• KSBB Journal
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• v.14 no.4
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• pp.422-428
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• 1999

A transformat Alcaligence eutrophus GA5 harboring phbC gene from A. latus was cultivated for production of Poly(3-hydroxybutyrate-4-hydroxybutyrate)[P(3HB-4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB). Transformation did not influenced significantly on total cell growth, on total cell growth, concentration, and content of P(3HB-4HB), however, significantly influenced on 4HB molar fraction in P(3HB-4HB) increasing from 12.3 to 23.5 mol% after 48 h cultivation in two-stage using 1.0%(W/V) of ${\gamma}$-butyrolactone as a precursor compare to parent strain. Above increment may be due to the accelerated polymerization between 3HB and 4HB converted from precusor compound by amplified phbC gene. Citrate increased remarkbly total cell mass and P(3HB-4HB) concentration, but did not influenced on the molar fraction of 4HB, meanwhile, magnesium ion influenced on P(3HB-4HB) concentration and 4HB molar fraction significantly. The two-stage cultivation method was modified, in such a way minimizing P(3HB) accumulated inside of cell grown at first-stage, consquently, 26.3% of P(3HB-4HB) containing 61.0 mol% of 4HB fraction was obtained after 72hr. Furthermore, semi-homopolymeric P(4HB) containing 92.0 mol% of 4Hb was obtained, and its structure was confirmed by $^1$H-NMR.

• KSBB Journal
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• v.25 no.3
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• pp.239-243
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• 2010

Six strains of Ralstonia eutropha were grown to investigate characteristics of polyhydroxyalkanoates (PHA) synthesis from vegetable oils or glycerol. Poly(3-hydroxybutyrate) homopolymer was formed using soybean oil, olive oil, or glycerol as carbon source, while poly(3-hydroxybutyrate-co-4-hydroxybutyrate) or poly(3-hydroxybutyrateco-3-hydroxyvalerate) copolymers were synthesized by co-feeding $\gamma$-butyrolactone or pentanoic acid, respectively. Optimum strain was determined as R. eutropha KCTC 2662 in terms of final cell concentration and PHA content. From 20 g/L of soybean oil (optimum substrate), cell concentration and PHA content at 72 h ranged 1.7~9.2 g/L and 70~92 wt%, respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.603-606
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• 2005

Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.

• Proceedings of the PSK Conference
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• 2001.10a
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• pp.282.1-282.1
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• 2001

• BMB Reports
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• v.32 no.3
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• pp.254-258
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• 1999

The NADPH-dependent succinic semialdehyde reductase is one of the key enzymes in the brain GABA shunt, and it catalyzes the formation of the neuromodulator $\gamma$-hydroxybutyrate from succinic semi aldehyde. This enzyme was inactivated by diethylpyrocarbonate (DEP) with the second-order rate constant of $1.1{\times}10^3\;M^{-1}min^{-1}$ at pH 7.0, $25^{\circ}C$, showing a concomitant increase in absorbance at 242 nm due to the formation of N-carbethoxyhistidyl derivatives. Complete inactivation of succinic semialdehyde reductase required the modification of five histidyl residues per molecule of enzyme. However, only one residue was calculated to be essential for enzyme activity by a statistical analysis of the residual enzyme activity. The inactivation of the enzyme by DEP was prevented by preincubation of the enzyme with the coenzyme NADPH but not with the substrate succinic semialdehyde. These results suggest that an essential histidyl residue involved in the catalytic activity is located at or near the coenzyme binding site of the brain succinic semialdehyde reductase.

