Protease produced from Bacillus sp. FSE-68 was isolated from Meju, a Korean fermented soybean starter, and was purified by a two-consecutive aqueous two-phase system. The change of partition coefficient (K) in the polyethylene glycol (PEG)/potassium phosphate buffer (PPB) aqueous two-phase system was measured at different pHs (6.0- 9.2), PPB concentrations (8-12%), and temperatures (4 and $20^{\circ}C$). As the PPB concentration in the aqueous two-phase system increased, the protease concentration in the top phase (PEG-rich phase) increased, thereby enhancing the partition coefficient. The minimum partition coefficient of the protease was achieved at pH 7.0, whereas that of the total protein was at pH 6.0. The biggest difference in partition coefficients of total protein and protease occurred at pH 6.0. It was interesting to note that the partition coefficient of protease decreased as the temperature increased. The optimum condition of the primary aqueous two-phase extraction of Bacillus sp. FSE-68 was pH 6.0, 14% (w/w) PPB, and 16% (w/w) PEG at $4^{\circ}C$, and the crude enzyme concentration in this system was 50% (w/w). The protease, which was concentrated in the top phase, was further mixed with 15% (w/w) PPB (pH 7.0) in the ratio of 1:1 at $20^{\circ}C$ to elute the bottom phase (PPB-rich phase). Using these steps, the purification fold achieved was 9.2 with a 44.7% yield.
The responses of whole body protein and glucose kinetics and of nitrogen (N) metabolism to non-protein energy intake (NPEI) were determined using an isotope dilution approach and measurement of N balance in three adult male goats. The diets containing 1.0, 1.5 and 2.0 times ME maintenance requirement, with fixed intake of CP (1.5 times maintenance) and percentage of hay (33%), were fed twice daily for each 21 d experimental period. After an adaptation period of 11 d, N balance was determined over 3 d. On day 17, whole body protein synthesis (WBPS) and glucose irreversible loss rate (ILR) were determined during the absorptive state by a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine, [$^2H_4$]tyrosine and [$^{13}C_6$]glucose, with simultaneous measurements of plasma concentrations of metabolites and insulin. Ruminal characteristics were also measured at 6 h after feeding over 3 d. Nitrogen retention tended to increase (p<0.10) with increasing NPEI, although digestible N decreased linearly (p<0.05). Increasing NPEI decreased (p<0.01) ammonia N concentration, but increased acetate (p<0.05) and propionate (p<0.05) concentrations in the rumen. Despite decreased plasma urea N concentration (p<0.01), increased plasma tyrosine concentration (p<0.05), and trends toward increased plasma total amino N (p<0.10) and phenylalanine concentrations (p<0.10) were found in response to increasing NPEI. Increasing NPEI increased ILR of both glucose (p<0.01) and phenylalanine (p<0.05), but did not affect ($p{\geq}0.10$) that of tyrosine. Whole body protein synthesis increased (p<0.05) in response to increasing NPEI, resulting from increased utilization rate for protein synthesis (p<0.05) and unchanged hydroxylation rate of phenylalanine ($p{\geq}0.10$). These results suggest that increasing NPEI may enhance WBPS and glucose turnover at the absorptive state and improve the efficiency of digestible N retention in goats, with possibly decreased ammonia and increased amino acid absorption. In addition, simultaneous increases in WBPS and glucose ILR suggest stimulatory effect of glucose availability on WBPS, especially when sufficient amino acid is supplied.
Li, Cong;Cai, Wentao;Liu, Shuli;Zhou, Chenghao;Cao, Mingyue;Yin, Hongwei;Sun, Dongxiao;Zhang, Shengli;Loor, Juan J.
