• Asian-Australasian Journal of Animal Sciences
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• v.14 no.5
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• pp.615-623
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• 2001

A study of mammary function in relation to glucose metabolism of first lactation Bali cows on grass-legume diets was carried out using 12 primiparous cows (initial BW $263.79{\pm}21.66kg$) for 16 weeks starting immediately post calving. The animals were randomly allocated into 4 dietary treatment groups R1, R2, R3 and R4, receiving from the last 2 months of pregnancy onwards, rations based on a mixture of locally available grass and legume feed ad libitum. On a DM basis R1 contained 70% elephant grass (PP, Penicetum purpureum) plus 30% Gliricidia sepia leaves (GS), R2 was 30% PP plus 25% GS supplemented with 55% Hibiscus tilliacius leaves (HT, defaunating effect), R3 and R4 were 22.5% PP+41.25% GS+11.25% HT+25% concentrate, with R4 supplemented with zinc-diacetate. TDN, CP and zinc contents of the diets were 58.2%, 12.05% and 18.3 mg/kg respectively for R1, 65.05%, 16.9% and 25.6 mg/kg respectively for R2, 66.03%, 16.71% and 29.02 mg/kg respectively for R3 and 66.03%, 16.71% and 60.47 mg/kg respectively for R4. Milk production and body weights were monitored, an energy and protein balance trial conducted, overall glucose kinetics parameters assessed, mammary blood flow (MBF) and metabolite arteriovenous differences (${\Delta}AVs$) measured to get uptake data and mammary performance relationships. Parameters of glucose kinetics at peak lactation or during dry condition were not affected by ration quality. Glucose pool size, space of distribution and flux increased by 61.77, 62.26 and 82.08%, respectively, during lactation compared to the dry period. Mean glucose flux of lactating Bali cows was $5.52mg/min.kgBW^{0.807}$ which resembles the range of values of temperate dairy cows. Calculation showed that glucose requirements for maintenance, milk lactose and fat-glycerol synthesis, and the formation of NADPH reached 461.69 g for a yield of 1 kg/d or equal to 320.62 mg/min, which was less than the average glucose flux of lactating Bali cows of 481.35 mg/min. Mammary blood flow (MBF) values ranged from 56 to 83 l/h for the different treatments and the ratio MBF per kg milk produced improved from av. 1540 l/kg for R1 to av. 967 l/kg for R4 treated cows. Mammary glucose uptake ranged from 6.27 to 12.03 g/h or 120 to 140 g/kg milk. Glucose uptake was mass-wise 2 to 4 times the amount secreted as lactose, which indicated values less than the calculated mammary glucose needs and that little lactose was synthesized. The excess glucose taken-up was used for other metabolic processes. Linear relationships between metabolite ${\Delta}AVs$ and arterial blood plasma concentration [A] showed that in Bali cows triglycerides (TG), phenylalanine (Phe) and tyrosine (Tyr) have high coefficients of determination, i.e. 0.77, 0.81 and 0.69, respectively. For glucose, the relationship is quadratic with an $R^2$ value of 0.49. It was concluded that lactose synthesis was inadequate, which led to a speculation that milk yield could be improved by increased lactose synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.6
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• pp.774-780
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• 2011

Inadequate supply of glucose or glucogenic substrates to the body tissues can affect metabolism of absorbed acetogenic metabolites from the gut and therefore, influence feed intake in ruminants. This study investigated the effect of energy supplementation on rumen kinetics in the gut, and the acetate clearance rate in the body tissues of sheep fed low quality basal roughage. A basal diet consisting of urea-treated mixture of wheaten chaff and barley straw (3:1 DM) containing 22.2 g N/kg DM was used. Four Merino cross wethers weighing $45{\pm}4.38\;kg$ fitted with permanent rumen and abomasal cannulae were allocated to four treatments in a $4{\times}4$ LSD. The treatments were basal diet ($E_0$), or basal diet supplemented with sucrose (112.5 g/d) administered intraruminally ($E_R$), abomasally ($E_A$), or via both routes (50:50) ($E_{RA}$). There was no difference (p>0.05) in the rumen liquid kinetics parameters between the four dietary treatments. However, there was a trend of animals supplemented with sucrose wholly or partly through the abomasum having lower faecal DM and therefore poor pellet formation, and low pH. Although the glucogenic potential of the fermentation products absorbed from the rumen was increased (p<0.001) by intra-ruminal supplementation with sucrose ($E_R$ and $E_{RA}$), there was no significant difference (p<0.05) in acetate clearance rate between the four dietary treatments.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.598-606
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• 2001

