Tyrosinases catalyze the hydroxylation of a monophenol (monophenolase activity) and the conversion of an o-diphenol to o-quinone (diphenolase activity), which are mainly involved in the modification of tyrosine residues into 3,4-dihydroxyphenyl-alanine (DOPA) and DOPA/DOPAquinone-derived intermolecular cross-linking. Previously, we obtained a slightly acidic and cold-active tyrosinase, tyrosinase-CNK, by our recombinant protein approach. The enzyme showed optimal activity at pH 6.0 and 20 ℃ with an abnormally high monophenolase/diphenolase activity ratio and still had approximately 50% activity compared with the highest activity even in ice water. Here, we investigated reaction stability of the recombinant tyrosinase-CNK as a psychrophilic enzyme. The enzyme showed remarkable thermal stability at 0 ℃ and the activity was well conserved in repeated freeze-thaw cycles. Although water-miscible organic solvent as reaction media caused the activity decrease of tyrosinase-CNK as expected, the enzyme activity was not additionally decreased with increased concentration in organic solvents such as ethanol and acetonitrile. Also, the enzyme showed high salt tolerance in chaotropic salts. It was remarkably considered that 2+ metal ions might inhibit the incorporation of Cu2+ into the active site. We expect that these results could be used to design tyrosinase-mediated enzymatic reaction at low temperature for the production of catechols through minimizing unwanted self-oxidation and enzyme inactivation.
Objective: A study was conducted to test six threonine (Thr) levels (0.39%, 0.44%, 0.49%, 0.54%, 0.59%, and 0.64%) to estimate the optimal dietary Thr requirements for Longyan laying ducks from 17 to 45 wk of age. Methods: Nine hundred Longyan ducks aged 17 wk were assigned randomly to the six dietary treatments, where each treatment comprised six replicate pens with 25 ducks per pen. Results: Increasing the Thr level enhanced egg production, egg weight, egg mass, and the feed conversion ratio (FCR) (linearly or quadratically; p<0.05). The Haugh unit score, yolk color, albumen height, and the weight, percentage, thickness, and breaking strength of the eggshell did not response to increases in the Thr levels, but the albumen weight and its proportion increased significantly (p<0.05), whereas the yolk weight and its proportion decreased significantly as the Thr levels increased. Conclusion: According to a regression model, the optimal Thr requirement for egg production, egg mass, and FCR in Longyan ducks is 0.57%, while 0.58% is the optimal level for egg weight from 17 to 45 wk of age.
This experiment was carried out to investigate the growth characteristics of virus-free sweetpotato in hydroponic bag culture. The rooted one-node and upper-shoot cuttings of 'Yeonhwangmi' and 'Mannamiwere' transplanted into the plastic bag (0.5 mm of thickness, $W300{\times}L1,200{\times}H120mm$) culture system filled with commercial horticultural media (TKS-2, Flora Gard Co.) and cocopeat + perlite (3 : 7, 5 : 5, 7 : 3, (v/v). Nutrient solution of National Horticultural Research Station in Japan was separately irrigated $3times{\times}30minutes$ per day (10:00, 14:00 and 20:00 with timer control) by the drip irrigation. The growth of two varieties was not significantly different among four substrates in 15 days, but it was excellent in commercial horticultural media (TKS-2) and cocopeat 3 : perlite 7 (v/v) in 20 days. The growth in 'Yeonhwangmiwas' promoted than that in 'Mannami', and the upper-shoot cuttings of 'Yeonhwangmi' and 'Mannami' showed a significant vine elongation (over 10 and 2 cm, respectively) compared to the one-node cuttings. Fresh weight and dry weight in cocopeat 3 : perlite 7 (v/v) were significant increased than that in cocopeat 7 : perlite 3 (v/v). Therefore, the bag culture system filed with cocopeat 3 : perlite 7 (v/v) was successfully applied on propagation of virus-free sweetpotato seedling, and the result expected that the bag culture system could be commercialized without high financial costs for farmers.
