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한범구(Beom Ku Han),이우진(Woo Jin Lee),유탁(Tak You),박인호(In Ho Park),조도현(Do Hyun Jo) 한국응용생명화학회 1996 Applied Biological Chemistry (Appl Biol Chem) Vol.39 No.6
The survey on the chitinolytic activity of some plants was performed for the purpose of obtaining some reliable and inexpensive sources of chitinase. Rice, soybean for sprouting, kiwi fruit, almond and crude papain were investigated. Rice bran, seed coat of the soybean and the pericarp of kiwi fruit showed a considerable activity, while the bean after the removal of the seed coat, the mixture of rice integument and endosperm, polished rice, and defatted soybean powder didn`t have any detectable activity. These crude enzymes have shown to contain both endo- and exochitinase activity. The effects of pH and temperature on the enzyme activity were variable. Furthermore we have observed the chitosanolytic activity from these enzyme preparations. The rice bran had the highest activity in the enzymatic degradation of chitosan, and seed coat of soybean and the pericarp of kiwi fruit followed. On the basis of the fact that crude papain was not only commercially available but also the most potent in the endochitinase activity and the lowest in the exochitinase activity, we could conclude that crude papain was considered as the most suitable source of the chitinase among plants studied in this paper. In addition, rice bran was worth further investigation from the point of utilizing agricultural by-product.
쥐 뇌의 progesterone 대사에 미치는 연령의 효과
한범구(Beom Ku Han),박인호(In Ho Park),조도현(Do Hyun Jo) 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.1
The effect of age on the metabolism of progesterone was studied in the rat brain. Metabolic activity was more active in minced tissues than total homogenates. The activity of progesterone 5α-reductase(s) was increased during postnatal periods(between 5 and 14 days after birth) and thereafter steadily decreased up to the one-fourth level of the fetus. When 5α-dihydroprogesterone was incubated with brain tissues of various ages, the change in the activity of 3α-hydroxysteroid oxidoreductase(3α-HSOR) was similar to that of 5α-reductase(s). These results suggest that the reduced formation of total 5α-reduced metabolites was due to the decreased activities of 5α-reductase(s) and 3α-HSOR. However the level of 3β-HSOR remained constant regardless of the age.
한범구,이우진,조도현 ( Beom Ku Han,Woo Jin Lee,Do Hyun Jo ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.2
This study was aimed to survey inexpensive and reliable sources of chitinase from the animal origin. The stomach and its content of the broiler, the cod, the yellowtail and β-glucuronidase from snail gut showed a considerable chitinolytic activity, while those of the bas didn`t have any detectable activity. These crude enzymes was found to have both endo- and exochitinase activity. The effects of pH and temperature on the enzyme activity were variable. The hydrolytic products of colloidal chitin by the enzyme preparation from the broiler and the cod were chitooligomers having the degree of polymerization between 3 and 5. Furthermore we observed the chitosanolytic activity from these enzymes. In the degradation of chitosan the thyme of the broiler had the highest activity and β-glucuronidase from snail gut followed. On the basis of the fact that the by-product of the broiler was not only commercially available but also the most potent in the endochitinase activity and the lowest in the exochitinase activity, we conclude that the gizzard and its thyme are considered as the most suitable source of the industrial chitinase among animals studied in this paper.
쥐 뇌의 ${\Delta}^{5}-3{\beta}$-Hydroxy Steroid Acyl 전이효소의 용해 및 Liposome에서의 재구성
고규정,박인호,한범구,조도현,Ko, Kyu-Jung,Park, In-Ho,Han, Beom-Ku,Jo, Do-Hyun 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.4
쥐 뇌의 microsome으로 부터 ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl 전이효소(${\Delta}^{5}-3{\beta}$-OH-SAT)를 detergent를 이용하여 용해한 뒤에 인산지방질 liposome에서 재구성하여 처리하지 않은 microsome의 ${\Delta}^{5}-3{\beta}$-OH-SAT의 활성도와 비교하였다. 용해에 사용한 detergent중에는 deoxycholic acid가 Tween 80이나 Triton X-100 보다 우수한 ${\Delta}^{5}-3{\beta}$-OH-SAT 활성도를 나타냈으며, liposome 생성에 사용한 인산지방질중에서는 난황 및 대두 phosphatidyl-choline이 가장 높은 활성도를 보였으며 그 다음이 phosphatidylethanolamine이었고 phosphatidylserine과 phosphatidic acid는 재구성하지 않은 용해된 효소보다 ${\Delta}^{5}-3{\beta}$-OH-SAT 활성도가 낮았다. 따라서 인산지방질의 head group의 4차 암모늄기나 amine기는 ${\Delta}^{5}-3{\beta}$-OH-SAT의 재구성활성을 촉진시키고 COOH기가 amine기와 함께 존재하거나 amine기가 없으면 이 효소의 활성이 저해됨을 알 수 있었다. Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.
쥐 뇌의 Δ5 - 3β - Hydroxy Steroid Acyl 전이효소의 용해 및 Liposome 에서의 재구성
고규정(Kyu Jung Ko),박인호(In Ho Park),한범구(Beom Ku Han),조도현(Do Hyun Jo) 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.4
Solubilization of microsomal △^5-3β-hydroxy steroid acyl transfearse(△^5-3β-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of △^5-3β-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of △^5-3β-OH-SAT and phosphatidytethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of △^5-3β-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the △^5-3β-OH SAT.
키토산의 효소분해산물로부터 수용성 올리고당의 분리 및 정제
한범구,이우진,조도현 한국키틴키토산학회 1998 한국키틴키토산학회지 Vol.3 No.3
Chitosan with 80% deaceylation and 30cps of viscosity was treated with papain, commercial chitosanase from Bacillus sp.(B-chitosanase), and a chitosanase from the digestive tract of broiler(G-chitosanase). Water-soluble chitosan oligomers were separated from the reaction mixture by differential precipitation and further analyzed by gel filteration. The conversion ratio of chitosan into water-soluble chitosan oligomers was 26% for B-chitosanase, 33% for G-chitosanase, and 39% for papain. With gel filteration of the separated oligomers, the major constituent was identified as dimer of glucosamine for B-chitosanase and papain and the trimer for G-chitosanase.
한범구,조성준,문종국,김새봄,조도현 한국키틴키토산학회 2000 한국키틴키토산학회지 Vol.5 No.3
Low-cost enzyme sources are requested for the production of chitosan oligomers, which were recently spotlighted as functional oligosaccharides. Gizzard tissue of broiler and its chyme are selected for the production of chitosan oligomers, and chitosanolytic activities were investigated. Chitosanolytic activity of chyme was about two times higher than that of gizzard tissue. Repeated freezing and thawing did not affect the activity of crude enzyme preparations. Optimal pH for the gizzard and the chyme were 3.0 and 2.8, respectively. Optimal temperature of the crude enzymes ranged from 60℃ to 70℃ and their activities were maintained at 70% level at 80℃. At 50℃ for three days no decrease in their activity was observed. After 24 hour incubation, gizzard and chyme produced 40 yg oligomers/mg-protein and 80 yg oligomers/mg-protein, respectively. Under optimal conditions, 62% of total substrate were made water-soluble by the gizzard. Trimer of glucosamine was identified as its main reaction product.