• 한국버섯학회지
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• 제19권4호
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• pp.316-321
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• 2021

리그닌 분해효소의 생산은 나선형 리본이 있는 새로운 형태의 회전식 통풍관 생물 반응기(RTB)를 사용하여 느타리(Pleurotus ostreatus) No.42에 의해 실시하였다. 락게이즈(laccase)의 최대 생산량은 배양 3일 후 약 8,200 U/bioreactor 수준에 도달한 후 감소하였다. 반면에, 망간퍼옥시데이즈(MnP)의 최대 생산은 6일 배양 후 약 8,400 U/bioreactor의 수준에 도달하였다. 그러나 이 발효조에서 리그닌퍼옥시데이즈(LiP)는 검출되지 않았다. 그 결과 회전식 통풍관 생물 반응기(RTB)가 리그닌 분해효소를 대규모 생산을 위해 성공적으로 생산할 수 있음을 보여주었다. 이 발효기에서 망간퍼옥시데이즈의 정제 과정은 Sepharose CL-6B, Superdex 75 prep grade 및 Mono-Q에 대한 크로마토그래피를 포함하여 정제하였다. 이 주요 효소는 sodium dodecyl sulfate-polyacrylamide겔 전기영동(SDS-PAGE)에서 분자량 36,400, pI 3.95의 등전점(IEF)으로 각각 확인되었다. 이 발효기의 주요 효소 N-말단 서열은 정치 및 진탕배양과 같은 다른 배양조건에서 보고된 MnP3 효소와 동일하였다.

• 한국환경성돌연변이발암원학회지
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• 제11권2호
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• pp.148-160
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• 1991

인체의 여리 조직을 대상으로하여 면연조직화학전방법을 통하여 glutathione transferase isoenzyme의 분포를 비교 분석하였다. 상피조직중의 GST-Pi의 다랑분포가 확인되었으며, 특히 외인성 화학물질에 노출되기 쉬운 피부, 유두, 신도 등의 과립층과 췌장, 담도, 타액선, 신세뇨관등의 배설관의 상피 또는 난소의 theca와 granulosa, 고환의 Leydig세포, 부신피 질의 zona reticularis 등의 steroid 생합성이 왕성한 부위에서 높은 GST-Pi 분포가 관찰되었다. 면역조직화학적 분석과 병행하여 Western blot 분석을 시행해본 결과 이러한 GST-Pi 분포가 인체유래 유각상피세포 중에서도 확인되었다. 그러나 K-ras 암유전자로 형질전환하여 암화능을 갖춘 인체상피세포주의 GST-Pi 발현은 유의한 차이를 보이지 않았다. 뿐만아니라 타액중에서도 GST-Pi가 다량 존재함이 Western blot 분석에 의하여 구명되었다. 이러한 결과는 GST-Pi가 인체조직 중 외인성독성물질에 노출되는 부위, 내인성 steroid 합성부위 및 배설능을 갖는 부위의 상피조직에 주로 존재하고 있음을 보여주고 있으며, 이러한 GST-Pi의 조직분포는 동효소가 인체의 해독기능에 주요한 역할을 하고 있음을 시사해주고 있다.

• Parasites, Hosts and Diseases
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• 제34권4호
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• pp.259-266
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• 1996

형태학적 제3군 가시아메바의 taxonomic validity는 아직 확실하지 않다. 이번 연구에서 제3군에 속하는 6종의 가시 아메바 즉 A. culbertsoni A. healyi A. palestinensis. A. pustulosa, A. royreba 및 A. lenticulata의 type strain들의 동위효소. 미토콘트리아 DNA 및 small subunit(ssu) rDNA의 Restriction Fragment Length Polymorphism(RFLP) 양상을 비교하여 이들의 taxonomic validity를 검토하였다. 미토콘드리아 DNA의 RFLP 양상은 분리주간에 서로 심한 차이를 보였다. A. palestinensis와 A. pustulosc는 거의 동일한 rDNA RFLP(추정 염기 치환율 2.6%) 및 동위효소의 양상을 보여 A. palestinensis와 A. pustulosc는 같은 종으로 판단되었다. 그외의 종들은 서로 아주 다양한 rDNA RFLP 및 동위효소의 양상을 나타내어 독립종으로 인정되었다.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1570-1578
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• 2016

