• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.17-17
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• 2003

The milk of transgenic animals may provide an attractive vehicle for large-scale production of hEPO. Since glycosylation is cell type specific, recombinant human EPO (rhEPO) produced in different host cells contain different patterns of oligosaccharides, which could affect the biological functions. However, there have been no reports on the characteristics of rhEPO derived from milk of transgenic animals. To address this objective, several transgenic mice by using pWAPhEPO and/or pBC1hEPO expression vector were produced. However, 2 lines of pWAPhEPO founder female mouse died during late gestational day (day 18) before offspring could be obtained. They showed a severe splenomegaly, Unlike those of pWAPhEPO, mammary gland epithelial cells from biopsies of lactating pBC1hEPO transgenic mice had marked immunoreactivity to EPO and any activity was not detected in other tissues. The expression level of rhEPO is about 0.7% of mammary gland cellular total soluble proteins and an amount of 300~500 mg/L rhEPO is secreted into milk. Furthermore, the pBC1hEPO transgenic mice transmitted this character to their progeny in mendelian manner. In order to determine the extent of glycosylation variation, N-linked oligosaccharide structures present in the milk-derived rhEPO were characterized. Most of milk-derived rhEPO is fully glycosylated. the biological activity of milk-derived rhEPO was comparable to that of purified CHO-derived rhEPO, and milk-derived rhEPO showed relatively stable after freezing and thawing. Taken together, the results illustrate the potential of transgenic animals in the large-scale production of biopharmaceuticals.

• Food Science of Animal Resources
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• v.36 no.4
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• pp.516-522
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• 2016

Goat milk is highly nutritious and is consumed in many countries, but the development of functional foods from goat milk has been slow compared to that for other types of milk. The aim of this study was to develop a goat milk protein hydrolysate (GMPH) with enhanced digestibility and better hypoallergenic properties in comparison with other protein sources such as ovalbumin and soy protein. Goat milk protein was digested with four commercial food-grade proteases (separately) under various conditions to achieve the best hydrolysis of α_s -casein and β-lactoglobulin. It was shown that treatment with alcalase (0.4%, 60℃ for 30 min) effectively degraded these two proteins, as determined by SDS-PAGE, measurement of nonprotein nitrogen content, and reverse-phase high-performance liquid chromatography. Hydrolysis with alcalase resulted in a significant decrease in β-lactoglobulin concentration (almost to nil) and a ~40% reduction in the level of α_s-casein. Quantification of histamine and TNF-α released from HMC-1 cells (human mast cell line) showed that the GMPH did not induce an allergic response when compared to the control. Hence, the GMPH may be useful for development of novel foods for infants, the elderly, and convalescent patients, to replace cow milk.

• Food Science of Animal Resources
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• v.31 no.6
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• pp.851-857
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• 2011

This study was conducted to develop a real time reverse transcriptase-PCR (RT-PCR) method for the detection of viable Enterobacteriaceae in milk using primers based on the genes of ribosomal proteins S11 and S13 and to determine effects of heating and subsequent treatments on the threshold cycle (Ct) of the real time RT-PCR. Total RNA was isolated from 17 strains of bacteria including 11 strains of Enterobacteriaceae suspended in milk using a modified Tri reagent method. SYBR Green Master Mix was added to the RNA and the mixture was subjected to the real time RT-PCR. The Cts of eleven type strains of the Enterobacteriaceae in milk ($10^7$ cells) in the real time RT-PCR ranged from 21.5 to 24.6. However, the Cts of Pseudomonas fluorescens, Acinetobacter calcoaceticus, and three gram-positive bacteria were more than 40. The real time RT-PCR detected as low as $10^3$ cells in agarose gel electrophoresis. The Cts increased from 22.0 to 34.2 when milk samples contaminated with Escherichia coli ($10^7$ cells/mL) were heated at $65^{\circ}C$ for 30 min. In addition, subsequent incubation at $37^{\circ}C$ for 6 and 24 h increased the Cts further up to 36.2 and 37.2, respectively. Addition of RNase A to the bacterial suspension obtained from the heated milk and subsequent incubation at $37^{\circ}C$ for 1 h increased the Cts to more than 40. The results of this study suggests that pretreatment of bacterial cells heated in milk with RNase A before RNA extraction might enhance the ability to differentiate between viable and dead bacteria using real time RT-PCR.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.4
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• pp.564-568
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• 2004

