Mitsuhashi, T.;Mitsumoto, M.;Yamashita, Y.;Ozawa, S.
Asian-Australasian Journal of Animal Sciences
/
v.1
no.2
/
pp.99-106
/
1988
A high performance liquid chromatographic procedure is described for the direct determination of the picomole amount of palmitoyl-Coenzyme A and stearoyl-Coenzyme A, using a stainless steel column packed with C-18 derivatized porous silica ($5{\mu}m$), an isocratic elution with a mixture of 33 mM $KH_2PO_4$/acetonitrile as a mobile phase and a UV detector. The long-chain acyl-Coenzyme A esters were determined in incubated microsomal fractions of a bovine liver to demonstrate the utility of this method for monitoring acyl-CoA synthesis in biological samples. The reaction rate of palmitate was higher than that of stearate. After a 60 minute incubation period, the generated amount of palmitoyl-Coenzyme A and stearoyl-Coenzyme A were approximately 70 and 20 n mol/mg micresomal protein, respectively. The advantage of this method are in that no decomposition of the CoA esters is involved, while the constituent molecular species is detected.
The soil of Kwangneung area(Kyeunggi-Do) was inoculated directly into wheat-bran-media and after $3{\sim}4$ days of incubation, a Penicillium species whose cellulase activity was 1011u/g was isolated. With the treatment of mutagenic agents an improved strain(cellulase activity: 1303u/g) was obtained. This strain was screened again by mono-spore isolation method. Finally a strain C8-14 (cellulase activity: 2351u/g) which had lesser spores than the wild strain was obtained.
Objective: This study explored the physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed (IOF) supplement. Methods: Amniotic fluid was collected from broiler breeders (Ross 308, 51 weeks and Cobb 500, 35 weeks) on day 17 after incubation. A mixture of high-quality soy protein supplement - Hamlet Protein AviStart (HPA) was serially diluted in MilliQ water to obtain solutions ranging from 150 to 9.375 mg/mL. The mixtures were heat-treated (0, 30, 60 minutes) in a waterbath ($80^{\circ}C$) and then centrifuged to obtain supernatants. The amniotic fluid and HPA supernatants were analysed for their physico-chemical properties. Results: Only viscosity and $K^+$ were significantly (p<0.05) different in both strains. Of all essential amino acids, leucine and lysine were in the highest concentration in both strains. The osmolality, viscosity and $pCO_2$ of the supernatants decreased (p<0.05) with decreasing HPA concentration. Heat treatment significantly (p<0.05) affected osmolality, pH, and $pCO_2$, of the supernatants. The interactions between HPA concentration and heat treatment were significant with regards to osmolality (p<0.01), pH (p<0.01), $pCO_2$ (p<0.05), glucose (p<0.05), lactate (p<0.01) and acid-base status (p<0.01) of HPA solutions. The $Ca^{2+}$, $K^+$, glucose, and lactate increased with increasing concentration of HPA solution. The protein content of HPA solutions decreased (p<0.05) with reduced HPA solution concentrations. The supernatant from 150 mg/mL HPA solution was richest in glutamic acid, aspartic acid, arginine and lysine. Amino acids concentrations were reduced (p<0.05) with each serial dilution but increased with longer heating. Conclusion: The values obtained in the primary solution (highest concentration) are close to the profiles of high-protein ingredients. This supplement, as a solution, hence, may be suitable for use as an IOF supplement and should be tested for this potential.
Kim, Tae Jin;Hwang, Hyun Young;Hong, Chang Oh;Lee, Jeung Joo;Kim, Gun Yeob;Kim, Pil Joo
Korean Journal of Environmental Agriculture
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v.33
no.4
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pp.282-289
/
2014
BACKGROUND: Methane($CH_4$) is considered as the secondmost potent greenhouse gas after carbon dioxide ($CO_2$). Methanogenesis is an enzyme-mediated multi-step process by methanogens. In the penultimate step, methylated Co-M is reduced by methyl Co-M reductase (MCR) to $CH_4$ involving a nickel-containing cofactor F430. The activity of MCR enzyme is dependent on the F430 and therefore, the bioavailability of Ni to methanogens is expected to influence MCR activity and $CH_4$ production in soil. In this study, different doses of EDTA(Ethylene Diamine Tetraacetic Acid) were applied in flooded soils to evaluate their suppression effect on methane production by chelating Ni of methanogenesis cofactor. METHODS AND RESULTS: EDTA was selected as chelating agents and added into wetland and rice paddy soil at the rates of 0, 25, 50, 75, and $100mmol\;kg^{-1}$ before 4-weeks incubation test. During the incubation, cumulative $CH_4$ production patterns were characterized. At the end of the experiment, soil samples were removed from their jars to analyze total soil Ni and water-soluble Ni content and methanogen abundance. Methane production from 100 mmol application decreased by 55 and 78% in both soils compared to that from 0 mmol. With increasing application rate of EDTA in both soils, water-soluble Ni concentration significantly increased, but total soil Ni and methanogen activities showed negative relationship during incubation test. CONCLUSION: The decrease in methane production with EDTA application was caused by chelating Ni of coenzyme F430 and inhibiting methanogenesis by methyl coenzyme M reductase. Consequently, EDTA application decreased uptake of Ni into methanogen, subsequently inhibited methanogen activities and reduced methane production in flooded soils.
