• 미생물학회지
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• 제45권1호
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• pp.32-40
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• 2009

수변 낙엽퇴적층에서 호열성 세균의 분포와 물질분해 양상을 파악하기 위하여, 호열성 세균을 분리하고 그 특성을 조사하였다. 고분자 유기물의 분해력이 높고 $60^{\circ}C$에서 생장하는 균주들을 탐색한 결과, 10개의 균주가 선별되었다. 16S rRNA 유전자 염기서열을 결정하고 계통분석한 결과, 모두 Geobacillus 속으로 동정되었다. 균주들의 최적 성장온도는 $55^{\circ}C$와 $60^{\circ}C$, 최적 pH는 초기 pH가 6.0${\sim}$8.0일 때로 나타났으며, 염분 3%일 때를 기준으로 성장속도의 저해가 뚜렷하게 나타났다. 균주의 효소 활성도와 API 50 CHB kit에 의한 탄수화물 이용도를 바탕으로 균주의 생태학적 기능을 알아본 결과, G. stearothermophilus 균주들과 G. kaustophilus 균주들의 특성은 다른 종의 균주들과 명확히 구분되었으며, 나머지 균주들은 최적 성장환경의 생태학적 특징이 다양하게 나타나, 종별 구분과 정확히 일치하지는 않았다. 약산성 환경에서 최적 성장하는 G. stearothermophilus 균주들은 높은 단백질분해효소활성을 보이면서 다당류 분해와 단당류 이용도가 낮았고, 알칼리환경을 최적 서식지로 하는 G. kaustophilus 등은 이와 대조적으로 지질 분해 및 당류의 이용도가 높게 나타났다. 따라서, 수변 낙엽퇴적층에는 호열성 내성포자 형성 세균인 Geobacillus 속 균주들이 다양한 생태학적 지위를 가지며 퇴비화와 유사한 천이과정을 거치면서 수변 퇴적물질을 분해하는데 기여하는 것으로 보여진다.

• Journal of Plant Biotechnology
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• 제40권3호
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• pp.125-134
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• 2013

최근 연구에서 엽록체의 미지의 소기관으로 알려졌던 플라스토글로뷸이 지질 대사에서 중요한 역할을 함이 제시되고 있다. 애기장대 플라스토글로뷸의 프로테옴 단백질 분석은 플라스토글로뷸이 단순히 지질 저장 기관으로써의 기능 뿐아니라 지질 합성 대사에 능동적으로 관여하는 소기관임을 제시하고 있다. 애기장대 플라스토글로뷸에서 34개의 단백질이 발견되었다. 이들을 세 그룹으로 나누어 보면 구조단백질인 플라스토글로불린과 엽록체 대사에 관여하는 효소, 그리고 기능이 미확인된 단백질로 구분 된다. 이들 단백질 유전자의 돌연변이체와 리피도믹스 분석으로 이들 단백질의 기능 규명 연구가 필요하다. 토코페롤 합성의 마지막 단계에 관여하는 VTE1과 VTE4는 엽록체에서 각기 다른 위치에 존재하여 VTE1은 플라스토글로뷸에 존재하나 VTE4는 엽록체 내막에 존재한다. 이 같은 사실은 프레닐퀴논 대사물질이 엽록체 내에서 이동할 가능성을 시사한다. 플라스토글로뷸이 엽록체의 기능을 유지하는데 있어 필수적인지에 대한 유전학 연구가 앞으로 진행되어야 한다. 다양한 스트레스와 발달단계에 따른 플라스토글로뷸의 프로테옴 분석은 지질대사에서 중요한 기능을 하는 신규 단백질을 발견하는데 도움을 줄 것이다. 지금까지 결과로는 플라스토글로뷸은 프레릴퀴논 대사에 있어서 교차로 역할을 함을 제안하고 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6575-6580
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• 2014

