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소의 뇌에서 Neurosteroid Acyltransferase의 분포 및 특성에 관한 연구
박인호,조성준,조도현 ( In Ho Park,Sung Jun Jo,Do Hyun Jo ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.2
The enzymatic properties as well as its distribution in the cerebral region and subcellular organells were investigated for the neurosteroid acyltransferase from the bovine brain, which synthesize the fatty acid esters of the neurosteroids. The cerebellum region was the highest in NSAT activity while the cerebrum was the lowest with 50% of the cerebellar activity. The NSAT was found to be mainly localized in the microsomal fraction. The optimal temperature and pH were 40℃ and 4.9, respectively. When ³H-DHEA was utilized as substrate, the K_m and V_(max) was 32.6 μM and 4.86 nmole/㎎ protein/h, respectively. Under the same condition pregnenolone(△^5P) was a competitive inhibitor with K₁=22.8 μM and testosterone was a uncompetitive inhibitor with K₁=28.2 μM. This may suggest that the NSAT has a different conformation in the acylation of the β-hydroxyl group at C-3 and C-17.
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.
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(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.
최민호,조도현,박연희 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.4
젖소목장의 토양, 돼지 비게, 치즈 등으로부터 cholesterol을 유일 탄소원으로 이용하여 생육하는 박테리아 6균주를 분리하였다. 이 균주들의 형태학적, 생화학적, 생리학적 특성을 조사하여 이 중 3균주는 Rhodococcus species로 부분 동정하고, 각각 Rhodococcus sp. CD-1, CD-2, CD-3으로 명명하였다. 각 균주의 cholesterol 분해능력을 30℃에서 배양하면서 조사한 결과 Rhodococcus spp. CD-1, CD-2, CD-3는 3일 후에 거의 100%를 분해하였으며, 모든 균주들이 주요 분해산물로 cholest-4-en-3-one을 형성하였다. 그러나, cholest-4-en-3-one 이후의 분해단계에서는 균주에 따라 대사방식에 차이가 있어서, Rhodococcus sp. CD-1, CD-3는 cholesterol이 cholest-4-en-3-one으로 산화된 후 모핵의 분해가 이루어지면서 수용성 물질로의 분해가 활발하여 sterol계 대사산물을 거의 남기지 않았으나, Rhodococcus sp. CD-2는 배양 후 cholest-4-en-3-one이 상당량 남아 있어서 cholest-4-en-3-one 이후의 분해단계가 상대적으로 활발하지 못함을 알 수 있었다. Six bacterial strains capable to grow on medium containing cholesterol as sole carbon source were isolated from soil, pork fat and cheese. Three of them were tentatively identified as Rhodococcus species, Rhodococcus sp. CD-1, R. sp. CD-2, and R. sp. CD-3. All the isolates showed a varying amount of cholest-4-en-3-one as the degradation product, and three strains of Rhodococcus spp. showed rapid degradation of cholesterol. Radioisotopic studies revealed that cholesterol was degraded to non-sterol hydrophilic compounds via cholest-4-en-3-one, and presumably to CO_2. These strains showed two distinct patterns in further degradation of cholest-4-en-3-one. By one group, R. sp. CD-1 and R. sp. CD-3, cholest-4-en-3-one was rapidly converted to non-sterol intermediates without significant accumulation of sterol derivatives in the culture broth. In contrast, by another group, R. sp. CD-2, the substantial amount of cholest-4-en-3-one was accumulated indicating a lower conversion of the compound to the next metabolites.
한범구,백제현,조성준,박인호,이우진,조도현 한국키틴키토산학회 1999 한국키틴키토산학회지 Vol.4 No.1
Purification of two chitinases, LMC(low molecular weight chitinase) and HMC(high molecular weight chitinase)from gizzards of broiler was carried out using ion-exchange chromatography Eel-filtration chromatofocusing and hydrophobic interaction chromatography. They were purified about 180 folds with a 3~5% yield. The molecular weight of LMC and HMC were 48.2 kDa and 57.8 kDa, respectively. Both were endotype chitinases having optimum pH of 4~5 and optimum temperature of 60℃. Both enzymes had different Km and Vmax values toward [3H]- caitin. The NH2 -terminal sequences of LMC and HMC shared high level of homology with those reported in animal chitinases.
한범구,유탁,이우진,조성준,박인호,조도현 한국키틴키토산학회 1997 한국키틴키토산학회지 Vol.2 No.3
Crude chitinolytic enzymes were prepared from the chyme, the mucous lining and the remaining tissue of the gizzard, and some enzymatic properties were investigated. Freezing and thawing of the sample did not affect the activity of the subsequent crude preparations of the enzyme. The optima: incubation time and temperature were 2 hours and 10'C, respectively. The pH optima and thermostability were variable, depending upon the source of the enzyme and the type of enzyme action. Under experimental conditions 165mg of N-acetylchitooligomers could be obtained from a gizzard after an incubation time of 18 hours.