Restructuring had made it possible to utilize lower value cuts and meat trimmings from spent animals by providing convenience in product preparation besides enhancing tenderness, palatability and value. Milk co-precipitates (MCP) have been reported to improve the nutritional and functional properties of certain meat products. This study was undertaken to evaluate the influence of incorporation of milk co-precipitates at four different levels viz. 0, 10, 15 and 20% on the quality of restructured buffalo meat blocks. Low-calcium milk co-precipitates were prepared from skim milk by heat and salt coagulation of milk proteins. Meat chunks were mixed with the curing ingredients and chilled water in a Hobart mixer for 5 minutes, followed by addition of milk co-precipitates along with condiments and spice mix and again mixed for 5 minutes. Treated chunks were stuffed in aluminium moulds and cooked in steam without pressure for 1.5 h. After cooking, treated meat blocks were compared for different physico-chemical and sensory attributes. Meat blocks incorporated with 10% MCP were significantly better (p<0.05) than those incorporated with 0, 15 and 20% MCP in cooking yield, percent shrinkage and moisture retention. Sensory scores were also marginally higher for meat blocks incorporated with 10% MCP than product incorporated with 15 and 20% MCP, besides being significantly higher than control. On the basis of above results 10% MCP was considered optimum for the preparation of restructured buffalo meat blocks. Instrumental texture profile analysis revealed that meat blocks incorporated with 10% MCP were significantly better (p<0.05) in hardness/ firmness than control although, no significant (p>0.05) differences were observed in cohesiveness, springiness, gumminess and chewiness of both type of samples.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.3
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• pp.311-317
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• 1998

A total of 1033 Brown Swiss and 610 Canadienne cows were phenotyped for the genetic variants ${\alpha}_{s1}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\beta}$-lactoglobulin and ${\alpha}$-lactalbumin. In Brown Swiss, frequency distributions were: 97.3% B and 2.7% C variant of ${\alpha}_{s1}$-casein; 31.6% $A^1$, 51.8% $A^2$, 0.5% $A^3$ and 16.1% B variant of ${\beta}$-casein; 70.4% A, 29.3% B, and 0.3% C variant of ${\kappa}$-casein; 41.7% A and 58.3% B variant of ${\beta}$-lactoglobulin; and 100% B variant of ${\alpha}$-lactalbumin. Corresponding frequencies in Canadienne for those five milk proteins were: 98.6 and 1.4%;58.5, 33.5, 0.08 and 7.9%; 78.8, 21.1 and 0.1%, 42.4 and 57.6%; and 100%. Analysis of variance by least squares showed possible association between milk protein phenotypes and some lactational production traits. There were no significant association of phenotypes of ${\alpha}_{s1}$-casein, ${\beta}$-casein and ${\beta}$-lactoglobulin with milk yield, fat yield, protein yield, fat percentage and protein percentage in both breeds during the three lactations. In the Brown Swiss, ${\kappa}$-casein phenotype was associated with 305-day fat yield and protein yield during the first lactation. ${\kappa}$-Casein AB was associated with higher milk, fat and protein yield during the second lactation. During the third lactation, ${\beta}$-lactoglobulin AA in Canadienne cows was associated with higher protein content in the milk (3.70%) when compared to phenotypes AB (3.54%) and BB (3.64%).

• Journal of Nutrition and Health
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• v.22 no.2
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• pp.84-90
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• 1989

Allergic disorders may be defined as the abnormal reactions to proteins. Cow milk is the first foreign protein which the human being would contact postnatally. Serveral studies suggested that this contact had certain effects on the development o allergic disorders with great controlversies. Seven hundred seventy eight students from middle class in Seoul, aged 6 to 17 wears, were surveyed to delineate the relationshpis between the cow's milk contacts in infancy and the subsequent development of allergic disorders in Koran population. Questionaire were included the nursing patterns in infancy, the past medical history of asthma or wheezy bronchitis and the presence of infantile eczema. Skin tests to three common resporatory allergens in Korea(house dust, house dust mite; D. farinae, D. ptoeronssinus) were performed. The positivity was determined by the wheal size larger than that to histamine. 70.2% of whole mateials was fed by breast milk, 13.1% by cow's milk and 16.7% by mixed nutsing. From the obsevation of nursing patterns by (table 2), we find that the number of breast milk group has been decreasing in 1970's. 10.1% of breast milk group(BMG) had wheezy bronchitis, 8.8% of cow's milk group(CMG) and 11.5% of mixed nursing group(MNG). Infantile eczema history was positive in 27.8% of BMG, in 26.5% of CMG and in 29% of MNG. Skin test positivity was 23.8% in BMG, 25.5% in CMG and 26.2% in MNG. There prevalences of allergic disorders in each nursing group did not reveal any statistically significant differences. In conclusion, nursing patterns in infancy do not seem to have certain effects on the development of allergic disorders such as infantile eczema. wheezv bronchitis or skin test positivity.