Asian-Australasian Journal of Animal Sciences
/
v.33
no.11
/
pp.1725-1731
/
2020
Objective: An initial RNA-Sequencing study revealed that UDP-galactose-4-epimerase (GALE) was one of the most promising candidates for milk protein concentration in Chinese Holstein cattle. This enzyme catalyzes the interconversion of UDP-galactose and UDP-glucose, an important step in galactose catabolism. To further validate the genetic effect of GALE on milk protein traits, genetic variations were identified, and genotypes-phenotypes associations were performed. Methods: The entire coding region and the 5'-regulatory region (5'-UTR) of GALE were re-sequenced using pooled DNA of 17 unrelated sires. Association studies for five milk production traits were performed using a mixed linear animal model with a population encompassing 1,027 Chinese Holstein cows. Results: A total of three variants in GALE were identified, including two novel variants (g.2114 A>G and g.2037 G>A) in the 5'-UTR and one previously reported variant (g.3836 G>C) in an intron. All three single nucleotide polymorphisms (SNPs) were associated with milk yield (p<0.0001), fat yield (p = 0.0006 to <0.0001), protein yield (p = 0.0232 to <0.0001) and protein percentage (p<0.0001), while no significant associations were detected between the SNPs and fat percentage. A strong linkage disequilibrium (D' = 0.96 to 1.00) was observed among all three SNPs, and a 5 Kb haplotype block involving three main haplotypes with GAG, AGC, and AGG was formed. The results of haplotype association analyses were consistent with the results of single locus association analysis (p<0.0001). The phenotypic variance ratio above 3.00% was observed for milk protein yield that was explained by SNP-g.3836G >C. Conclusion: Overall, our findings provided new insights into the polymorphic variations in bovine GALE gene and their associations with milk protein concentration. The data indicate their potential uses for marker-assisted breeding or genetic selection schemes.
This study was designed to observe the effects of both control and atherogenic diets on the cholesterol and triglyceride (TG) in serum and liver of adult rats fed diets supplying two levels of dietary fat and two different sources of dietary protein in early life. For the first experimental period, the rats were assigned into the four diet groups: High fat, casein (HC); High fat, gluten (HG); Low fat, casein (LC): Low fat, gluten (LG). Each group was subdivided into control and atherogenic groups for the second experimental period. Cholesterol and TG were determined in serum and liver after 7 hr fasting. The body weight gain was greater in the rats of the casein groups than those of the gluten groups tut not influenced by the level of the dietary fat. The difference in body weight from the quality of dietary protein in the first period was not disappeared even after the second period. After the first period, higher serum cholesterol was observed in the rats fed either casein or high fat diets. With the second experimental diet, rats fed atherogenic diet showed higher serum cholesterol concentration but lower serum TG levels compared to those fed control diet, regardless the diets fed in the first period. Serum cholesterol level of the rats of both groups which had been fed high fat diets in early life was increased compared to those of the low fat diet groups. This effect was more pronounced with the atherogenic diet groups than control groups. However, no differences were found in serum cholesterol levels resulted from the different types of dietary protein fed in the first period. Serum TG concentration was not influenced by the quality of protein and level of fat in the diet but seemed to be mere affected by the amount of carbohyrates in the diet. Liver cholesterol per unit weight was greater in the gluten diet groups than in the casein groups but total cholesterol was higher in casein fed rats. There were no differences in liver TG among the groups.
Objective: An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO), containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods: Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control), 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results: Ruminal pH, total volatile fatty acids (VFA) concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05) compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05) in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP) bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion:The results of the present study suggested that supplementation of MEO may have limited effects on apparent nutrient digestibility, ruminal fermentation and protozoa and HAP bacteria count, blood cells and metabolites.
Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Lee, Dong-Gi;Sarker, Kabita;Lee, Moon-Soon;Xin, Zhanguo;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.130-130
/
2017
Sorghum bicolor is considered as copper-tolerant species. The present study was conducted to understand the copper tolerance mechanism in Sorghum seedling roots. Morphological and effects of Cu on other interacting ions were observed prominently in the roots when the plants were subjected to different concentrations (0, 50, and $100{\mu}M$) of $CuSO_4$. However, the morphological characteristics were reduced by Cu stress, and the most significant growth inhibition was observed in plants treated with the highest concentration of $Cu^{2+}$ ions ($100{\mu}M$). In the proteome analysis, high-throughput two-dimensional polyacrylamide gel electrophoresis coupled with MALDI-TOF-TOF mass spectrometry was performed to explore the molecular responses of Cu-induced sorghum seedling roots. In two-dimensional silver-stained gels, a total of 422 differentially expressed proteins (${\geq}1.5-fold$) were identified using Progenesis SameSpot software. A total of 21 protein spots (${\geq}1.5-fold$) from Cu-induced sorghum roots were analyzed by mass spectrometry. Of the 21 differentially expressed protein spots from Cu-induced sorghum roots, a total of 10 proteins were up-regulated, and 11 proteins were down-regulated. The abundance of the most identified protein species from the roots that function in stress response and metabolism was significantly enhanced, while protein species involved in transcription and regulation were severely reduced. The results obtained from the present study may provide insights into the tolerance mechanism of seedling roots in Sorghum.
