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Characterization of a Novel Glutathioe S - Transferase from Pseudomonas sp . DJ77
Uhee Jung,Young Sik Cho,Hark Mo Seong,Seong Jae Kim,Young Chang Kim,An Sik Chung 생화학분자생물학회 1996 BMB Reports Vol.29 No.2
A novel glutathione S-transferase from Pseudomonas sp. DJ77 was expressed in E. coli and purified by glutathione-affinity chromatography. The enzyme was composed of two identical subunits. The molecular size of the enzyme was 42 kDa by sephadex G-150 gel permeation chromatography and Mr of each subunit was 23 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis. pI value of the enzyme was approximately 5.8 by isoelectric focusing. This enzyme showed the highest activity toward 1-chloro-2,4-dinitrobenzene as the electrophilic substrate. The relative activities toward p-nitrobenzyl chloride and 1,2-dichloro-4-nitrobenzene were 3.8% and 1.31 of the activity toward 1-chloro-2,4-dinitrobenzene, respectively. K_m and V_(max) values for 1-chloro-2,4-dinitrobenzene calculated by Lineweaver-Burk plot were 0.76 mM and 14.81 μ㏖/min/㎎, respectively, and those for glutathione were 6.23 mM and 64.93 μ㏖/min/㎎, respectively. The enzyme showed highest glutathione S-transferase activity at pH 8.0 and was stable between pH 6.0 and 9.0. The enzyme retained its activity up to 3590 for 90 min but was unstable above 45℃.
생약조성물(MH-30)의 면역조혈계 및 재생조직 방사선 손상에 대한 방호 효과
정우희(Uhee Jung),박혜란(Hae-Ran Park),이호용(Ho-Yong Lee),백가영(Ga-Young Baek),조성기(Sung-Kee Jo) 한국식품영양과학회 2016 한국식품영양과학회지 Vol.45 No.7
본 연구팀에서는 방사선에 의한 면역조혈계 및 위장관계 손상에 대한 방호를 위하여 당귀, 천궁, 작약 혼합물의 열수추출물로부터 그 다당체 함량을 강화한 생약조성물 Hemo HIM을 개발한 바 있다. 본 연구에서는 열수 추출물에 기반을 두어 제조된 HemoHIM에 비하여 지용성 폴리페놀 성분을 강화함으로써 더 뛰어난 생리활성을 갖는 생약조성물을 개발하고자 30% 에탄올 추출과 열수 추출을 함께 시행하여 얻은 추출물을 기반으로 생약조성물 MH-30을 제조하였다. MH-30과 HemoHIM의 에탄올 분획 내의 성분을 HPLC로 비교 분석한 결과 수용성 성분에는 큰 차이가 없었으나 여러가지 지용성 성분이 MH-30에서 크게 증가하는 것을 확인하였으며, 특히 decursin의 함량은 8.7배로 크게 증가하였다. 다음으로 MH-30의 시험관 내 항산화 및 면역세포 활성화 효과와 방사선 조사 마우스에서 면역조혈계 및 재생조직의 방호 효과를 HemoHIM과 비교하여 관찰하였다. 항산화 활성을 비교하기 위한 시험관 내 hydroxyl radical 및 superoxide anion 소거 활성 평가에서 MH-30이 HemoHIM 보다 높은 항산화 활성을 보였다. 림프구 증식능을 이용한 시험관 내 면역세포 활성화 시험에서는 MH-30과 Hemo HIM은 거의 비슷한 활성을 나타내었다. 방사선 조사 마우스에서 MH-30은 내재성 비장 조혈세포 집락수를 증가시키고 골수조직 내 세포사멸을 줄였으며, 위장관 재생조직인 소장움의 생존율을 증가시키는 등 방사선에 의한 면역조혈계와 재생조직 손상을 방호하는 효과를 보여주었다. 또한, 종합적인 방사선 방호 효과를 평가하는 지표인 방사선 조사 마우스의 30일 생존율도 MH-30 투여로 유의하게 증가함을 확인하였다. 특히 상기한 모든 마우스 실험에서 MH-30은 Hemo HIM보다 더 뛰어난 방호 효과가 있음을 관찰할 수 있었다. 이상의 결과들은 새롭게 개발한 생약조성물 MH-30이 면역조혈계와 재생조직의 방사선 손상을 줄여주는 효과가 있으며 기존에 개발된 HemoHIM에 비해 더 뛰어난 활성을 갖고 있음을 보여주었다. 따라서 MH-30은 방사선 사고 또는 암환자의 방사선치료 시 발생할 수 있는 면역조혈계 및 재생조직의 손상을 경감시킬 수 있는 방사선 방호 물질로서 유용하게 활용될 수 있을 것으로 생각한다. We previously developed an herbal composition (HemoHIM) based on the water extracts of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix to protect and recover hematopoietic and intestinal tissues against radiation injuries. In this study, to develop a composition with improved activities based on enhanced fat-soluble polyphenol contents, we prepared a new herbal composition, MH-30, from the above three herbs by 30% ethanol extraction and hot water extraction. HPLC analysis of the ethanol fractions of MH-30 and HemoHIM revealed that MH-30 had higher contents of many fat-soluble polyphenol compounds than HemoHIM (8.7-fold increase for decursin), whereas contents of water-soluble polyphenol compounds showed little differences between the two compositions. Then, we evaluated MH-30 and HemoHIM for their in vitro antioxidant and immune cell-stimulating activities as well as in vivo protective effects against radiation injuries in hematopoietic and self-renewal tissues. In antioxidant activity assays, MH-30 showed higher hydroxyl radical scavenging activity than HemoHIM (1.4- to 1.9-fold for compositions and 2.3- to 4.5-fold for ethanol fractions). On the other hand, MH-30 and HemoHIM exhibited similar immune