• Asian-Australasian Journal of Animal Sciences
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• 제23권9호
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• pp.1198-1204
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• 2010

This study was conducted to investigate the effects of dietary supplementation with multiple enzymes composed of phytase plus carbohydrolases (ROVABIO$^{(R)}$ Max, RM) on growth performance, nutritional availability and intestinal viscosity in broiler chicks. A total of one thousand, one-day-old male broiler chicks were randomly allotted into treatment groups that received one of five experimental diets for 32 days. Each group consisted of 40 birds and all experiments included five replicates. The dietary treatments included PC (a positive control diet), NC1 (65 kcal/kg, 0.15% and 0.10% less ME, available phosphorus and calcium levels, respectively, than the PC diet), NC2 (85 kcal/kg, 0.20% and 0.10% less ME, available phosphorus and calcium levels, respectively, than the PC diet), NC1+RM (NC1 plus ROVABIO$^{(R)}$ Max) and NC2+RM (NC2 plus ROVABIO$^{(R)}$ Max). The average body weights, daily body weight gains and feed conversion rates of the chicks fed a diet containing RM improved significantly or tended to improve. The treatments also had no effect on the carcass characteristics or blood parameters, but the viscosity of the intestinal contents of the chicks fed the diet containing RM was significantly lower than that of chicks in the NC without RM groups. Additionally, chicks fed the dietary RM showed increased breaking strength and ash content of the tibia when compared to chicks that received the non-RM diets. Taken together, the results of the present study indicated that the addition of multiple enzymes consisting of phytase plus NSP enzymes improved the growth performance and mineral status of the tibia in broiler chickens fed corn-wheat-soybean meal-based diets with reduced levels of nutrients. Further, these findings suggest that the improved animal performance is associated with reduced intestinal viscosity by the dietary enzyme complex.

• 대한유전성대사질환학회지
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• 제15권2호
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• pp.72-77
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• 2015

Purpose: Deficits of the respiratory chain are reported to be the major cause of Leigh syndrome is said to be the underlying causes. The need for biochemical diagnosis to draw more accurate diagnosis or prognosis to support treatments is rapidly increasing. This study tried to analyze the aspects of clinical characteristics and biochemical diagnosis of mitochondrial respiratory chain complex (MRC) defect in Leigh syndrome, using methods of biochemical enzyme assay. Methods: We included total number of 47 patients who satisfied the clinical criteria of Leigh syndrome and confirmed by biochemical diagnosis. All those patients went through muscle biopsy to perform biochemical enzyme assay to analyze MRC enzyme in order to find the underlying cause of Leigh syndrome. Results: MRC I defect was seen in 23 (48.9%) cases taking the first place and MRC IV defect in 15 (31.9%) following it. There were 9 (19.2%) cases of combined MRC defect. Combined cases of type I and IV were detected in 7 (14.9%) patients while type I and V in 2 (4.3%). The onset age of symptom was less than 1 year old in 28 (59.6%). The most common early symptom, observed in 23 (48.9%), was delayed development, but there were other various neurological symptoms observed as well. In regard with the disease progression, 35 (74.5%) patients showed slowly progressive course, the one that progressed continuously but slowly over 2 years of period. As for Maximum motor development, 22 (46.8%) were bed-ridden state, most of them suffering serious delayed development. Patients showed various symptoms with different organs involved, though neuromuscular involvement was most prominent. Delayed development was seen in all cases. Multifocal lesion in brain MRI study was seen in 36 (76.6 %) cases, taking a greater percentage than 11 (23.4%) cases with single lesion. In MR spectroscopy study, the characteristic lactate peak of mitochondrial disease was identified in 20 (42.6%) patients. Conclusions: Further analysis of clinical and biochemical diagnosis on more extended group of patients with Leigh syndrome will enable us to improve diagnostic precision and to understand the natural course of mitochondrial disease.

• Journal of the Korean Wood Science and Technology
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• 제38권3호
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• pp.251-261
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• 2010

Formiptosis pinicola KMJ812에 의해 생산된 조효소액은 섬유소 분해효소 복합체로서 매우 활성이 높으며 특히 ${\beta}$-glucosidase의 활성이 높아 포도당 전환수율이 높다. 본 연구에서는 F. pinicola KMJ812 생산 cellulase를 고정화에 따른 효소특성의 변화와 고정화 효소의 재사용에 따른 효소의 불활성 정도를 관찰하였다. 담체는 고정화 수율이 cellulase활성 61.7%와 ${\beta}$-glucosidase활성 64.4%로 우수한 Duolite A568로 선정하였다. 고정화 효소의 최적반응온도는 cellulase와 ${\beta}$-glucosidase 활성의 경우 모두$55^{\circ}C$로서 수용성효소보다 높았으며, 최적 pH는 cellulase활성은 4.0이었고, ${\beta}$-glucosidase활성은 4.5로 cellulase활성의 경우에서만 수용성효소와 비교하여 약간 염기성으로 변화하였다. 본 고정화 효소는 $50^{\circ}C$에서 72시간 후에 98%의 활성을 유지하고 있었으며, $50^{\circ}C$에서 8회 사용 후에 50%정도의 잔존활성을 나타내었다.