This study aimed to evaluate the antimutagenic and anticarcinogenic activity of turmeric essential oil as well as to establish biochemical mechanisms of action. Antimutagenicity testing was accomplished using strains and known mutagens with and without microsomal activation. Anticarcinogenic activity was assessed by topical application of 7, 12 - dimethylbenz[a]anthracene (DMBA) as initiator and 1% croton oil as promoter for the induction of skin papillomas in mice. Inhibition of p450 enzymes by TEO was studied using various resorufins and aminopyrene as substrate. Turmeric essential oil (TEO) showed significant antimutagenic activity (p<0.001) against direct acting mutagens such as sodium azide ($NaN_3$), 4-nitro-O-phenylenediamine (NPD) and N-methyl-N-nitro N'nitrosoguanine (MNNG). TEO was found to have significant antimutagenic effect (>90%) against mutagen needing metabolic activation such as 2-acetamidoflourene (2-AAF). The study also revealed that TEO significantly inhibited (p<0.001) the mutagenicity induced by tobacco extract to Salmonella TA 102 strain. DMBA and croton oil induced papilloma development in mice was found to be delayed and prevented significantly by TEO application. Moreover TEO significantly (P<0.001) inhibited isoforms of cytochrome p450 (CYP1A1, CYP1A2, CYP2B1/2, CYP2A, CYP2B and CYP3A) enzymes in vitro, which are involved in the activation of carcinogens. Results indicated that TEO is antimutagenic and anticarcinogenic and inhibition of enzymes (p450) involved in the activation of carcinogen is one of its mechanisms of action.

• Journal of Microbiology and Biotechnology
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• 제27권10호
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• pp.1867-1876
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• 2017

Most of the biosynthetic pathways for secondary metabolites are influenced by carbon metabolism and supply of cytosolic NADPH. We engineered carbon distribution to the pentose phosphate pathway (PPP) and redesigned the host to produce high levels of NADPH and primary intermediates from the PPP. The main enzymes producing NADPH in the PPP, glucose 6-phosphate dehydrogenase (encoded by zwf1 and zwf2) and 6-phosphogluconate dehydrogenase (encoded by zwf3), were overexpressed with opc encoding a positive allosteric effector essential for Zwf activity in various combinations in Streptomyces lividans TK24. Most S. lividans transformants showed better cell growth and higher concentration of cytosolic NADPH than those of the control, and S. lividans TK24/pWHM3-Z23O2 containing zwf2+zwf3+opc2 showed the highest NADPH concentration but poor sporulation in R2YE medium. S. lividans TK24/pWHM3-Z23O2 in minimal medium showed the maximum growth (6.2 mg/ml) at day 4. Thereafter, a gradual decrease of biomass and a sharp increase of cytosolic NADPH and sedoheptulose 7-phosphate between days 2 and 4 and between days 1 and 3, respectively, were observed. Moreover, S. lividans TK24/pWHM3-Z23O2 produced 0.9 times less actinorhodin but 1.8 times more undecylprodigiosin than the control. These results suggested that the increased NADPH concentration and various intermediates from the PPP specifically triggered undecylprodigiosin biosynthesis that required many precursors and NADPH-dependent reduction reaction. This study is the first report on bespoke metabolic engineering of PPP routes especially suitable for producing secondary metabolites that need diverse primary precursors and NADPH, which is useful information for metabolic engineering in Streptomyces.

• 한국미생물·생명공학회지
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• 제15권2호
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• pp.104-111
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• 1987

Lysine 생산균주를 개량하기 위한 시도로서 Brevibacterium flavum ATCC 21528R 과 Brevibacterium flvum ATCC 21529S의 동종간 및 Brevibacterium flavum ATCC 21528R 과 Corynebacterium glutamicum ATCC 13058S 와 의 이속간 원형질체 융합을 실시하였다. 이들 균주들에 대한 원형질체 형성의 최적조건을 조사하고 재생과 융합에서의 plasma expander의 효과를 검토하였다. 융합주 No. CH23과 No. CH41은 최적 배양조건 하에서 L-lysine 생산성이 모균에서보다 각각 21％와 8.9％ 향상된 것이었다. L-Lysine 생합성 회로상의 중심효소인 asparto-kinase를 포함한 주요효소의 활성을 측정하였고, 융합주 No. CH23과 No. CH41에서의 L-lysine 생합성 대사제어를 친주와 비교하였다.