The effects of punching treatment on mycelial culture and fruiting body productivity were investigated in a new Lentinula edodes cultivar, "Jadam", in sawdust medium for the stable production of oak mushroom. As the punching volume and number increased, the weight loss rate and color difference increased and the L value decreased. After spawn inoculation, the sawdust medium temperature and CO2 concentration reached their highest values at 33 and 19 days of incubation, respectively. The O2 concentration showed the lowest value on the 14th day of incubation, which was the opposite pattern to the CO2 concentration. As the punching volume and the number increased, the medium temperature and O2 concentration increased, and the CO2 concentration decreased. Higher punching volumes and numbers resulted in higher temperatures and lower CO2 concentrations. The best fruiting body yield was 5 × 70 mm - 30 (punching diameter × depth - number), and the total yield after three cycles was 644.7 g.
Journal of The Korean Society of Grassland and Forage Science
/
v.32
no.1
/
pp.59-74
/
2012
Buffer solubility and protein fractionation were evaluated from the hays (timothy, alfalfa and klein) and straws (tall fescue and rice), and $In$$vitro$ trial was conducted to examine the effect of buffer extraction on fermentation characteristics, degradability and gas ($CO_2$ and $CH_4$) production. Buffer soluble protein (SP) content and A fraction in total protein were highest in alfalfa hay as 61% and 41.77%, respectively while lowest in rice straw (42.8% and 19.78%, respectively). No difference was observed in B1 fraction among forages but B2 fraction was slightly increased in klein hay (12.34%) and tall fescue straw (10.05%) compared with other forages (6.34~8.85%). B3 fraction of tall fescue was highest as 38.49% without difference among other forages while C fraction was highest in rice straw. pH in incubation solution was higher in all forages after extraction than before extraction at 3h (P<0.01) and 6h (P<0.05), and pH from hays of timothy and alfalfa was higher than the other forages at 6h (P<0.05) and 12h (P<0.001). Regardless of extraction, ammonia-N concentration from alfalfa hay was increased at all incubation times and extraction effect was appeared only at 3h incubation time (P<0.01). Total VFA concentration from alfalfa hay was highest up to 24h incubation while those from tall fescue straw and rice straw were lowest. Buffer extraction decreased (P<0.01~P<0.001) the total VFA concentration. Acetic acid proportion was increased (P<0.001) before extraction of forages but no difference was found between forages. Propionic acid($C_3$) proportion was also increased(P<0.001) before extraction in all forages than in straws at 3h, 24h and 48h incubations, and $C_3$ from hays were mostly higher (P<0.05) than from straws. Butyric acid proportion, however, was not affected by extraction at most incubation times. Parameter 'a' regarding to the dry matter (DM) degradation was increase (P<0.001) in all forages before extraction, and was decreased (P<0.05) in tall fescue straw and rice straw compared with hays. Parameter 'b' was also increased (P<0.001) before extraction but no difference was found between forages. Effective degradability of DM (EDDM) was higher (P<0.001) before extraction in most forages except for rice straw. Buffer extraction decreased (P<0.05) all parameters (a, b, and c) regrading to the crude protein (CP) degradation but no difference was found between forages. Effective degradation of CP (EDCP) was lower (P<0.05) in straws than in hays. Parameters 'a' and 'b' regarding to the NDF degradation (P<0.01) and effective degradability of NDF (EDNDF, P<0.001) were also higher in forages before extraction than after extraction but no difference was found between forages. Buffer extraction reduced (P<0.05~P<0.001) $CO_2$ production from all the forages uo to 24h incubation and its production was greater (P<0.05~P<0.01) from hays than straws. Methane ($CH_4$) production was also greater (P<0.01~P<0.001) in all forages at all incubation times, and its production was greater (P<0.05) from hays than from straws at most incubation times. Based on the results of the current study, it can be concluded that buffer solubility and CP fractionation might be closely related with $In$$vitro$ VFA concentration, degradability and gas ($CO_2$ and $CH_4$) production. Thus, measurement of buffer solubility and protein fractionation of forages might be useful to improve TMR availability in the ruminants.