Commercial application of laccase is often hampered by insufficient enzyme stocks, with very low yields obtained from natural sources. This study aimed to improve laccase production by mutation of a Coriolopsis gallica strain and to determine the biological properties of the mutant. The high-yield laccase strain C. gallica TCK was treated with N-methyl-N-nitro-N-nitrosoguanidine and ultraviolet light. Among the mutants isolated, T906 was found to be a high-production strain of laccases. The mutant strain T906 was stabilized via dozens of passages, and the selected ones were further processed for optimization of metallic ion, inducers, and nutritional requirements, which resulted in the optimized liquid fermentation medium MF9. The incubation temperature and pH were optimized to be 30℃ and 4.5, respectively. The mutant strain T906 showed 3-times higher laccase activity than the original strain TCK under optimized conditions, and the maximum laccase production (303 U/ml) was accomplished after 13 days. The extracellular laccase isoenzyme 1 was purified and characterized from the two strains, respectively, and their cDNA sequence was determined. Of note, the laccase isoenzyme 1 transcription levels were overtly increased in T906 mycelia compared with values obtained for strain TCK. These findings provide a basis for C. gallica modification for the production of high laccase amounts.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2571-2576
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• 2012

Breast cancer is the first of the most common ten cancers in Iraq. Its etiology is multifactorial, oxidative stress and lipid peroxidation being suggested to play important roles in carcinogenesis. The purpose of this study was to investigate the oxidant-antioxidant status in breast cancer patients, by measuring SOD isoenzyme activities (total SOD, CuZn-SOD, Mn-SOD and EC-SOD) in plasma and breast tumors, and by estimating thiobarbituric reactive substances (TBRS) in tissue homogenates. General increase in total SOD activity was observed in plasma and tissue samples of breast tumors, greater in the malignant when compared to benign group (p<0.05). Mn-SOD showed a significant decrease in tissue malignant samples (p<0.05), and insignificant decrease in plasma malignant samples compared with control and benign samples. Plasma EC-SOD activity in both patient benign and malignant breast tumors demonstrated 3.5% and 22.8% increase, respectively. However, there was a decrease in tissue EC-SOD activity in malignant breast tumors when compared with benign. A similar tendency was noted for TBRS. We suggest that elevated total SOD might reflect a response to oxidative stress, and then may predict a state of excess reactive oxygen species in the carcinogenesis process. If there is proteolytic removal of the heparin binding domain, EC-SOD will lose its affinity for the extracellular matrix and diffuse out of the tissue. This will result in a decreased EC-SOD activity, thus leading to an increase in the steady-state concentration of $O^{2-}$ in this domain, and increase in EC-SOD activity in the extracellular fluid. This might explain the results recorded here concerning the decrease in tissue EC-SOD activity and increase in plasma of breast cancer patients.

• 한국약용작물학회지
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• 제24권1호
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• pp.7-13
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• 2016

Background : Ginger has been extensively used in foods and traditional medicines in Asian countries. Despite its frequent consumption in daily life, the mechanism of potential interactions between ginger components-drug has not been examined. To elucidate the mechanism of governing the effects of 6-shogaol, a primary constituent of dried ginger, on human cytochrome P450 (CYP) isoenzymes an incubation studies were carried out using pooled human liver microsome (HLM). Methods and Results : CYP isoenzyme specific substrate was incubated with multiple concentrations of inhibitor, HLM and cofactors. 6-shogaol showed a potent inhibitory effect on CYP2C9, CYP1A2 and CYP2C19 with half maximal inhibitory concentration ($IC_{50}$) values of 29.20, 20.68 and $18.78{\mu}M$ respectively. To estimate the value of the inhibition constant ($K_i$) and the mode of inhibition, an incubation study with varying concentrations of each CYP isoenzyme-specific probe was performed. 6-shogaol inhibited CYP2C9 and CYP2C19 noncompetitively ($K_i=29.02$ and $19.26{\mu}M$ respectively), in contrast, the inhibition of CYP1A2 was best explained by competitive inhibition ($K_i=6.33{\mu}M$). Conclusions : These findings suggest that 6-shogaol may possess inhibitory effects on metabolic activities mediated by CYP1A2, CYP2C9 and CYP2C19 in humans.

• 한국미생물·생명공학회지
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• 제16권4호
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• pp.259-264
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• 1988

Pseudomonas sp.로 동정된 토양분리균의 inulinase를 ammonium sulfate 분획, DEAE Sephadex A-50 Chromatography, Sephadex G 100 및 Sephadex G200 gel여과 등의 과정을 거쳐 정제 한 결과 inulinase PI과 PII의 두 isoenzyme으로 분리되었으며 각각 약 24배와 17배 정제되어 단일단백질로 분리되었다. PI, PII 두 isoenzyme은 다같이 sucrose, raffinose 및 levan은 분해하지 못하고 inulin만을 endo-type로 분해 절단하는 $\beta$-2,1-fructanfructano hydrolase(EC 3.2.1.7)임이 밝혀졌다. 효소활성 최적온도는 두 효소가 같은데 비해 (55$^{\circ}C$), 최적 pH는 PI이 pH5.5, PII가 pH6.0으로 약간의 차이를 보였다. Inulin에 대한 Km값은 PI ; 2$\times$$10^{-3}$M, PII : 5$\times$$10^{-3}$M로써 PI효소가 PII보다 약간 높은 친화력을 보였다.