Fan, Yang-Kwang;Lin, Yaun-Lung;Chen, Kuen-Jaw;Chiou, Peter Wen-Shyg
Asian-Australasian Journal of Animal Sciences
/
v.15
no.5
/
pp.658-664
/
2002
This study was conducted to determine a proper feeding regime for lactating Holstein cows during the warm season in Taiwan. In Feeding Trial, 21 lactating Holstein cows were randomly allotted into three feeding regimes in a Completely Randomized Design. The feeding regimes were roughage fed ad libitum along with concentrate fed either twice daily (2C) or four times daily (4C), and total mixed ration (TMR) for 8 weeks. No significant differences among the three feeding regimes were found in body weight changes, and intakes in terms of dry matter, crude protein and net energy. For milk yield and 4% FCM yield, 4C were higher than the other two feeding regimes (p<0.05). No significant differences were found in milk compositions or differences noted in the yields of the respective milk components. TMR was worse than 2C and 4C (p<0.05) in milk production efficiencies in terms of dry matter intake per kg milk yield, crude protein intake per kg milk protein yield, as well as dry matter intake, net energy intake and crude protein intake per kg 4% FCM yield. Three ruminally fistulated Holstein dry cows were randomly assigned into the same three feeding regimes in a 3${\times}$3 Latin Square Design. No significant differences were found among the feeding regimes in ruminal pH, ammonia nitrogen, total VFA concentration, molar percentages of VFA, and acetic acid: propionic acid ratio (C$_2$/C$_3$). Taken all together, roughage fed ad libitum and concentrate fed four times daily was the better feeding regime for lactating cows during the warm season in Taiwan.
In order to study the effect of honey concentration on the quality of honeyed ginseng in the process of manufacturing honeyed ginseng, honeyed ginseng was manufactured under the various condition of honey concentration, 40 brix, 50 brix, 60 brix and 70 brix and the approximate composition, the yield and organoleptic test of them were investigated. The results obtained were as follows. 1. The higher honey concentration was used, the yield of honeyed ginseng was increased. 2. The higher honey concentration was used, the amount of total sugar was increased while the amount of crude saponin, crude protein. crude fat, crude fiber and crude ash was decreased. 3. Free sugars of honeyed ginseng consist of rhamnose, xylose, fructose, glucose, sucrose and maltose, 4. The amount of saponin lost in the process of manufacturing honeyed ginseng was 2.5%-5.0%, therefore honey concentration was not so effective on the loss of saponin. 5. According to organoleptic test, H-3 and H-2 treating honey concentration at 60 brix and 50 brix was the best and better of all.
Jeon G. J.;Choy Y. H.;Cho K. H.;Kim M. J.;Kim H. C.;Choi J. G.;Lee C. W.;Hwang J. M.;Kim J. B.
