• Bulletin of the Korean Chemical Society
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• 제33권10호
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• pp.3233-3240
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• 2012

We demonstrated the combined applications of online protein digestion using trypsin immobilized enzyme reactor (IMER) and dual tandem mass spectrometry with collisionally activated dissociation (CAD) and electron transfer dissociation (ETD) for tryptic peptides eluted through the trypsin-IMER. For the trypsin-IMER, the organic and inorganic hybrid monolithic material was used. By employing the trypsin-IMER, the long digestion time could be saved with little or no sacrifice of the digestion efficiency, which was demonstrated for standard protein samples. For three model proteins (cytochrome c, carbonic anhydrase, and bovine serum albumin), the tryptic peptides digested by the IMER were analyzed using LC-MS/MS with the dual application of CAD and ETD. As previously shown by others, the dual application of CAD and ETD increased the sequence coverage in comparison with CAD application only. In particular, ETD was very useful for the analysis of highly-protontated peptide cations, e.g., ${\geq}3+$. The combination approach provided the advantages of both trypsin-IMER and CAD/ETD dual tandem mass spectrometry applications, which are rapid digestion (i.e., 10 min), good digestion efficiency, online coupling of trypsin-IMER and liquid chromatography, and high sequence coverage.

• 한국식품위생안전성학회지
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• 제10권3호
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• pp.163-168
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• 1995

The present study has been undertaken to investigate the effects of garlic added to food on the activities of several enzymes in serum of rats fed lard and alcohol. Thirty-five males of Sprague-Dawley strains weighed about 130g were divided into 7 groups, each group receiving a different diet for 10 weeks; i.e. basal diet plus 15% lard, basal diet plus 5% alcohol, basal diet plus 0.5% garlic, basal diet plus 15% lard and 0.5% garlic. Determinations were carried out in the net weight gain, food efficiency ratio, weight of organs, and AST, ALT, lactate dehydrogenase, alkaline phosphatase activities in serum of rats. The results obtained were as follows; Rats given feed containing lard and alcohol showed significant decrease in net weight gain, but garlic caused an increased in food efficiency ratio. Lard supplementation caused an increase in the weight of liver, kidney, spleen, but another groups did not, AST, ALT, ALP, LDH of serum were significantly increased in lard and alcohol containing group but garlic feeding decreased enzyme activities compared to lard and alcohol containing group. The above results suggest that garlic would prevent the metabolic disease of liver by improving hyperlipemia caused by high fat diet.

• Journal of Nutrition and Health
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• 제34권7호
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• pp.741-747
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• 2001

The effects of fruiting body of Cordyceps militaris on the growth, the food intake, the food efficiency ratio, the lipid metabolism, the serum protein level ad enzyme activity in male rats were studied. Sprague-Dawley rats were fed four types of diets for five weeks, respectiely: a control diet, a control diet supplemented with 2%, 3% or 4% fruiting body of Cordyceps militaris(CF) powder. In rats fed 2% or 3% CF diets the body weight gain, the food intake, the concentratons of hepatic triglyceride and serum LDL-cholesterol, the atherogenic index, and the total lipid, total cholesterol, triglyceride and phospholipid in serum were similar to those in rats fed the control diet. Whereas, in 4% CF diet these were significantly decrased. But the all CF diets feeding could not decrease the food efficiency ratio, the weights of liver, pancreas, spleen, kidney and heart, and the concentration of serum HDL-cholesterol. Also, it was shown that the concentrations of glucose, total protein, albumin, urea and creatinine, and the GOT, GPT, LDH, ${\gamma}$-GTP and ALP activities were the same levels in serum of rats of fed all the experimental diets.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.633-634
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• 2001

Porcine pancrease lipase를 aldehyde-silica에 in situ 상태에서 immobilization 할때의 최적 조건에 대하여 연구하였다. Feed 용액의 조성비율, 양, pH, 온도, 농도, 유속에 대하여 최적화하였으며 lipase의 activity를 측정 비교하였다.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.205-210
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• 2013

Thiol-ene polymerization is a versatile tool for several applications. Here we report the preparation of epoxide groups containing thiol-ene photocurable polymeric support and the covalent immobilization of ${\alpha}$-amylase onto these polymeric materials. The morphology of the polymeric support was characterized by scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) coupled with SEM was used to explore the chemical composition. The polymeric support and the immobilization of the enzyme were characterized by FTIR analysis. SEM-EDS and FTIR results showed that the enzyme was successfully covalently attached to the polymeric support. The immobilization efficiency and enzyme activity of ${\alpha}$-amylase were examined at various pH (5.0-8.0) and temperature ($30-80^{\circ}C$) values. The storage stability and reusability of immobilized ${\alpha}$-amylase were investigated. The immobilization yield was $276{\pm}1.6$ mg per gram of polymeric support. Enzyme assays demonstrated that the immobilized enzyme exhibited better thermostability than the free one. The storage stability and reusability were improved by the immobilization on this enzyme support. Free enzyme lost its activity completely within 15 days. On the other hand, the immobilized enzyme retained 86.7% of its activity after 30 days. These results confirm that ${\alpha}$-amylase was successfully immobilized and gained a more stable character compared with the free one.

