Characterization of Bioactive Recombinant Human Lysozyme Expressed in Milk of Cloned Transgenic Cattle

BACKGROUND: There is great potential for using transgenic technology to improve the quality of cow milk and to produce biopharmaceuticals within the mammary gland. Lysozyme, a bactericidal protein that protects human infants from microbial infections, is highly expressed in human milk but is found in only trace amounts in cow milk. METHODOLOGY/PRINCIPAL FINDINGS: We have produced 17 healthy cloned cattle expressing recombinant human lysozyme using somatic cell nuclear transfer. In this study, we just focus on four transgenic cattle which were natural lactation. The expression level of the recombinant lysozyme was up to 25.96 mg/L, as measured by radioimmunoassay. Purified recombinant human lysozyme showed the same physicochemical properties, such as molecular mass and bacterial lysis, as its natural counterpart. Moreover, both recombinant and natural lysozyme had similar conditions for reactivity as well as for pH and temperature stability during in vitro simulations. The gross composition of transgenic and non-transgenic milk, including levels of lactose, total protein, total fat, and total solids were not found significant differences. CONCLUSIONS/SIGNIFICANCE: Thus, our study not only describes transgenic cattle whose milk offers the similar nutritional benefits as human milk but also reports techniques that could be further refined for production of active human lysozyme on a large scale

Bioactive peptides in dairy products.

Bioactive peptides are hydrolysates with specific amino acid sequences that exert a positive physiological influence on the body. They are inert within the native protein, but once cleaved from the native protein by microbial or added enzymes and/or gastrointestinal enzymes during the digestive process, they apply their beneficial traits. Dairy products, particularly fermented products, are potential sources of bioactive peptides: several of them possess extra-nutritional physiological functions that qualify them to be classified under the ‘Functional Foods’ label. Biological peptides in milk, the methods of their generation and their prevalence in dairy products are reviewed along with the reported health benefits and safety aspects

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Not AvailableWhey is a nutritious by product of some traditional Indian processed milk products and it needs to be utilized in an effective way in order to reduce environmental hazards associated with its untreated disposal. Low calorie watermelon beverage appears to be a simple, attractive and economic method of whey disposal. The experiment was designed by Central Composite Rotatable Design of Responce Surface Methodology. Three independent variables whey, Innova ® fiber and sucralose were chosen at five levels within the respective ranges of 40–60 %, 2.0–5.0 % and 0.01–0.03 %. The effect of the variables on flavour, mouthfeel, after-taste, viscosity, total soluble solids (all to be maximized) and sedimentation (to be minimized) was observed. These three were the independent variables whose effect on flavour, mouthfeel, after-taste, viscosity, total soluble solids (all to be maximized) and sedimentation (to be minimized) were evaluated. Quadratic model fitted well to all dependent variables. The R2 values for flavour, mouthfeel, aftertaste, viscosity, sedimentation and TSS were 95.57, 98.71, 95.50, 97.87, 99.26 and 98.17 %, respectively. Response surface methodology was used to optimize the level of processing parameters. Maximum scores for flavour (7.46), mouthfeel (7.49), after-taste (7.72), viscosity (13.55 cp) and total soluble solid (15.34°Brix) and minimum score for sedimentation (1.55 ml/10 ml) were obtained when the formulation contained 51.46 % whey, 3.84 % Innova® fiber and 0.021 % sucralose.Not Availabl

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Not AvailableWatermelon juice was exposed to the enzyme masazyme at varying enzyme concentrations (0.01–0.1 % w/w) and different time (20–120 min) and temperature (30–50 °C) combinations. The effects of the treatments on selected responses (juice recovery, total dissolved solids (TDS), viscosity, turbidity, cloud stability and L value) were determined employing a second order Box Behnken Design in combination with Response Surface Methodology. Enzymatic treatment effectively degraded polysaccharides, resulting in reduced viscosity, turbidity and absorbance value and increased juice recovery, total dissolved solids and lightness. R2 value for all models for the dependent variables were greater than 90 %. The maximum juice recovery (86.27 %), TDS (8.7°Brix) and L value (17.57) while minimum viscosity (0.0020 Pa.s.), turbidity (39.37 NTU) and cloud stability (0.033 abs) were obtained when enzyme treatment was set up with 0.09 % w/w enzyme concentration at 46.90 °C and 117.45 min.Not Availabl

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Not AvailableThe present investigation was carried out to evaluate the immunoregulatory potential of Lactobacillus helveticus fermented milk and its cell free supernatant in mice. Allergic conditions were induced in mice by injecting ovalbumin intraperitoneally on day zero and 14. The mice were divided into four groups and fed milk, L. helveticus fermented milk and its cell free supernatant for a period of 21 days and compared with an allergic control group, respectively. It was observed that on feeding fermented milk and cell free supernatant the production of total IgE and ovalbumin-specific IgE in serum declined significantly in comparison to allergic as well as milk controls. The splenocytes were cultured and cytokines were estimated by ELISA kits. It was observed that levels of Th1 (T helper Type1) cytokines increased whereas the levels of Th2 (T helper Type2) cytokines decreased. Thus decline in IgE production was associated with increase in production of Th1 (IFN-Y, IL-2) cytokines and decrease in Th2 (IL-4, IL-6) cytokines.Not Availabl

Genetic variability of milk fatty acids.

The milk fatty acid (FA) profile is far from the optimal fat composition in regards to human health. The natural sources of variation, such as feeding or genetics, could be used to increase the concentrations of unsaturated fatty acids. The impact of feeding is well described. However, genetic effects on the milk FA composition begin to be extensively studied. This paper summarizes the available information about the genetic variability of FAs. The greatest breed differences in FA composition are observed between Holstein and Jersey milk. Milk fat of the latter breed contains higher concentrations of saturated FAs, especially short-chain FAs. The variation of the delta-9 desaturase activity estimated from specific FA ratios could explain partly these breed differences. The choice of a specific breed seems to be a possibility to improve the nutritional quality of milk fat. Generally, the proportions of FAs in milk are more heritable than the proportions of these same FAs in fat. Heritability estimates range from 0.00 to 0.54. The presence of some single nucleotide polymorphisms could explain partly the observed individual genetic variability. The polymorphisms detected on SCD1 and DGAT1 genes influence the milk FA composition. The SCD1 V allele increases the unsaturation of C16 and C18. The DGAT1 A allele is related to the unsaturation of C18. So, a combination of the molecular and quantitative approaches should be used to develop tools helping farmers in the selection of their animals to improve the nutritional quality of the produced milk fat