• Applied Biological Chemistry
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• 제14권3호
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• pp.229-235
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• 1971

우리나라에서 재배되고 있는 대표적인 수도 품종 중에서 japonica type으로 관옥, 후지사까 5호, 팔달, 수원 82호의 4품종과 indica type으로 IR-262 CP-slo의 2품종을 선정하여 estel형 인 유기인계 살충제 malathion과 방향족 ester로서 p-nitrophenyl acetate에 대한 esterase의 활성을 비교해 보고 agar-gel electrophoresis에 의하여 ${\alpha}-naphthyl$ acetate를 기질로 하여 esterase의 zymogram을 관찰한 바를 요약해 보면 다음과 같다. 1. 조효소액(crude enzyme)의 malathion가수분해 능력을 효소액일정량에 대한 분해된 malathion milligram으로 비교하면 관옥>IR-262>후지사까 5호>CP-slo>팔달>수원 82호의 순으로 관옥이 가장 활성이 강하고 팔달, 수원 82호가 가장 약하다. 2. Esterase zymogram을 보면 품종간에 대차없이 $3{\sim}4$개의 band가 두극으로 움직이며 cathode로 특히 굵고 진한 band가 있고 수원 82호는 다른 품종과 약간 다른 pattern을 보였다. 3. p-Nitrophenyl acetate를 기질로 할 때 매 milligram 단백질당의 흡광도로 활성을 비교하면 CP-slo>IR-262>팔달>관옥>수원 82호>후지사까 5호의 순으로 indica type의 품종이 훨씬 활성이 강하나 malathion 경우와 일치하지는 않는다. 4. 0.2ppm 정도의 malathion으로 벼의 esterase는 별로 저해 되지 않았다. 5. 발아중인 벼종자에는 malathion과 p-nitrophenyl acetate를 가수분해하는 복합 esterase가 존재 할 것으로 추측된다.

• Applied Biological Chemistry
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• 제34권1호
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• pp.54-60
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• 1991

식물 병원성 사상균 세포벽의 주성분인 chitin을 분해하는 미생물을 토양에서 분리, 선발하여 동정하고 이들의 효소적 성질을 조사하였다. 선발균 Serratia (S) marcescens는 식물 병원성 사상균인 Fusarium (F) axysporum과 Rhizoctomia (R) solani에 대해 길항력은 갖고 있었으며 효소적으로 chitinase 외에 laminarinase 및 protease 등의 활성을 가지고 있었다. 선발 균주의 chitinase 생산 최적조건은 chitin broth 기본배지의 조성과 온도 및 배양시간을 변형하여 조사한 결과 colloidal chitin 1.5%, tryptone 0.5%, glucose 1%, peptone 0.2%, $MgSO_4{\cdot}7H_2O\;0.1%,\;K_2HPO_4\;0.1%,\;NaCl\;0.1%$ (w/v), pH 6.8 배지에서 $30^{\circ}C$, 72시간 배양하였을 때이었으며 효소의 in vitro 최적 활성조건은 pH 7.5, $50^{\circ}C$이었다. 한편 여러 무기이온 중에서 $Ag^+$와 $Mn^{++}$은 효소의 활성을 촉진시켰다.

• BMB Reports
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• 제42권9호
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• pp.552-560
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• 2009

Cyclooxygenases (COX-1 and COX-2) are ER-resident proteins that catalyze the committed step in prostanoid synthesis. COX-1 is constitutively expressed in many mammalian cells, whereas COX-2 is usually expressed inducibly and transiently. Abnormal expression of COX-2 has been implicated in the pathogenesis of chronic inflammation and various cancers; therefore, it is subject to tight and complex regulation. Differences in regulation of the COX enzymes at the posttranscriptional and posttranslational levels also contribute significantly to their distinct patterns of expression. Rapid degradation of COX-2 mRNA has been attributed to AU-rich elements (AREs) at its 3’UTR. Recently, microRNAs that can selectively repress COX-2 protein synthesis have been identified. The mature forms of these COX proteins are very similar in structure except that COX-2 has a unique 19-amino acid (19-aa) segment located near the C-terminus. This C-terminal 19-aa cassette plays an important role in mediation of the entry of COX-2 into the ER-associated degradation (ERAD) system, which transports ER proteins to the cytoplasm for degradation by the 26S proteasome. A second pathway for COX-2 protein degradation is initiated after the enzyme undergoes suicide inactivation following cyclooxygenase catalysis. Here, we discuss these molecular determinants of COX-2 expression in detail.