• Animal Bioscience
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• v.34 no.4
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• pp.575-583
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• 2021

Objective: The aim of the study was to determine the effect of parity and the stage of lactation on the metabolic profile of cows based on the basic chemical milk components and the blood parameters. Methods: The study material consisted of high-yielding Holstein-Friesian cows. In total, 473 cows were examined. According to the parity, cows were divided into four groups: primiparous (P), and multiparous in the second (M2), in the third (M3), and in subsequent lactations (M4). The feeding of cows was based on total mixed ration (TMR) ad libitum. Milk and blood samples were collected individually from each cow three times per standard lactation period. Results: Greater exacerbation of changes in the dynamics of the blood plasma parameters examined was proved for multiparous cows. The highest value of β-hydroxybutyrate acid (0.946 mmol/L) was found for multiparous cows from group M3 at the beginning of lactation. However, it was still in the normal range. The results showed aspartate aminotransferase, and gamma-glutamyl transferase (GGT) activities in dairy cows during lactation had significant variations taking in to account stage of lactation. The highest activity of GGT was found in the group of the oldest cows and measured from 26.36 U/L at the beginning of lactation to 48.75 U/L at the end of the lactation period. Conclusion: The time-related changes in the concentrations of the biochemical parameters described differ markedly among lactating cows, though the housing conditions on the research dairy farm are highly standardised. This indicates that the ability to cope with metabolic stress is mainly affected by the individual predispositions of cows and feed nutrient supply in different stage of lactation. Especially, the feed nutrient supply (in net energy for lactation), which was the best in TMR 1 in comparison TMR 3.

• BMB Reports
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• v.28 no.2
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• pp.112-117
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• 1995

Succinic semialdehyde reductase was inactivated by o-phthalaldehyde. The inactivation followed pseudo-first order kinetics, and the second-order rate constant for the inactivation process was 28 $M^{-1}s^{-1}$ at pH 7.4 and $25^{\circ}C$. The absorption spectrum ($\lambda_{max}$ 337 nm) and fluorescence excitation ($\lambda_{max}$ 340 nm) and fluorescence emission spectra ($\lambda_{max}$ 409 nm) were consistent with the formation of an isoindole derivative in the catalytic site between a cysteine and a lysine residue approximately about 3 $\AA$ apart. The substrate, succinic semialdehyde, did not protect enzymatic activity against inactivation, whereas the coenzyme NADPH protected against o-phthaladehyde induced inactivation of the enzyme. About 1 isoindole group per mol of the enzyme was formed following complete loss of enzymatic activity. These results suggest that the amino acid residues of the enzyme participating in a reaction with o-phthalaldehyde are cysteinyl and lysyl residues at or near the NADPH binding site.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.5
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• pp.648-656
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• 2019

Objective: An experiment was conducted to determine the effect of reduced energy density of close-up diets on metabolites, lipolysis and gluconeogenesis in cows during the transition period. Methods: Thirty-nine Holstein dry cows were blocked and assigned randomly to three groups, fed a high energy density diet (HD, 1.62 Mcal of net energy for lactation $[NE_L]/kg$ dry matter [DM]), a medium energy density diet (MD, $1.47Mcal\;NE_L/kg\;DM$), or a low energy density diet (LD, $1.30Mcal\;NE_L/kg\;DM$) prepartum; they were fed the same lactation diet to 28 days in milk (DIM). All the cows were housed in a free-stall barn and fed ad libitum. Results: The reduced energy density diets decreased the blood insulin concentration and increased nonesterified fatty acids (NEFA) concentration in the prepartum period (p<0.05). They also increased the concentrations of glucose, insulin and glucagon, and decreased the concentrations of NEFA and ${\beta}-hydroxybutyrate$ during the first 2 weeks of lactation (p<0.05). The plasma urea nitrogen concentration of both prepartum and postpartum was not affected by dietary energy density (p>0.05). The dietary energy density had no effect on mRNA abundance of insulin receptors, leptin and peroxisome proliferator-activated $receptor-{\gamma}$ in adipose tissue, and phosphoenolpyruvate carboxykinase, carnitine palmitoyltransferase-1 and peroxisome proliferator-activated $receptor-{\alpha}$ in liver during the transition period (p>0.05). The HD cows had higher mRNA abundance of hormone-sensitive lipase at 3 DIM compared with the MD cows and LD cows (p = 0.001). The mRNA abundance of hepatic pyruvate carboxy-kinase at 3 DIM tended to be increased by the reduced energy density of the close-up diets (p = 0.08). Conclusion: The reduced energy density diet prepartum was effective in controlling adipose tissue mobilization and improving the capacity of hepatic gluconeogenesis postpartum.