Kim, Jung-Hyun;Lee, Chang-Bo;Baek, Seung-Wook;Park, Kwang-Jin;Bae, Joong-Myeon
한국신재생에너지학회:학술대회논문집
/
2007.06a
/
pp.196-199
/
2007
The chemical states of oxygen on the surfaces of $Pr_{1-x}Sr_{x}CoO_{3}$ (x=0.5 and 0.7) oxide systems were investigated by X-ray photoelectron spectroscopy. Merged oxygen peaks of $Pr_{1-x}Sr_{x}CoO_{3}$ (x=0.5 and 0.7) oxides could be divided as five sub-peaks. These five sub-peaks could be defined as lattice oxygen ($O_{L}$). chemisorbed oxygen peaks ($O_{C}$) and hydroxyl condition oxygen peak ($O_{H}$). In case of the $Pr_{0.5}Sr_{0.5}CoO_{3}$ and $Pr_{0.3}Sr_{0.7}CoO_{3}$, the binding energy (BE) of oxygen lattice were located at same BE. However, the BE of chemisorbed oxygen peaks including oxygen vacancy shows different BE. Especially, it was found that BE of chemisorbed oxygen peaks was increased when more Sr were substituted. Comparing atomic percentages of oxygens of $Pr_{0.5}Sr_{0.5}CoO_{3}$ and $Pr_{0.3}Sr_{0.7}CoO_{3}$, the ratio of $Pr_{0.3}Sr_{0.7}CoO_{3}$ was higher than that of $Pr_{0.5}Sr_{0.5}CoO_{3}$. It showed more chemically adsorbed site including oxygen vacancies were existed in $Pr_{0.3}Sr_{0.7}CoO_{3}$.
An experiment was conducted to study the zinc deficiency of rice plant grown on Boeun soil of Korea and on Batangas soil of the philippines. The results obtained are summarized as follows: 1. Batangas soil was deficient in zinc because of the high pH, high organic matter and high calcium. Zinc content of the rice plant grown on Batangas soil was l6ppm. On the other hand, Boeun soil had low organic matter. Zinc content of the rice plant grown on Boeun soil was 33ppm. 2. The top leaf had higher zinc content than any other leaves. Zinc deficiency apparently occured when the zinc content of the rice plant was below 18ppm. 3. The physiologically acid fertilizer such as ammonium sulfate was more advantageous in improving zinc up take than the urea. 4. The zine deficiency was promoted when the Mn/Zn ratio is high, and zinc and manganess interact antagonistically in the growth of rice plant. 5. The zinc deficiency could be overcome by applying zinc chloride and fritted Trace elements to the Batangas soil, and the yield was significantly increased than conrtol.
The strain A49, which produces a new type of extracellular polysaccharide was isolated from soil samples. From morphological, physiological and biochemical tests, the strain A49 was identified as a Bacillus polymyxa and named Bacillus polymyxa A49. Bacillus polymyxa A49 was found to produce a highly viscous extracellular polysaccharide when grown aerobically in a medium containing glucose as the sole source of carbon. The polysaccharide (A49 POL) showed a homogeneous pattern on gel permeation chromatography (GPC) and its molecular weight was estimated to be about 1.6 mega dalton (mDa). The FT-IR spectrum of A49-POL revealed typical characteristics of polysaccharides. As a result of investigations with HPLC and carbozole assay, A49-POL was found to consist of L-fucose, D-galactose, D-glucose, D-mannose, and D-glucuronic acid, with the molar ratio of these sugars being approximately 1:2:7:50:12. Rheological analysis of A49 POL revealed that it is pseudoplastic and has a higher apparent viscosity at dilute concentrations than does xanthan gum. The consistancy factor of A49 POL was found to be higher, and the flow index of A49 POL lower, than xanthan gum. Its apparent viscosity was comparatively unstable at various temperatures. the A49 POL showed the highest apparent viscosity at pH 3. When salts were added to A49 POL solution, the solution was compatible with up to 10% KCl, 35% NaCl, 55% $CaCl_2$, 55% $MgCl_2$, 55% $K_2HPO_4$, and 110% $Ca({NO_3})_2$, respectively.