• 혜화의학회지
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• 제8권1호
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• pp.689-710
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• 1999

Bobitang(BBT) is one of the most important prescription that has been used in oriental medicine(dongyibogam) for recovering spleen condition. The study was done to evaluate effects of BBT water extract on the spleen lipid peroxide content and metabolic enzyme system changes. After pretreatment of BBT I (100mg/kg), BBT II(250mg/kg), BBT III(350mg/kg), BBT IV(500mg/kg) for 1 week, lipid peroxide content and metabolic enzyme system changes of the spleen was measured in 8 months rats. The results were obtained as follows : 1. The content of spleen lipid peroxide was significantly decreased in all experimental groups as compared with control, and best in BBT III IV treated groups. 2. The activity of spleen superoxide generation was significantly decreased in all experimental groups as compared with control, and best in BBT IV III treated groups. 3. The activity of cytochrome P-450 and aminopyrine demethylase wasn't significant change. 4. The activity of aniline hydroxylase was significantly decreased in BBT IV II treated groups, xanthine oxidase was significantly decreased in all experimental groups, aldehyde oxidase was significantly decreased in BBT IV treated group as compared with control. 5. The activity of antioxidant enzymes as superoxide dismutase, catalase, glutathione peroxidase was significantly increased in all experimental groups as compared with control. 6. The activity of glutathion S-transferase was significantly increased in all experimental groups, the concentration of spleen glutathione was significantly increased in BBT IV treated group as compared with control. 7. The activity of ${\gamma}$ -glutamylcystein synthetase was significantly increased in BBT III IV I treated groups as compared with control, the activity of glutathione reductase wasn't significant change. From the above results, BBT is cosidered to have effect of remove peroxide content and free radical that was made during ageing process. It is expected that treatment of BBT can be applied in future clinical study of delaying the ageing process.

• 혜화의학회지
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• 제8권1호
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• pp.625-641
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• 1999

Aging occurs as a part of maturation as the time progresses which manifests in the human body causing morphological and functional degeneration, eventually leading to death. This experimental study was conducted to investigate a herbal formula to fortify the heart with easy clinical applications. Sungshimsan was chosen to study its effects in heart lipid peroxide and metabolic enzyme system in senescence induced rats. After pre-treatment of Sungshimsan for 2 weeks at the dosage of A (100mg/kg), B (250mg/kg), C (350mg/kg), and D (500mg/kg), a lipid peroxide and metabolic enzyme system changes of the heart were meaured in 32 weeks old rats. The following results were obtained in this study: 1. The contents of lipid peroxide was significantly reduced in the experimental groups treated with greater than 2 weeks at 250mg/kg. 2. The enzymatic activity of cytochrome P-450, cytochrome b5, and NADPH-cytochrome P450 reductase were significantly decreased in the 250mg/kg, 350mg/kg, and 500mg/kg experimental groups. 3. The activity of glutathione and glutathione S-transferase were significantly increased in the 250mg/kg, 350mg/kg, and 500mg/kg experimental groups. 4. The activity of glutathione reductase and glutathione peroxidase were not influenced compared to the control group. 5. The activity of ${\gamma}$-glutamylcystein synthetase was significantly increased in the 250mg/kg, 350mg/kg, and 500mg/kg experimental groups. 6. The activity of enzymes detoxificatioon superoxide dismutase and catalase were not influenced compared to the control group. Summarizing above results suggest that the Sungshimsan has profound effects in the heart lipid peroxide, free radicals, and delaying the heart aging process. Further clinical researches and application can be anticipated on the topic of senility and gerontology.

• KSBB Journal
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• 제22권5호
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• pp.345-350
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• 2007

인산당은 모든 유기체에서 발견되며 무척 다양한 유용성을 지니고 있다. 특히 glucose 1-phosphate (G1P), glucose 6-phosphate (G6P), fructose 6-phosphate (F6P) 등은 해당과 정, 당합성과정, 5탄당 인산화과정 및 캘빈회로와 같은 탄수화물 대사와 에너지 생산 대사의 주요한 핵심 중간물질이다. 특히 해당과정에서 F6P는 G6P의 이성질화반응에 의하여 생성된다. F6P의 대량생산은 전분을 이용하는 것이 가능한데, 우선 전분에 인산화효소를 가하여 G1P를 얻고, 이 G1P를 자리옮김효소 (phosphoglucomutase, GM)와 이성질화효소 (phosphoglucoisomerase, GI)를 순차적으로 적용하여 G6P와 F6P를 생산하게 된다. 효소반응의 경우 전분의 용해도 증가, 반응속도의 향상 및 미생물의 오염방지 등을 위하여 중온성 효소보다는 고온성 효소 혹은 내열성 효소가 선호된다. 본 연구는 세 가지 내열성 효소를 이용하여 전분으로부터 두 단계반응으로 F6P를 생산하는 것에 관한 것이다. 실험에 사용된 효소는 대장균에서 발현된 재조합 효소로서, 효소의 생산은 유가식 배양을 이용하였다. 1.2% 가용성 전분 200 L를 이용하여 1,253 g의 순수한 G1P를 생산하였으며 이를 이용하여 최종적으로 30% 수율로 F6P를 생산할 수 있었다. 최대수율을 얻기 위하여 반응표면분석법을 이용하여 GM : GI = 1 : 1.23, 63.5$^{\circ}C$, pH 6.85의 조건이 도출되었으며, 이 조건하에서 실험을 통하여 20 g/L의 전분을 이용하여 30% 수율로 F6P가 생성됨을 확인할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권7호
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• pp.1003-1010
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• 2008