A mathematical model is proposed that can depict the kinetics of simultaneous saccharification and fermentation (SSF) using steam-exploded wood(SEW) with a glucose- and cellobiose-fermenting yeast strain. Brettanomyces custersii. An expression to describe the reduction of the relative digestibility during the hydrolysis of the SEW is introduced in the hydrolysis model. The fermentation model also takes two new factors into account, that is, the effects of the inhibitory compounds present in the SEW hydrolysates on the microorganism and the fermenting ability of Brettanomyces custersii, which can use both glucose and cellobiose as carbon sources. The model equations were used to simulate the hydrolysis of the SEW, the fermentation of the SEW hydrolysates, and a batch SSF, and the results were compared with the experimental data. The model was found to be capable of representing ethanol production over a range of substrate concentrations. Accordingly, the limiting factors in ethanol production by SSF under the high concentration of the SEW were identified as the effect of inhibitory compounds present in the SEW, the enzyme deactivation, and a limitation in the digestibility based on the physical condition of the substrate.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.4
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• pp.523-529
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• 2016

An isotope dilution method using $[U-^{13}C]glucose$ and $[1-^{13}C]leucine$ (Leu) was conducted to evaluate the effects of rice straw supplemented with urea and molasses (RSUM-diet) on plasma glucose and Leu turnover rates in sheep. Nitrogen (N) balance, rumen fermentation characteristics and blood metabolite concentrations were also determined. Four sheep were fed either mixed hay (MH-diet), or a RSUM-diet with a crossover design for two 21 days period. Feed allowance was computed on the basis of metabolizable energy at maintenance level. The isotope dilution method was performed as the primed-continuous infusion on day 21 of each dietary period. Nitrogen intake was lower (p = 0.01) for the RSUM-diet and N digestibility did not differ (p = 0.57) between diets. Concentrations of rumen total volatile fatty acids tended to be higher (p = 0.09) for the RSUM-diet than the MH-diet. Acetate concentration in the rumen did not differ (p = 0.38) between diets, whereas propionate concentration was higher (p = 0.01) for the RSUM-diet compared to the MH-diet. Turnover rates as well as concentrations of plasma glucose and Leu did not differ between diets. It can be concluded that kinetics of plasma glucose and Leu metabolism were comparable between the RSUM-diet and the MH-diet, and rumen fermentation characteristics were improved in sheep fed the RSUM-diet compared to the MH-diet.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.536-542
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• 2007

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.

• Korean Journal of Food Science and Technology
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• v.35 no.3
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• pp.417-422
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• 2003

The onions are considered to be a favorable functional source of beverage because they contain much sugar and various nutrients, and they are juicy vegetable. Recently, consumers have a new trend to take functional foods with health benefits. To meet this need, this study was the basic research to establish a manufacturing process of functional onion beverage by glucose oxidase. Glucose oxidase catalyzes reaction of glucose oxidation and makes generation of gluconic acid. Kinetics of the reaction was also investigated, and maximum glucose consumption rate $(V_{max})$ of $26.1{\times}10^{-2}\;g/L{\cdot}min$ and $K_m$ of 5.84 g/L were obtained. Optimum conditions were obtained when the glucose oxidase catalyzed reaction was carried out at temperature of $25^{\circ}C$, agitation rate of 450 rpm and aeration rate of 4 vvm in a 2.5 L jar fermentor. Finally, the enzyme reactor was 10-times scaled up and a similar glucose oxidation performance was achieved in the scaled-up reactor.

• Nutrition Research and Practice
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• v.3 no.2
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• pp.128-133
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• 2009

Few studies have examined short tenn responses to the different contents of carbohydrate or fat in the meal, although long tenn effects of the high fat meal have been considered as compound risk factor for metabolic disorders. The aim of this study was to investigate the postprandial changes of plasma glucose, insulin and lipids upon intakes of high carbohydrate or high fat meal in young healthy women. Subjects were randomly assigned to either the high carbohydrate meal (HCM, 75% carbohydrate, n=13) or the high fat meal (HFM, 60% fat, n=12) groups. The meals were prepared as isocaloric typical Korean menu. Blood samples were obtained prior to and 30, 60, 90, 120, 180 and 240 minute after the meal. There were no significant differences on fasting blood parameters including glucose, insulin, lipids concentrations between the groups prior to the test. The HCM had higher blood glucose and insulin concentrations, reached the peak at 30 min and maintained for 240 min compared to the HFM (P<0.05). The HFM had higher plasma triglyceride (TG) and free fatty acid (FFA) concentrations, reached the peak at 120 min and maintained for 240 min compared to the HCM (P<0.05). It is concluded that macronutrients content in the meal may be an important determinant of postprandial substrate utilization in healthy women.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.577-586
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• 2019