The metabolic patterns of C-1 and C-6-carbon atoms of glucose were observed in the tissue homogenates of the Ehrlich ascites tumor tissue which was incubated for 3 hours in the Dubnuff metabolic shaking incubator. $C^{14}-1-and\;C^{14}-6-glucose$ were used as tracers. The glucose media in which tissue homogenate was incubated was kept at a concentration of 200mg% glucose of carrier and appropriate amount of $C^{14}-1-or\;C^{14}-6-tracer$. At the end of 3 hour incubation, respiratory $CO_2$ samples trapped by alkaline which is placed in the tenter well of incubation flask were analyzed for the total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate, pyruvate and glycogen and calculations were made on the glucose consumption rate, pyruvate and lactate accumulation rates. The following results were obtained. Data obtained in each group are as follows: 1. In the tissue homogenate, which was incubated with $C^{14}-1-glucose as a substrate, total $CO_2$ production rate averaged $19.0{\pm}5.0{\mu}M/hr/gm$ and the mean specific activity of respiratory $CO_2$ was $840{\pm}296\;cpm/mgC.$ Relative specific activity (RSA) which means the fraction of $CO_2$ derived from medium $C^{14}-1-glucose$ to total $CO_2$ production rate was calculated by ratio of SA of respiratory $CO_2$ and medium $C^{14}-1-glucose.$ RSA was $14.3{\pm}5.0%,$ Accordingly actual $CO_2$ production rate from medium $C^{14}-1-glucose$ showed a mean value of $2.79{\pm}1.35\;{\mu}m$ of which amount was equivalent to the mean value of total glucose consumption rate $(RGDco_2)$, namely, $5.1{\pm}1.3%.$ Lactate and pyruvate appearance rates averaged $7.13{\pm}1.26\;and\;0.21{\pm}0.02{\mu}M/hr/gm,$ respectively. Assuming that these 3 carbon compounds appeared in the medium were derived from glucose, calculations were made that relative glucose disappearance rate into lactate $(RGD_L)$ was $38.0{\pm}5.4%\;and\;RGD_P$ was $1.23{\pm}0.03%.$ Therefore, about 43.3% of the total glucose consumed were accounted for by conversion into the respiratory $CO_2$, lactate and pyruvate. 2. In the second group, which was incubated with $C^{14}-1-glucose$ as a substrate, glucose consumption rate, lactate and pyruvate appearance rates showed almost the same order as the values of the $C^{14}-1-glucose$ substrate group. However, RSA was remarkably decreased showing a mean value of $1.02{\pm}0.13%.$ This fact means that the C-6 carbon of glucose take the minor part in the oxidative metabolism of glucose. The glycogen level in both substrate tissue homogenate showed less than 0.3% of tissue weight. These low value suggested that there was an inhibition of carbohydrate synthesis in the Ehrlich ascites tumor tissue. 3. The catabolic pathway of glucose in the tumor tissue were analyzed on the basis of Bloom's principle from the values of RSA. It was found that in the tumor tissue more than 90% of $CO_2$ derived from glucose were oxidized via the alternate pathway other than principal EMP-TCA cycle such as hexose monophosphate pathway (HMP). From the data described above, it was assumed that in the Ehrlich tumor tissue anaerobic glycolysis proceeds normally although, the oxidation of products of anaerobic glycolysis via the TCA cycle is inhibited resulting in the accumulation of lactate and almost all of oxidative energy from glucose is released by oxidative pathway such as HMP.
