C/N ratio was 19, and the alkalinity requirements of the two reactors were satisfied by adding Ca(OH)2 and Na2CO3 in two different ratios respectively. Gas and effluent parameters indicated that reactor 1 (receiving a higher level of calcium) was superior and eventually a loading rate of 7·1 kg COD m−3 d−1 could be obtained in this reactor, compared to a loading rate of 5 kg COD m−3 d−1 which was obtained in reactor 2. Pellets with good settling abilities formed in reactor 1, while the sludge in reactor 2 developed a granular slimy appearance and settled poorly. Investigation of the ultrastructure of the sludge particles revealed a difference in the types of bacteria present in the granules.

The influence of pH, temperature and Ca depletion on bovine casein micelle suspensions in D2O containing simulated milk ultrafiltrate was studied by 1H-NMR spectroscopy.

In the pH range of 5·8–7·5 the spectrum of the micelles showed very little pH dependence, indicating that no changes occurred in the dynamic behaviour of the proteins constituting the micelle.

The NMR spectrum of casein micelles was strongly temperature dependent, particularly in the temperature range of 60–98 °C. Increase in temperature resulted in a strong increase in spectral intensity concomitant with changes in the spectral characteristics. In micelle suspensions these changes were reversible, and indicated that at clevated temperatures the rigid structure of the casein micelle started to melt, leading to an increased mobility of appreciable parts of the proteins in the micelle.

Ca depletion of the casein micelles by addition of EDTA resulted in an increase in spectral intensity, which arose from the presence of casein components in the serum phase, The spectrum of these serum phase particlcs resembled closely the spectrum of a solution of total casein in simulated milk ultrafiltrate and was quite different from the spectrum of casein micelles.

The implications of these results with respect to models of the structure of bovine casein micelles are discussed.

The extent and level of proteolysis were evaluated in Cabrales and five other European blue vein cheese varieties, Bleu de Bresse, Danablu, Edelpilzkäse, Gorgonzola and Roquefort, by analysing their different N-containing constituents and determining the degree of casein hydrolysis. The electrophoretic patterns of the caseins in the cheeses confirmed their strong proteolysis, components featuring the lowest electrophoretic mobility offering the greatest resistance to subsequent hydrolysis. Component αs1 — I was detected in all varieties studied with the exception of Roquefort cheese.

The association of non-phosphorylated (0-P) and fully phosphorylated (5-P) human β-caseins was studied by fluorescence spectroscopy and laser light scattering. The tryptophan fluorescence intensity (FI) level increased between 20 and 35 °C, indicating a change in the environment of that residue. A similar transition occurred when ANS was used as a probe. Transition temperatures were slightly lower in 10 mM-CaCl2 but were not affected by an equivalent increase in ionic strength caused by NaCl. The magnitude of the FI change was less for the 5-P than the 0-P protein but was increased for both by CaCl2 addition. These FI data were characteristic of a conformational change and this was supported by fluorescence polarization which indicated that with CaCl2, tryptophan and ANS mobility increased at the transition temperature even though the extent of protein association also increased. Light scattering suggested that protein association proeeeded with the primary formation of submicellar aggregates containing 20–30 monomers which then associated further to form particles of minimum micelle size (12–15 submicelles), and eventually larger. The temperature of precipitation of the 5-P form in the presence of CaCl2 was lower than the conformational transition and suggested that both hydrophobic interactions and Ca bridges between phosphate esters on adjacent molecules are important in micelle formation.

Milk fat-coated microcapsules containing Brevibacterium linens and methionine were used to produce methanethiol, one of the volatile sulphur compounds implicated in Cheddar cheese flavour. Production of methanethiol from methionine occurred aerobically and anaerobically, but the production was 3- to 4-fold greater aerobically with most of the methanethiol being oxidized to dimethyl disulphide. About 35% of the total methanethiol was absorbed by the milk fat capsules and about 65% detected in the headspace. Levels of methanethiol began to decline at 26 °C after 24 h in milk fat-coated microcapsules. However, low temperature, such as 4–12 °C, stabilized levels of methanethiol in microcapsules over an 8-d analysis period. Optimum pH and temperature for methanethiol production were 8 and 26 °C respectively. The antioxidants butylated hydroxytoluene, butylated hydroxyanisole and ascorbic acid had negligible effects on methanethiol production.