• Korean Journal of Food Science and Technology
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• v.38 no.2
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• pp.304-308
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• 2006

This study was performed to understand the behavior of protein mobility and intensity of enzymatic hydrolysis according to crosslinking of sodium caseinate, whey protein isolate, skim milk and whole milk powders with or without transglutaminase (TGase, w/w = 200 : 1) at $38^{\circ}C$. Whey protein was limited to crosslinking and skim milk was relatively more increased in high molecular polymer than whole milk. The degree of crosslinking decreased in the order of sodium caseinate>skim milk>whole milk>whey protein isolate. The SDS-PAGE results indicated that main bands of TGase treated samples had a high mobility and formed bands of molecular weights below 15 kDa by hydrolysis with pepsin after 10 min of reaction time. However, ${\beta}-lactoglobulin$ showed remarkable stability against pepsin hydrolysis treated with and without TGase. The high molecular polymers were easily hydrolyzed with digestive enzymes in vitro experiment. These results suggested that novel dairy products using TGase would have no special digestive problem in human body.

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.4
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• pp.381-386
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• 1985

This experiment was conducted to investigate changes of soybean curd yield according to the extension of soaking time during manufacturing of soybean curd. To investigate those changes systematically, transmission electron microscopy and disc-gel electrophoresis were used. The soybean curd yield was increased from 45.0% to 50.5% and 55.4% respectively as soaking time is extended from 5 hours to 10 and 24 hours. The solid extraction and soybean milk coagulation were also increased according to the extension of soaking time. From disc-gel electrophoresis patterns of soybean milk protein and soybean curd protein, numbers of band were increased and major band thickened by expending the soaking time. Most of high molecular bands of soybean milk protein were transfered to soybean curd. Crude 7S proteins of soybean milk and soybean curd in dis-gel electrophoresis were appeared to be 4 and 5 bands respectively, and crude 11S proteins of soybean milk and soybean curd were appeared to be 9 and 8 bands respectively. Of soybean milk bands, most of 11S component transfered to soybean curd. Transmission electron photomicrographs revealed that the dimension of each protein body became larger and the numbers of spherosome around the protein bodies in unit area fewer by extending the soaking time of soybean.

• Journal of Animal Science and Technology
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• v.64 no.3
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• pp.481-499
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• 2022

This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/ F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.

• Near Infrared Analysis
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• v.2 no.1
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• pp.59-66
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• 2001

Nowadays, medical diagnostics is efficiently supported by clinical chemistry and near infrared spectroscopy is becoming a new dimension, which has shown high potential to provide valuable information for diagnosis. The investigation was carried out to study the influence of mammary gland inflammation, called mastitis, on cow´s milk spectra and milk composition measured by near infrared spectroscopy (NIRS). Milk somatic cell counts (SCC) in milk were used as a measure of mammary gland inflammation. Naturally occurred variations with milk composition within lactation and in the process of milking were included in the experimental design of this study. Time series of unhomogenized, raw milk spectral data were collected from 3 cow along morning and evening milking, for 5 consecutive months, within their second lactation. In the time of the trial, the investigated cows had periods with mammary gland inflammation. Transmittance spectra of 258 milk samples were obtained by NIRSystem 6500 spectrophotometer in 1100-2400 nm region. Calibration equations for the examined milk components were developed by PLS regression using 3 different sets of samples: samples with low somatic cell count (SCC), samples with high SCC and combined data set. The NIR calibration and prediction of individual cow´s milk fat, protein, and lactose were highly influenced by the presence of mil samples from animals with mammary gland inflammation in the data set. The best accuracy of prediction (i.e. the lower SEP and the higher correlation coefficient) for fat, protein and lactose was obtained for equations, developed when using only “healthy” samples, with low SCC. The standard error of prediction increased and correlation coefficient decreased significantly when equations for low SCC milk were used to predict examined components in “mastitis” samples with high SCC, and vice versa. Combined data set that included samples from healthy and mastitis animals could be used to build up regression models for screening. Further use of separate model for healthy samples improved milk composition measurement. Regression vectors for NIR mild protein measurement obtained for “healthy” and “mastitic” group were compared and revealed differences in 1390-1450 nm, 1500-1740 nm and 1900-2200 nm regions and thus illustrated post-secretory breakdown of milk proteins by hydrolytic enzymes that occurred with mastitis. For the first time it has been found that monitoring the spectral differences in water bands at 1440 nm and 1912 nm could provide valuable information for inflammation diagnosis.