cell-stimulating activities as measured by in vitro lymphocyte proliferation. MH-30 increased endogenous spleen colony formation, decreased bone marrow cell apoptosis, and enhanced survival of intestinal crypts in irradiated mice, demonstrating effective protection of MH-30 against radiation-induced injuries in hematopoietic and self-renewal tissues. The 30-day survival rate of lethally irradiated mice, a comprehensive index for radioprotective efficacy, was also elevated by MH-30. Noticeably, MH-30 showed higher protective effects than HemoHIM in all mouse experiments. These results demonstrate that MH-30 can protect hematopoietic and self-renewal tissues against radiation injuries more effectively than HemoHIM. Therefore, MH-30 can be a good candidate to reduce radiation injuries in hematopoietic and self-renewal tissues incurred by radiation accidents or cancer radiation therapy.
Screening of Crude Plant Extracts with Anti-Obesity Activity
Roh, Changhyun,Jung, Uhee Molecular Diversity Preservation International (MD 2012 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.13 No.2
<P>Obesity is a global health problem. It is also known to be a risk factor for the development of metabolic disorders, type 2 diabetes, systemic hypertension, cardiovascular disease, dyslipidemia, and atherosclerosis. In this study, we screened crude extracts from 400 plants to test their anti-obesity activity using porcine pancreatic lipase assay (PPL; triacylglycerol lipase, EC 3.1.1.3) <I>in vitro</I> activity. Among the 400 plants species examined, 44 extracts from plants, showed high anti-lipase activity using 2,4-dinitrophenylbutyrate as a substrate in porcine pancreatic lipase assay. Furthermore, 44 plant extracts were investigated for their inhibition of lipid accumulation in 3T3-L1 cells. Among these 44 extracts examined, crude extracts from 4 natural plant species were active. <I>Salicis Radicis Cortex</I> had the highest fat inhibitory activity, whereas <I>Rubi Fructus</I>, <I>Corni Fructus</I>, and <I>Geranium nepalense</I> exhibited fat inhibitory capacity higher than 30% at 100 μg/mL in 3T3-L1 adipocytes, suggesting anti-obesity activity. These results suggest that four potent plant extracts might be of therapeutic interest with respect to the treatment of obesity.</P>
Kun Ho Chung,Uhee Jung,Kyung-Suk Suh 한국방사성폐기물학회 2023 한국방사성폐기물학회 학술논문요약집 Vol.21 No.2
The radioactive cesium, released from the normal operation or the accidental operation of nuclear facilities, should be regularly monitored for environmental regulatory compliance. The 135Cs/137Cs isotopic ratios, potentially useful for long-term tracking Cs transport in seawater, can be used as a tool of understanding how radionuclides are transported from different nuclear production source terms and distributed in the ocean. The ultra-high sensitive mass spectrometers (TIMS, SF-ICP-MS and TQ-ICP-MS) have been used to measure the 135Cs/137Cs isotopic ratios. However, the radiochemical separation of Cs from the seawater matrix is essential for the analysis of Cs using the mass spectrometers. An automated radiochemical procedure for the separation of Cs in seawater was developed for the analysis of 135Cs/137Cs isotopic ratios using a sequential column chromatography with AMPPAN and AG50Wx8 cation exchange resins. National Instrument’s LabVIEW is a graphical programming language and a powerful tool for the instrument control. A virtual instrument system for the automated separation of cesium isotopes was developed by the state machine of the fundamental design patterns in LabVIEW. In this study, the conceptual designs of an automated separation system of cesium isotopes, its virtual instrument system based on the LabVIEW state machine architectures and an automated radiochemical procedure were described for the purification of cesium isotopes at trace levels found in seawater discharged from the various nuclear facilities.