Kim, Kyu-Sik;Kim, Bok-Young;Lee, Min-Hyo;Han, Ki-Hak;Kim, Maun-Soo
Korean Journal of Environmental Agriculture
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v.4
no.2
/
pp.102-107
/
1985
A pot experiment was conducted to find out the effects of water management, slaked lime and wollastonite on growth and Cu uptake of rice at Cu added soil. The soil was adjusted to 0, 50, 100 and 200 ppm concentration of Cu. The application amount of slaked lime was the lime requirement plus 150 ㎏/l0a and wollastonite 200 ㎏/10a, respectively. The copper concentration in soil which reduced yield significantly was 133.1 ppm for submersion and 136.8 ppm for intermittent irrigation. The application of lime and wollastonite reduced Cu content in brown rice as well as increased rice yields compared to that of no lime. The copper content in plant was increased with increasing soil Cu concentration, however, reduced with submersion and application of slaked lime, and increased with increasing the ratio of Cu/Ca+Mg equivalent in soil.
The production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) are known to be modulated by a variety of factors. Recent study showed that endotoxin-induced NO synthesis and iNOS expression were greatly enhanced by elevation of extracellular glucose concentration in murine macrophages. Although this was suggested to be due to the activation of protein kinase C (PKC) via sorbitol pathway, there was lack of evidence for this speculation. This study was performed to delineate the underlying intracellular mechanisms of glucose-enhancing effect on endotoxin-induced NO production in Raw264.7 macrophages. The levels of NO release induced by lipopolysaccharide (LPS) significantly increased by the treatment of glucose in a concentration dependent manner and also, this effect was observed in LPS-preprimed cells. Concurrent incubation of cells with PKC inhibitors, H-7 or chelerythrine, and LPS resulted in the diminution of NO production regardless of glucose concentration but this was not in the case of LPS-prepriming, that is, chelerythrine showed a minimal effect on the glucose- enhancing effect. PMA, a PKC activator, did not show any significant effect on glucose-associated NO production. Modulation of sorbitol pathway with zopolrestat, an aldose reductase inhibitor, did not affect LPS-induced NO production and iNOS expression under high glucose condition. And also, sodium pyruvate, which is expected to normalize cytosolic $NADH/NAD^+$ ratio, did not show any significant effect at concentrations of up to 10 mM. Glucosamine marginally increased the endotoxin-induced nitrite release in both control and high glucose treated group. 6-diazo-5-oxonorleucine (L-DON) and azaserine, glutamine: fructose- 6-phosphate amidotransferase (GFAT) inhibitors, significantly diminished the augmentation effect of high glucose on endotoxin-induced NO production. On the other hand, negative modulation of GFAT inhibitors was not reversed by the treatment of glucosamine, suggesting the minimal involvement, if any, of glucosamine pathway in glucose-enhancing effect. In summary, these results strongly suggest that the hexosamine biosynthesis pathway and the activation of PKC via sorbitol pathway do not contribute to the augmenting effect of high glucose on endotoxin induced NO production in macrophage-like Raw264.7 cells.
Hayirli, Armagan;Esenbuga, N.;Macit, M.;Lacin, E.;Karaoglu, M.;Karaca, H.;Yildiz, L.
Asian-Australasian Journal of Animal Sciences
/
v.18
no.9
/
pp.1310-1319
/
2005
This experiment was conducted to determine the effects of cage density (CD) and humate supplementation (HS) on laying performance, metabolic profile, and egg quality during the peak production period in hens. Lohman layers (n = 180, 46 wks of age) were blocked according to the location of cages and then allocated randomly to two levels of CD (4 or 6 hens per cage or 540 vs. 360 $cm^2$/hen) and three levels of HS (0, 0.15, and 0.30%). Egg production (EP) and feed consumption (FC) were measured daily; egg weight was measured bi-weekly; and BW was measured before and after the experiment. Blood and additional egg samples were obtained at the end of the experiment for determination of metabolic profile and egg quality. The data were analyzed using two-way ANOVA as repeated measures. Except for FC, CD did not affect laying performance parameters. Hens placed in high-density cages had lower FC than hens placed in normal-density cages. Increasing HS level linearly increased FC, EP, and feed conversion ratio (FCR). There was a CD by HS interaction effect on FC and EP. Hens placed in high-density cages had greater serum glucose, total protein, albumin, globulin, Ca, and P concentrations and tended to have greater serum corticosterone concentration than hens placed in normaldensity cages. Increasing HS level linearly increased serum glucose, total protein, albumin, globulin, creatine, and Ca concentrations and linearly decreased serum triglyceride and very low-density lipoprotein concentrations. There was a CD by HS interaction effect on serum glucose and albumin concentrations. There were no alterations in egg quality parameters in response to increasing CD. Albumen index and Haugh unit decreased linearly and other egg quality parameters did not change as HS level increased. In conclusion, increased caging density adversely affected metabolic profile, despite insignificantly deteriorating laying performance. Moreover, benefits from humate supplementation seem to be more noteworthy for hens housed in stressing conditions than for hens housed in standard conditions.