• Journal of Veterinary Clinics
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• v.32 no.5
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• pp.426-432
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• 2015

This study compared blood metabolites during peri- and postpartum periods among cows with clinical or subclinical endometritis and cows without endometritis. Blood samples from 207 Holstein dairy cows were collected at 4 weeks prepartum, just after calving, and at 1, 2, 4, and 6 weeks postpartum to measure serum concentrations of calcium, magnesium, non-esterified fatty acids (NEFAs), total cholesterol, albumin, urea nitrogen, ${\beta}$-hydroxybutyrate (BHBA), aspartate aminotransferase (AST), ${\gamma}$-glutamyltransferase, glucose, and phosphorus. Clinical endometritis was diagnosed by the observation of vaginal discharge (> 50% pus) and subclinical endometritis was diagnosed by the evaluation of uterine cytology (> 18% neutrophils) at 4 weeks postpartum. Cows were divided into three groups based on the presence or absence of clinical or subclinical endometritis: the control group (n = 104), the clinical endometritis group (n = 66), and the subclinical endometritis group (n = 37). Calcium and magnesium concentrations were lower in the clinical endometritis group than in the control and subclinical endometritis groups throughout the study period (p < 0.05 to 0.0001), whereas the NEFAs concentration was higher in the clinical endometritis group than in the control group throughout the study period (p < 0.01). The total cholesterol concentration was lower in the clinical endometritis group than in the control and subclinical endometritis groups throughout the pre- and postpartum periods (p < 0.05 to 0.001). The albumin concentration was lower in the clinical endometritis group than in the control and subclinical endometritis groups during the postpartum period (p < 0.05 to 0.001). The urea nitrogen concentration was lower in the clinical endometritis group than in the control and subclinical endometritis groups at 4 and 6 weeks postpartum (p < 0.01). At 1 week postpartum, the BHBA concentration was higher in the clinical endometritis group than in the control group (p < 0.05), whereas the AST concentration was higher in the clinical endometritis and subclinical endometritis groups than in the control group (p < 0.05). In conclusion, lower serum concentrations of calcium, magnesium, total cholesterol, albumin, and urea nitrogen, but higher concentrations of NEFAs, BHBA, and AST during the postpartum period were associated with the incidence of clinical endometritis, indicating the importance of balanced nutrition during the transition period.

• Korean Journal of Veterinary Service
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• v.42 no.2
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• pp.121-126
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• 2019

Metabolic profile test is used to evaluate nutritional imbalance and metabolic disease in dairy cows. The reference intervals of metabolic parameters may change according to nation, region, decades, and maintenance system. Despite the need to be periodically updated for the reference intervals of metabolic parameters, it has rarely been investigated in Korea. Therefore, this aim of study was to provide the reference intervals of metabolic parameters using dairy cows surveyed in Korea during recent years. A metabolic profile test was conducted for 2,976 clinically healthy dairy cows in Korea. Blood samples were collected for the analysis of serum metabolites. This study provided reference intervals of thirteen metabolic parameters (${\beta}$-hydroxybutyrate [${\beta}-HB$], non-esterified fatty acids [NEFA], glucose, total cholesterol [T-COL], total protein, albumin, globulin, blood urea nitrogen [BUN], aspartate aminotransferase [AST], gamma-glutamyl transferase [GGT], calcium, phosphorus, and magnesium). BUN and AST values of the current study were higher than those of previous studies. In the present study, the other metabolic parameters showed low or similar value compared to previous results. Moreover, ${\beta}-HB$, NEFA, T-COL, ALB, BUN, AST, and GGT values were affected by lactation period. This study provided information on the reference intervals of metabolites in healthy dairy cows in Korea. The reference intervals from the present study would be useful in managing and diagnosing disease of dairy cows. However, careful attention should be given in interpreting disease condition for metabolites affected by lactation.