• 한국군사과학기술학회지
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• 제2권2호
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• pp.140-147
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• 1999

A gene expressing organophosphorus acid (OPA) anhydrolases have been cloned from Alteromonas haloplanktis strain and expressed in bacterial strain BL21. Crude extract was prepared from the transformed bacterial strain BL2l and used in testing its degrading capability of real nerve gas, soman and sarin. Within 1 minute after the start of the reaction, nearly 65% of the soman added to the reactant(3mM) was degraded by adding 1 mg of the crude extract enzyme(20.0 Unit $mg^-{1}$ crude protein). In 6 minutes, the reaction reached at its steady state, which indicates that soman was completely degraded by that time. In the case of sarin, the degradation efficiency was observed to be about 0.7 times of that of soman. If the specific activity of OPAA is enhanced by both increased expression efficiency and purification, OPAA seems to be applied for the development of decontaminant of skin, especially of eye.

• 한국미생물·생명공학회지
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• 제20권1호
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• pp.40-45
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• 1992

Aspergillus niger로부터 acidic nucleotidase를 Sepharose CL-6B gel 여과와 DEAE-Sephacel 이온교환수지를 이용하여 부분정제하였다. 5'-AMP 와 3'-AMP를 기질로 사용했을 경우에 효소의 최적 pH는 4.5, 그리고 최적온도는 $55^{\circ}C$였다. 그러나, p-nitrophenyl phosphate를 기질로 사용했을 경우에는 최적 pH는 변화가 없었으나, 최적 온도는 $70^{\circ}C$였다. 효소의 활성화에너지는 3'-AMP, 5'-AMP 그리고 p-nitrophenyl phosphate를 기질로 사용했을 경우에 각각 4.76kcal/mole, 6.95kcal/mole 그리고 11.82kcal/mole 였다.

• Journal of Microbiology and Biotechnology
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• 제2권1호
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• pp.14-20
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• 1992

Some microorganisms isolated from soil, including some bacteria and fungi, were found to secrete an extracellular soymilk-clotting enzyme. Among them, an isolated fungus showed the highest soymilk-clotting activity and the strain was assigned to genus Penicillium based on its cultural and morphological characteristics, and designated as Penicillium sp. L-151K. Soymilk-clotting enzymes A and B produced by Penicillium sp. L-151K were purified by ammonium sulfate precipitation and chromatographies on Sephadex G-25, CM-Sephadex, Sephadex G-100 and phenyl-Toyopearl gel. The two purified enzymes A and B were found to be homogeneous by polyacrylamide gel electrophoresis at pH 9.5. The molecular weights of enzyme A and B were 24, 000 and 40, 000, respectively, by gel filtration on Sephadex G-100. Enzymes A and B coagulated soymilk optimally at $60^\circ{C}$ and were stable up to $50^\circ{C}$. Both enzymes were most active at pH 5.8 for soymilk coagulation, and were stable with approximately 80% of original activity from pH 3.0 to 5.0. Each enzyme was an acidic protease with an optimum pH of 3.0 for casein digestion. The soymilk-clotting efficiency of these enzymes was improved with $CaCl_2\;or\;MgCl_2$ when making soymilk-curd.

• 한국환경보건학회지
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• 제31권4호
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• pp.294-300
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• 2005

Food waste can be a valuable carbon source in biological nutrient removal (BNR) systems because of high C/N and C/P ratios. However, food waste should be pretreated to promote its hydrolysis rate because hydrolysis reaction would be a rate-limiting step. This study investigates the influence of the enzymatic pretreatment on acid fermentation of food waste. Solubilization of particulate matter in food waste by using commercial enzymes was examined. The acidification efficiency and the volatile fatty acids (VFAs) production potential of enzymatically pretreated food waste were also examined. The highest volatile suspended solids (VSS) reduction was obtained with an enzyme mixture ratio of 1:2:1 of carbohydrase:protease:lipase. An optimum enzyme dosage for solubilization of food waste was $0.1\%$(V/V) with the enzyme mixture ratio of 1:2:1. In the acid fermentation of enzymatically pretreated food waste, $0.1\%$(V/V) enzyme mixture dosage for pretreatment result in the maximum VFAs production and the best VFAs fraction in soluble COD(SCOD). The VFAs production at this addition level was 3.3 times higher than that of no-enzyme added fermenter. The dominant VFAs present was n-butyrate followed by acetate.

• Journal of Microbiology and Biotechnology
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• 제21권7호
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• pp.711-718
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• 2011

A highly efficient cellobiohydrolase (CBH)-secreting basidiomycetous fungus, Agaricus arvensis KMJ623, was isolated and identified based on its morphological features and sequence analysis of internal transcribed spacer rDNA. An extracellular CBH was purified to homogeneity from A. arvencis culture supernatant using sequential chromatography. The relative molecular mass of A. arvencis CBH was determined to be 65 kDa by SDSPAGE and 130 kDa by size-exclusion chromatography, indicating that the enzyme is a dimer. A. arvencis CBH showed a catalytic efficiency ($k_{cat}/K_m$) of 31.8 $mM^{-1}\;s^{-1}$ for p-nitrophenyl-${\beta}$-D-cellobioside, the highest level seen for CBH-producing microorganisms. Its internal amino acid sequences showed significant homology with CBHs from glycoside hydrolase family 7. Although CBHs have been purified and characterized from other sources, A. arvencis CBH is distinguished from other CBHs by its high catalytic efficiency.