Journal of Embryo Transfer
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v.20
no.3
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pp.239-253
/
2005
Serum concentrations of Hanwoo steers and bulls as possible indicators of beef quality were analyzed to estimate their correlations with carcass traits. Blood samples were taken 2 months and right before shipping to abattoir and at the time of slaughter. And phenotypic correlation coefficients between serum concentrations and carcass traits were estimated. Beef yield index of steers was positively correlated with serum concentrations of total Protein (0.23), albumin (0.26), and calcium (0.31). But it was negatively co..elated with BUN (-0.30). Loin eye area was positively correlated with BUN (0.17) or with globulin (0.16). Back fat thickness was positively correlated with BUN (0.42) and inorganic phosphorus (0.20) being negatively correlated with total protein (-0.23), albumin (-0.33) and calcium (-0.33). Marbling score in the scale of 1 (scarcely marbled) through 9 (extremely marbled) was positively correlated with BUN (0.28) and negatively with IGF-I and calcium concentrations. Phenotype correlation coefficient of loin eye area with total protein concentration in the serum taken from steers right before shipment was estimated to be -0.16 and that with BUN was estimated to be -0.15. Serum concentrations of IGF, glucose, creatinine and on organic phosphorus from steers measured right before shipment were negatively correlated with respective correlation coefficient estimates as -0.21, -0.21, -0.19 and -0.18. Marbling score was negatively co..elated with serum creatinine (-0.16) measured at that time. Beef yield index of steers was positively correlated (0.31) with age adjusted calcium concentration in the serum taken at the time of slaughter. Correlation between body weight and BUN at slaughter was 0.17 At slaughter, loin eye area was negatively correlated with albumin (-0.19) and back fat thickness was also negatively correlated with age adjusted calcium concentration (-0.38). Marbling was negatively correlated with age adjusted calcium concentration(-0.17). Serum concentrations of testosterone, calcium and inorganic phosphorus taken in 2 months before slaughter were negatively but highly correlated with yield index(0.71, 0.67 and -0.71), respectively. Body weight at slaughter was positively was negatively correlated (0.67) with calcium level while dressing percentage was negatively (-0.69) correlated with serum glucose concentration, 2 months prior to slaughter. Correlation coefficients between back fat thickness and cortisol, between back fat thickness and inorganic phosphate were both positive (0.29 and 0.69). Marbling score was negatively correlated with creatinine (-0.81) and positively with BUN (0.87). Body weight loss during shipping was positively correlated with albumin and inorganic phosphate (0.77, 0.83). Yield index of bulls was positively correlated with serum testosterone concentration (0.66). Dressing percentage was positively and highly correlated with globulin (0.73). Back fat thickness of bulls, however, was negatively correlated with testosterone (-0.60). Loin eye area of bull carcasses was positively correlated with testosterone (0.40). Mar-blaine was negatively co..elated with creatinine (-0.55). Yield index of bulls and age adjusted HDLC concentration at slaughter was negatively correlated (-0.71). Dressing percentage of bulls was positively and highly correlated with globulin concentration (0.70). Back fat thickness was also positively correlated with HDLC (0.69) in the serum taken at slaughter. Correlation coefficients between carcass weight and triglyceride, between loin eye are and testosterone and between marbling score and creatinine or glucose were 0.51, -0.91 and -0.58, respectively.
Won, Beom Young;Shin, Ki Young;Ha, Hyun Jee;Yun, Yeo Sang;Kim, Ye Ri;Lee, Hyung Gun
Journal of the Korean Society of Food Science and Nutrition
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v.44
no.6
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pp.882-887
/
2015
This study evaluated the nutritional compositions of dropwort (Oenanthe javanica) extracts depending on the ethanol concentrations. Extractions were performed with hot water, 50% ethanol, 80% ethanol, and 95% ethanol for 4 hours. Changes in yield, as well as total carbohydrate, crude protein, crude fat, total dietary fiber, free sugar, and mineral (Na, Fe, and Ca) contents were investigated. The highest extraction yield of ethanol extracts was 44.67% in 50% ethanol extract of dropwort. Crude protein content reached a maximum of 6.70% while carbohydrate content was highest at 19.6%, in 50% ethanol extract of dropwort. Crude fat content irregularly increased according to ethanol concentration as compared with hot water extract. Total dietary fiber content decreased in ethanol extract, but these changes were not concentration-related. Total sugar contents were highest in hot water and 80% ethanol extracts. Vitamin A content of ethanol extract was higher than that of hot water extract. Mineral (Na, Ca, and Fe) contents were significantly reduced in ethanol extract according to concentration of ethanol, whereas mineral contents were higher in ethanol extract than in hot water extract. Based on this study, ethanol extract of dropwort is more efficient for development of desirable processed foods.
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