• Journal of Microbiology and Biotechnology
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• 제21권1호
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• pp.14-19
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• 2011

A cyclic undecapeptide-family natural product, cyclosporin A (CyA), which is one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex in a filamentous fungal strain named Tolypocladium niveum. Previously, structural modifications of cyclosporins such as a regionspecific hydroxylation at the $4^{th}$ N-methyl leucine in a rare actinomycetes called Sebekia benihana were reported to lead to dramatic changes in their bioactive spectra. However, the reason behind this change could not be determined since a system to genetically manipulate S. benihana has not yet been developed. To address this limitation, in this study, we utilized the most commonly practiced gene manipulation techniques including conjugation-based foreign gene transfer-and-expression as well as targeted gene disruption to genetically manipulate S. benihana. Using these optimized genetic manipulation systems, a putative cytochrome P450 hydroxylase (CYP) gene named CYP506, which is involved in CyA hydroxylation in S. benihana, was specifically disrupted and genetically complemented. The S. benihana${\Delta}$CYP506 exhibited a significantly reduced CyA hydroxylation yield as well as considerable yield restoration by functional complementation of the S. benihana CYP506 gene, suggesting that the genetically manipulated S. benihana CYP mutant strains may serve as a more efficient bioconversion host for various valuable metabolites including CyA.

• Bulletin of the Korean Chemical Society
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• 제33권9호
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• pp.3048-3054
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• 2012

The use of aspirin is widely recommended for the prevention of heart attacks owing to its ability to inhibit platelet activation by irreversibly blocking cyclooxygenase 1. However, aspirin also affects the fibrinolytic and hemostatic pathways by mechanisms that are not well understood, causing severe hemorrhagic complications. Here, we investigated the ability of aspirin and aspirin metabolites to inhibit thrombin-activatable fibrinolysis inhibitor (TAFI), the major inhibitor of plasma fibrinolysis. TAFI is activated via proteolytic cleavage by the thrombin-thrombomodulin complex to TAFIa, a carboxypeptidase B-like enzyme. TAFIa modulates fibrinolysis by removing the C-terminal arginine and lysine residues from partially degraded fibrin, which in turn inhibits the binding of plasminogen to fibrin clots. Aspirin and its major metabolites, salicylic acid, gentisic acid, and salicyluric acid, inhibit TAFIa carboxypeptidase activity. Salicyluric acid effectively blocks activation of TAFI by thrombin-thrombomodulin; however, salicylates do not inhibit carboxypeptidase N or pancreatic carboxypeptidase B. Aspirin and other salicylates accelerated the dissolution of fibrin clots and reduced thrombus formation in an in vitro model of fibrinolysis. Inhibition of TAFI represents a novel hemostatic mechanism that contributes to aspirin's therapy-associated antithrombotic activity and hemorrhagic complications.

• Bulletin of the Korean Chemical Society
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• 제35권9호
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• pp.2803-2808
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• 2014

High potential and fast electron transfer of a cathode mediator are significant factors for improving the performance of biofuel cells. This paper reports the first synthesis of a cathode redox polymer that is a coordination complex of poly (acrylic acid-vinylpyridine-acryl amide) (PAA-PVP-PAA) and [Os(4,4'-dicarboxylic acid-2,2'-bipyridine)$_2Cl_2]^{/+}$ ($E^{\circ}=0.48V$ versus Ag/AgCl). Bilirubin oxidase can be easily incorporated into this polymer matrix, which carried out the four-electron oxygen under typical physiological conditions (pH 7.2, 0.14 M NaCl, and $37^{\circ}C$). This new polymer showed an approximately 0.1 V higher redox potential than existing cathode mediators such as PAA-PVI-$[Os(dCl-bpy)_2Cl]^{+/2+}$. In addition, we suggest increasing the polymer solubility with two hydrophilic groups present in the polymer skeleton to further improve fast electron transfer within the active sites of the enzyme. The maximum power density achieved was 60% higher than that of PAA-PVI-$[Os(dCl-bpy)_2Cl]^{+/2+}$. Furthermore, high current density and electrode stability were confirmed for this osmium polymer, which makes it a promising candidate for high-efficiency biofuel cells.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.354-359
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• 1997

Optimal cultivation conditions for the production of phenylalanine ammonia-lyase(PAL) from Rhodotorula aurantiaca K-505 were selected, and the kinetic parameters of the produced PAL were determined. The most suitable carbon and nitrogen sources were glucose and tryptone, respectively. The strain expressed PAL constituttively when using the optimized semi-complex media. High cell density culture could be critical for maximal production of PAl since the PAL ynthesis was growth associated. maximum PAL activity was observed at initial pH 6.0. although the ll growth was not markedly affected by temperature between 22 and 28$^{\circ}C$, the cells yielded the maximum PAL activity when cultivated at 22$^{\circ}C$. The maximum activity for deamination of L-phenylalnine to trans-cinnamic acid was observed around pH 8.8. The PAL activity gave the maximum at 45$^{\circ}C$, and greatly decreased at higher than 5$0^{\circ}C$. Activation energy({TEX}$E_{a}${/TEX}) calculated from Arrhenius equation was 6.28 kcal/mol in the range of 22$^{\circ}C$ to 4$0^{\circ}C$. A oolf plot showed that the enzyme reaction follows Michaelis-Menten equation, whose {TEX}$K_{M}${/TEX} and {TEX}$V_{max}${/TEX} values were 4.65$\times${TEX}$10^{-3}${/TEX} M and 0.89$\mu$ mol/mg-min respectively.