• 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.

• Applied Biological Chemistry
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• v.4
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• pp.33-42
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• 1963

1. Two fermentation methods have been compared for the production of Takju (Korean native Sake). One method (S1) uses 'Nuruk' (Korean mold wheat) and the other method (S2) uses 'Nuruk' and mold rice. The chemical analysis of source material, also, have been made in order the check the quantitative changes during the fermentation. The results are summarized below: a. The rate of fermentation was slower for the S2 method than for the S1 method. However, the residual sugar in the S2 method was smaller and the amount of alcohol produced the same as with the S1 method. This was true in spite of the fact that the total initial sugar content for the S2 method was 10 percent below that of S1. b. With both the S1 and S2 methods, 80 percent of the total alcohol production was obtained with in 3 to 4 days. c. The pH value of the base material at the beginning of fermentation was markedly different between the two methods I.e.6.0 to 6.2 for S1 and 4.8 to 5.2 for S2. However, after one or two day the pH of both materials was about the same 4.0 to 4.2 and remained at this value unit the fermentation was complete. 2. Organic acids and Sugars in the source materials have been detected by the paper partition chromatography(p.p.c.) method and the followings are obtained. a. Important Organic acids are Fumalic, malic Succinic, Citric, Acetic aicds in polished rice and Fumalic, Succinic, Acetic, Citric, Malic and Oxalic acids 'Nuruk'. The same kinds of acids as in the rice are found in mold rice, However, amount of Citrice acid is markedly increased in mold rice. b. The important Sugar, Glucose, Fructose and Raffinose in polished rice, Gulcose, Fructo, se, Xylose and one which supposed to be Kojibjose in 'Nuruk'. Glucose and those supposed to be Isomaltose, Kojibiose and Sakebiose and found in rice mold, however, no Sucrose, which was exist in polished rice, was found. 3. The important Organic acids found in fermenting mash using the p.p.c. method were Lactic, Succinic and Acetic. Citric acid identified early in the fermentation, S2 method. remained throughout. However, with the S1 method Cirtic acid was detected only during the late stage of fermentation. Sugars not found in the original materials were two which supposed to be Isomaltotriose and Pentose. Maltose found at the beginning of the fermentation disappeared within one day, Isomaltose was detected throughout the period of fermentation. 4. The Somogyi method which was employed to determine the quantitative changes of sugars in the orginal meterial and mach, showed that polished rice containes in order and in largest amounts Sucrose, Glucose, Raffinose and Fructose. 'Nuruk' contained almost equal quantities of Glucose and Fructose. However, the Glucose content of the mold rice exceeded that originally in the polished rice by 25 or 30 times. Only a small quantity of free sugars was found in the mash at the end of the fermentation.