John Sørensen

Phyto-oestrogens in herbage and milk from cows grazing white clover, red clover, lucerne or chicory-rich pastures.

A grazing experiment was carried out to study the concentration of phyto-oestrogens in herbage for cattle and in milk during two periods (May and June). Forty-eight Danish Holstein cows were divided into four groups with four treatment diets; white clover, red clover, lucerne and chicory-rich pastures. Each experimental period lasted 15 days. Herbage samples from the first day and individual milk samples from the last day of the experimental period were analysed for phyto-oestrogens using LC-MS technique. The total concentration of phyto-oestrogens was 21 399 mg/kg dry matter (DM) for red clover and 238 to 466 mg/kg DM for the other three herbages mainly due to a much higher concentration of biochanin A, formononetin and glycitein in red clover. In the milk, the total concentration of phyto-oestrogens was 253 to 397 μg/l for red clover milk and 56 to 91 μg/l in the milk from the other three treatments. This was especially due to a higher concentration of equol, daidzein and formononetin in the red clover milk. The concentration of biochanin A was significantly higher in milk from the red clover treatment in May while no differences were observed in June. Enterodiol was similar across treatments while the concentration of enterolactone was significantly lower for red clover milk compared with the other treatments. Of the tested pastures, red clover appears to have the highest concentration and to be the best source of phyto-oestrogens, especially equol, in bovine milk.

Effect of specific wavelengths on light-induced quality changes in Havarti cheese

The effects of exposure of slices of Havarti cheeses to monochromatic light of wavelengths 366 nm, 405 nm, and 436 nm, respectively, were studied by tristimulus colorimetry, solid-phase microextraction gas chromatographic analysis of volatiles, and open-end fluorescence spectroscopy. Having determined the photon fluxes of the three wavelengths by ferrioxalate actinometry, it was possible to quantify the effects of light exposure in an absolute manner. For all analyses, the most severe effects were caused by visible light, leading to colour bleaching, change in hue, riboflavin degradation, and formation of the secondary oxidation products hexanal, 1-pentanol, and 1-hexanol. Apparent quantum yields for formation of hexanal and 1-pentanol were found to be insignificantly different for 405 nm and 436 nm exposures, having values of (3-5) x 10(-5) mol x einstein(-1) and (9-13) x 10(-5) mol x einstein(-1), respectively. These compounds were not formed when exposed to 366 nm light. In contrast, 1-hexanol was formed when exposing cheese to all three wavelengths, resulting in apparent quantum yields of (2-6) x 10(-5) mol x einstein(-1). The results obtained are discussed in relation to the interplay between inherent product colorants, light sources, and transmission characteristics of the packaging materials.

Comparison of ripening characteristics of Danbo cheeses from two dairies

Abstract Chromatographic and electrophoretic methods have been established as valuable tools in characterising cheese ripening. In a recent IDF-Bulletin, a number of such methods have been described. Some of the methods have been used in the analyses of Danbo 45+ cheese from 41 vats produced in two different dairies. The purpose of this work was to investigate if the two dairies produced cheese with systematic differences in ripening characteristics and thereby possibly also in quality. Casein breakdown was monitored by capillary electrophoresis, peptides by reversed phase liquid chromatography and flavour compounds by gas chromatography-mass spectrometry. Multivariate analysis of the data was performed by principal component analysis, which revealed systematic differences between the dairies, most clearly in caseins and in peptides. Data from chemical and sensory analyses of a subset of the cheeses showed systematic variations in the chemical variables but only minor differences were found in the sensory attributes.

Protease-induced aggregation of bovine α-lactalbumin: Identification of the primary associating fragment

Details in the hydrolysis of alpha-lactalbumin known to result in formation of highly ordered nanotubules, was investigated by incubation of solutions with 10 g alpha-lactalbumin/l with a specific protease from Bacillus licheniformis (BLP). After 50 min of incubation, soluble aggregates were formed, the concentration of which increased until precipitation occurred after 200 min. The latter aggregates were dissolved in urea or at low pH, like the nanotubules characteristic of gels formed by the action of BLP on alpha-lactalbumin at 100 g/l. On the molecular level, alpha-lactalbumin was initially cleaved into two large hydrophobic fragments with masses of 11.6 and 11.3 kDa, which in turn were cleaved in a stepwise manner into the ultimate fragment of 8.8 kDa. This fragment was the predominating component in the insoluble aggregates, and was identified as the sequences 26-37 and 50-113 of alpha-lactalbumin linked together by a disulphide bond. Cleavage of alpha-lactalbumin into this fragment probably created new hydrophobic surfaces and new calcium binding sites allowing its association into ordered structures.

Towards Control of Aggregational Behaviour of α-Lactalbumin at Acidic pH

Abstractα-Lactalbumin (α-La) undergoes considerable structural changes upon loss of bound Ca2+ at acidic pH, leaving α-La in a molten globule structure. Using fluorescence the present work provides more insight into the structural transition of α-La at acidic pH leading to protein aggregation, most likely caused by a combination of hydrophobic and electrostatic interactions. The rate of aggregation is determined by the protein concentration and temperature applied. Availability of Ca2+ stabilises the protein, and thus prevent aggregation at pH values as low as pH 2.9. In contrast, presence of Cu2+ induces a destabilisation of the protein, which can be explained by a binding to the Zn2+ binding site in α-La, possibly resulting in structural alterations of the protein. In general, presence of anions destabilise α-La at pH values below pI, with SO42− exhibiting the strongest effect on the protein stability, thus correlating well with the Hofmeister series. At more acidic pH values far from pI, α-La becomes more stable towards ion induced aggregation, since higher ion activity is required to efficiently screen the charges on the protein surface. The results presented in this paper provide detailed knowledge on the external parameters leading to aggregation of α-La at acidic pH, thus permitting rational design of the aggregation process.

Rapid quantification of casein in skim milk using Fourier transform infrared spectroscopy, enzymatic perturbation, and multiway partial least squares regression: Monitoring chymosin at work

In this study, we introduce enzymatic perturbation combined with Fourier transform infrared (FTIR) spectroscopy as a concept for quantifying casein in subcritical heated skim milk using chemometric multiway analysis. Chymosin is a protease that cleaves specifically caseins. As a result of hydrolysis, all casein proteins clot to form a creamy precipitate, and whey proteins remain in the supernatant. We monitored the cheese-clotting reaction in real time using FTIR and analyzed the resulting evolution profiles to establish calibration models using parallel factor analysis and multiway partial least squares regression. Because we observed casein-specific kinetic changes, the retrieved models were independent of the chemical background matrix and were therefore robust against possible covariance effects. We tested the robustness of the models by spiking the milk solutions with whey, calcium, and cream. This method can be used at different stages in the dairy production chain to ensure the quality of the delivered milk. In particular, the cheese-making industry can benefit from such methods to optimize production control.

Peptidomics in extended shelf life dairy products, for assessment of proteolysis and product quality

Some extended shelf life dairy products develop insoluble material which is referred to as unwanted protein aggregation. This phenomenon appears at varying time points during the shelf life. It occurs seemingly randomly in some productions and not in others for the same product. So far no decisive factor for its development has been identified, indicating protein chemical based differences between the products, due to biological diversities in the applied raw material. This is a very plausible assumption since milk is supplied from multiple farms to the dairies. In this particular project we used re-hydrated industrial whey protein isolate (WPI) produced from cheese whey. This originates from multiple cheese dairies, which also adds to the inhomogeneity of the product.