• Parasites, Hosts and Diseases
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• 제37권2호
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• pp.85-92
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• 1999

To determine the pathogenicity of Acanthamoeba spp. isolated in Korea and to develop a isoenzymatic maker, the mortality rate of infected mice, in vitro cytotoxicity against target cells and isoenzyme band pattern were observed. Five isolates of Acanthamoeba spp. (YM-2, YM-3, YM-4, YM-5 and YM-7) were used in this study as well as three reference Acanthamoeba spp. (A. culbertsoni, A. hatchetti, and A. royreba). According to the mortality rate of infected mice, Korean isolated could be categorized into three groups: high virulent (YM-4), low virulent (YM-2, YM-5, YM-7) and the nonpathogenic group (YM-4), In addition, the virulence of Acanthamoeba spp. was enhanced by brain passage in mice. In the cytotoxicity assay against chinese hamster ovary cells, especially, the cytotoxicity of brain-passaged amoebae was relatively higher than the long-term cultivated ones. The zymodeme patterns of glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), hexokinase (HK), glutamate oxaloacetate transaminase (GOT) and malic enzyme (ME)of Acanthamoeba spp. were different among each isolate, and also between long-term cultrued amoebae and brain passaged ones. In spites of the polymorphic zymodemes, a slow band of G6PD and K, and an intermediate band of MDH were only observed in pathogenic Acanthamoeba spp., which should be used as isoenzymatic makers.

• Journal of Chest Surgery
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• 제26권6호
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• pp.441-451
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• 1993

Protective effect of superoxide dismutase[SOD] and substrates on acute ischemic and reperfused myocardium was assessed by cardiac microdialysis. 30 Rabbits were divided into 4 groups; normal control group [group I, n=5], ischemic group [group II, n=5], SOD treated group [group III, n=10], and substrates treated group [group IV, n=10]. After a microdialysis apparatus was implanted in rabbit myocardium, coronary artery was occuluded for 5 minutes and reperfusion was performed for 30 minutes. Hemodynamic changes, CK-MB isoenzyme level and adenine ring compound level in effluent dialysates [equilibrated with interstitial fluid], and ultrastructural changes of myocardial cell were analysed. Systolic blood pressure at 10 and 30 minutes after reperfusion was higher in group III and IV than in group II [p<.05]. Also percent recovery of systolic blood pressure in group III [p<.01] and IV [p<.02] was higher than in group II. CK-MB isoenzyme level in effluent dialysates was peaked at 10 minutes after reperfusion, thereafter decreased in group II, III and IV. At 30 minutes after reperfusion, its level was lower in group III and IV than in group II[p<.05]. Adenine ring compound level in effluent dialysates increased till 10 minutes after reperfusion and progressively decreased. At 10 and 30 minutes after reperfusion, its level was lower in group III and IV than in group II without significance. Degree of myocardial damage was estimated by scoring of mitochondrial injury. Group I was within normal range and most severe injury was seen in group II. And the score of mitochondrial injury in group III and IV was lower than in group II. In conclusion, SOD and substrates[KMP solution] had protective effect on stunned myocardium. The microdialysis appratus was a good device for studying stunned myocardium, and cardiac microdialysis might be a unique technique for analysis of regional intramyocardial interstitial fluid.

• 한국식품영양과학회지
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• 제19권6호
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• pp.596-604
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• 1990

Polygalacturonase의 활성은 미숙감과실에서 나타나지 않았으나 완숙감과 연시에서 각각 55.01. 206.70unit/100g-fr. wt.로 연시에서 급격히 증가하였다. 연시에서 추출한 polygalacturonase의 isoenzyme은 2종이었고, 분자량은 다같이 55,000 dal-ton이었다. Polygalacturonase isoenzyme의 특성을 조사한 결과, polygalacturonase I의 Vmax는 0.195$\mu$ mole reducing-sugar/$m\ell$/30min., Km값은 3.50mg/$m\ell$이었고, 최적온도 4$0^{\circ}C$, 최적 pH 3.5이었으며, 열처리시 6$0^{\circ}C$까지 안정하였다. Polygalacturonase II 의 Vmax는 0.11$\mu$mole reducing-sugar /$m\ell$/30min., Km값은 2.50mg/$m\ell$이었고, 최적 온도와 pH는 35$^{\circ}C$와 4.0이었으며. 4$0^{\circ}C$까지 안정하였다. Polygalacturonase I 과 II는 K+, Cu++, Zn++, SDS, EDTA에 의해서 활성이 저해되었고 Ca++ 의해 다소 촉진되었다.