Kim, You Jin;Park, Han;Kim, Min-Ho;Seo, Sung Hee;Ok, Yong Sik;Yoo, Gayoung
Journal of Korean Society of Environmental Engineers
/
v.37
no.7
/
pp.432-440
/
2015
Biochar, a by-product from pyrolysis of biomass, is a promising option to mitigate climate change by increasing soil carbon sequestration. This material is also considered to have potential to remediate a soil with heavy metal pollution by increasing the soil's adsorptive capacity. This study conducted the assessment of two biochars considering the climate change mitigation potential and heavy metal removal capacity at the same time. Two kinds of biochars (BC_Ch, TW_Ch) were prepared by pyrolyzing the biomass of burcucumber (BC_Bm) and tea waste (TW_Bm). The soils polluted with Pb were mixed with biochars or biomass and incubated for 60 d. During the incubation, $CO_2$, $CH_4$, and $N_2O$ were regularly measured and the soil before and after incubation was analyzed for chemical and biological parameters including the acetate extractable Pb. The results showed that only the BC_Ch treatment significantly reduced the amount of Pb after 60 d incubation. During the incubation, the $CO_2$ and $N_2O$ emissions from the BC_Ch and TW_Ch were decreased by 24% and 34% compared to the BC_Bm and TW_Bm, respectively. The $CH_4$ emissions were not significantly affected by biochar treatments. We calculated the GWP considering the production of amendment materials, application to the soils, removal of Pb, and soil carbon storage. The BC_Ch treatment had the most negative value because it had the higher Pb adsorption and soil carbon sequestration. Our results imply that if we apply biochar made from burcucumber, we could expect the pollution reduction and climate change mitigation at the same time.
To investigate the contribution of macroalgae to biogeochemical nutrients and carbon cycles, we measured the uptake rates of nutrients and $CO_2$ by Undaria pinnatifida using an incubation method in an acrylic chamber. From January to March 2010, U. pinnatifida was sampled at Ilkwang, a well-known area of macroalgae culture in Korea. The initial and final concentrations of nutrients, dissolved oxygen, total alkalinity, and pH of the chamber water were measured, and production/uptake rates were calculated using concentration changes, chamber volume, and incubation time. The production rate of dissolved oxygen by U. pinnatifida (n = 32) was about $5.4{\pm}4.0\;{\mu}mol\;g_{fw}^{-1\;}h^{-1}$. The uptake rate of total dissolved inorganic carbon (TDIC), calculated by total alkalinity and pH, was $7.9{\pm}6.5\;{\mu}mol\;g_{fw}^{-1}\;h^{-1}$. Nutrients uptake averaged $141.7{\pm}119.2$ nmol N $g_{fw}^{-1}\;h^{-1}$ and $15.0{\pm}9.1$ nmol P $g_{fw}^{-1}\;h^{-1}$. A positive linear correlation ($r^2$ = 9.6) existed between the production rate of dissolved oxygen and the uptake rate of total dissolved inorganic carbon, suggesting that these two factors serve as good indicators of U. pinnatifida photosynthesis. The relationships between fresh weight and uptake rates of nutrients and $CO_2$ suggested that younger specimens (<~50 g fresh weight) are much more efficient at nutrients and $CO_2$ uptake than are specimens >50 g. The amount of carbon uptake by the total biomass of U. pinnatifida in Korea during the year of 2008 was about 0.001-0.002% of global ocean carbon uptake. Thus, more research should be focused on macroalgae-based biogeochemical cycles to evaluate the roles and contributions of macroalgae to the global carbon cycle.
In order to clarify how much of the residues of Bentazon could be taken up by crops, soybean and radish were grown for 28 days in soils containing freshly treated $^{14}C-Bentazon$ and non-extractable soil-hound residues of $^{14}C-Bentazon.$ The results obtained are summarized as follows. 1. $^{14}CO_2$ evolution from $^{14}C$-Bentazon during the 6-month pre-incubation in soil was 14.79% relative to the applied radioactivity. 2. Mineralization of ^$^{14}C$-Bentazon in soil to $^{14}CO_2$ during 28 days of crop growing was much higher in the freshly treated soil than in the bound soil, and much higher in radish than in soybean. 3. The amounts of $^{14}C-Bentazon$ and its metabolites absorbed by soybean and radish were 45.41 and 21.48%, respectively, in freshly treated soil, whereas those were 3.92 and 1.23% in bound soil, respectively. The translocation ratios of radioactivity .from the root to the shoot were much higher in radish than in soybean, remarkably. 4. The uptake ratios of the freshly treated $^{14}C-Bentazon$ to the bound $^{14}C-Bentazon$ by soybean and radish were 12 : 1 and 17 : 1, respectively. 5. It was well verified that the presence of crops enhanced the mineralization to $^{14}CO_2$ and the transformation to polar metabolites of Bentazon.
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