The objective of the present study was to identify differences in the expression levels of liver proteins between healthy and ketotic cows, establish a liver metabolic interrelationship of ketosis and elucidate the metabolic characteristics of the liver during ketosis. Liver samples from 8 healthy multiparous Hostein cows and 8 ketotic cows were pooled by health status and the proteins were separated by two-dimensional-electrophoresis (2D-E). Statistical analysis of gels was performed using PDQuest software 8.0. The differences in the expression levels of liver proteins (p<0.05) between ketotic and healthy cows were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry. Five enzymes/proteins were identified as being differentially expressed in the livers of ketotic cows: expression of 3-hydroxyacyl-CoA dehydrogenase type-2 (HCDH), acetyl-coenzyme A acetyltransferase 2 (ACAT) and elongation factor Tu (EF-Tu) were down-regulated, whereas that of alpha-enolase and creatine kinase were up-regulated. On the basis of this evidence, it could be presumed that the decreased expression of HCDH, which is caused by high concentrations of acetyl-CoA in hepatic cells, in the livers of ketotic cows, implies reduced fatty acid ??oxidation. The resultant high concentrations of acetyl-CoA and acetoacetyl CoA would depress the level of ACAT and generate more ??hydroxybutyric acid; high concentrations of acetyl-CoA would also accelerate the Krebs Cycle and produce more ATP, which is stored as phosphocreatine, as a consequence of increased expression of creatine kinase. The low expression level of elongation factor Tu in the livers of ketotic cows indicates decreased levels of protein synthesis due to the limited availability of amino acids, because the most glucogenic amino acids sustain the glyconeogenesis pathway; thus increasing the level of alpha-enolase. Decreased protein synthesis also promotes the conversion of amino acids to oxaloacetate, which drives the Krebs Cycle under conditions of high levels of acetyl-CoA. It is concluded that the livers of ketotic cows possess high concentrations of acetyl-CoA, which through negative feedback inhibited fatty acid oxidation; show decreased fatty acid oxidation, ketogenesis and protein synthesis; and increased gluconeogenesis and energy production.

• Journal of Ginseng Research
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• 제41권1호
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• pp.60-68
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• 2017

Background: Various Panax ginseng cultivars exhibit a range of diversity for morphological and physiological traits. However, there are few studies on diversity of metabolic profiles and genetic background to understand the complex metabolic pathway in ginseng. Methods: To understand the complex metabolic pathway and related genes in ginseng, we tried to conduct integrated analysis of primary metabolite profiles and related gene expression using five ginseng cultivars showing different morphology. We investigated primary metabolite profiles via gas chromatography-mass spectrometry (GC-MS) and analyzed transcriptomes by Illumina sequencing using adventitious roots grown under the same conditions to elucidate the differences in metabolism underlying such genetic diversity. Results: GC-MS analysis revealed that primary metabolite profiling allowed us to classify the five cultivars into three independent groups and the grouping was also explained by eight major primary metabolites as biomarkers. We selected three cultivars (Chunpoong, Cheongsun, and Sunhyang) to represent each group and analyzed their transcriptomes. We inspected 100 unigenes involved in seven primary metabolite biosynthesis pathways and found that 21 unigenes encoding 15 enzymes were differentially expressed among the three cultivars. Integrated analysis of transcriptomes and metabolomes revealed that the ginseng cultivars differ in primary metabolites as well as in the putative genes involved in the complex process of primary metabolic pathways. Conclusion: Our data derived from this integrated analysis provide insights into the underlying complexity of genes and metabolites that co-regulate flux through these pathways in ginseng.