Chung, Suk-Jae;Ramanathan, Vikram;Brett, Claire M.;Giacomini, Kathleen M.
Journal of Pharmaceutical Investigation
/
v.25
no.3
/
pp.7-20
/
1995
Taurine, a ${\beta}-amino$ acid, plays an important role as a neuromodulator and is necessary for the normal development of the brain. Since de novo synthesis of taurine in the brain is minimal and in vivo studies suggest that taurine dose not cross the blood-brain barrier, we examined whether the choroid plexus, the blood-cerebrospinal fluid (CSF) barrier, plays a role in taurine transport in the central nervous system. The uptake of $[^3H]-taurine$ into ATP depleted choroid plexus from rabbit was substantially greater in the presence of an inwardly directed $Na^+$ gradient taurine accumulation was negligible. A transient in side-negative potential gradient enhanced the $Na^+-driven$ uptake of taurine into the tissue slices, suggesting that the transport process is electrogenic, $Na^+-driven$ taurine uptake was saturable with an estimated $V_{max}$ of $111\;{\pm}\;20.2\;nmole/g/15\;min$ and a $K_M\;of\;99.8{\pm}29.9\;{\mu}M$. The estimated coupling ratio of $Na^+$ and taurine was $1.80\;{\pm}\;0.122.$$Na^+-dependent$ taurine uptake was significantly inhibited by ${\beta}-amino$ acids, but not by ${\alpha}-amino$ acids, indicating that the transporter is selective for ${\beta}-amino$ acids. Since it is known that the physiological concentration of taurine in the CSF is lower than that in the plasma, the active transport system we characterized may face the brush border (i.e., CSF facing) side of the choroid plexus and actively transport taurine out of the CSF. Therefore, we examined in vivo elimination of taurine from the CSF in the rat to determine whether elimination kinetics of taurine from the CSF is consistent with the in vitro study. Using a stereotaxic device, cannulaes were placed into the lateral ventricle and the cisterna magna of the rat. Radio-labelled taurine and inulin (a marker of CSF flow) were injected into the lateral ventricle, and the concentrations of the labelled compounds in the CSF were monitored for upto 3 hrs in the cisterna magna. The apparent clearance of taurine from CSF was greater than the estimated CSF flow (p<0.005) indicating that there is a clearance process in addition to the CSF flow. Taurine distribution into the choroid plexus was at least 10 fold higher than that found in other brain areas (e. g., cerebellum, olfactory bulb and cortex). When unlabelled taurine was co-administered with radio-labelled taurine, the apparent clearance of taurine was reduced (p<0.0l), suggesting a saturable disposition of taurine from CSF. Distribution of taurine into the choroid plexus, cerebellum, olfactory bulb and cortex was similarly diminished, indicating that the saturable uptake of taurine into these tissues is responsible for the non-linear disposition. A pharmacokinetic model involving first order elimination and saturable distribution described these data adequately. The Michaelis-Menten rate constant estimated from in vivo elimination study is similar to that obtained in the in vitro uptake experiment. Collectively, our results demonstrate that taurine is transported in the choroid plexus via a $Na^+-dependent,saturable$ and apparently ${\beta}-amino$ acid selective mechanism. This process may be functionally relevant to taurine homeostasis in the brain.
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