The engineered Aspergillus oryzae has a high NADPH demand for xylose utilization and overproduction of target metabolites. Glucose-6-phosphate dehydrogenase (G6PDH, E.C. 1.1.1.49) is one of two key enzymes in the oxidative part of the pentose phosphate pathway, and is also the main enzyme involved in NADPH regeneration. The open reading frame and cDNA of the putative A. oryzae G6PDH (AoG6PDH) were obtained, followed by heterogeneous expression in Escherichia coli and purification as a his6-tagged protein. The purified protein was characterized to be in possession of G6PDH activity with a molecular mass of 118.0 kDa. The enzyme displayed maximal activity at pH 7.5 and the optimal temperature was $50^{\circ}C$. This enzyme also had a half-life of 33.3 min at $40^{\circ}C$. Kinetics assay showed that AoG6PDH was strictly dependent on $NADP^+$ ($K_m=6.3{\mu}M$, $k_{cat}=1000.0s^{-1}$, $k_{cat}/K_m=158.7s^{-1}{\cdot}{\mu}M^{-1}$) as cofactor. The $K_m$ and $k_{cat}/K_m$ values of glucose-6-phosphate were $109.7s^{-1}{\cdot}{\mu}M^{-1}$ and $9.1s^{-1}{\cdot}{\mu}M^{-1}$ respectively. Initial velocity and product inhibition analyses indicated the catalytic reaction followed a two-substrate, steady-state, ordered BiBi mechanism, where $NADP^+$ was the first substrate bound to the enzyme and NADPH was the second product released from the catalytic complex. The established kinetic model could be applied in further regulation of the pentose phosphate pathway and NADPH regeneration of A. oryzae to improve its xylose utilization and yields of valued metabolites.

• KSBB Journal
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• v.16 no.1
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• pp.30-35
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• 2001

High cell density culture of Bacillus thuringiensis was conducted in fed-batch culture and TCRC using a bioreactor incorporating ceramic membrane filter. Cell growth of B. thuringiensis in fed-batch culture increased linearly, which was well matched by the results of cell growth modeling. In spite of the slower growth rate during fed-batch culture, no spore formation was observed, which was contrary to the results of continuous culture. Changing culture mode to batch culture after fed-batch operation induced a 2.7$\times$$10^9$ CFU/mL spore concentration using a 300 g/L glucose feed concentration. In TCRC operation incorporating ceramic filter within the bioreactor, the effect of glucose feed concentrations on the cell growth and spore formation of B. thuringiensis was determined. A maximum cell concentration of 1.8$\times$$10^{10}$ CFU/ml, which corresponds to 82.6 g-cell/L, was obtained in the TCRC using a 50 g/L glucose feed concentration. In the TCRC, cell growth increased linearly and glucose concentration was limited, which agreed well with the results of cell growth modeling. No spore formation was observed except when 1 g/L of glucose was fed. Changing to batch culture induced a 1.2$\times$$10^{10}$ CFU/mL of spore concentration, which was the highest spore concentration obtained among the various culture modes examined. The optimal glucose feed rate was found to be 0.55 g-glucose/h.

• Journal of Applied Biological Chemistry
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• v.42 no.1
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• pp.12-18
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• 1999

Sucrose synthase (EC 2.4.1.13) has been purified from the plant cytosolic fraction of chickpea (Cicer arietinum L. cv. Amethyst) nodules. The native enzyme had a molecular mass of $356{\pm}15kD$. The subunit molecular mass was $87{\pm}2kD$, and a tetrameric structure is proposed for sucrose synthase of chickpea nodule. Optimum activities in the sucrose cleavage and synthesis directions were at pH 6.5 and 9.0, respectively. The purified enzyme displayed typical hyperbolic kinetics with substrates in cleavage and synthesis reactions. Chickpea nodules sucrose synthase had a high affinity for UDP ($K_m$, $8.0{\mu}M$) and relatively low affinities for ADP ($K_m$, 0.23 mM), CDP ($K_m$, 0.87 mM), and GDP ($K_m$, 1.51 mM). The $K_m$ for sucrose was 29.4 mM. In the synthesis reaction, UDP-glucose ($K_m$, $24.1{\mu}M$) was a more effective glucosyl donor than ADP-glucose ($K_m$, 2.7 mM), and the $K_m$ for fructose was 5.4 mM. Divalent cations, such as $Ca^{2+}$, $Mg^{2+}$, and $Mn^{2+}$, stimulated the enzyme activity in both the cleavage and synthesis directions, and the enzyme was very sensitive to inhibition by $HgCl_2$ and $CuSO_4$.