• 미생물학회지
• /
• 제26권3호
• /
• pp.197-206
• /
• 1988

Klebsiella pneumoniae의 세포외막으로부터 2-furaldehyde를 2-furoic acid로 산화시키는 2-furaldehyde dehydrogenase를 분리하여 그 특성을 조사하였다. 이 효소는 $\beta$-$NAD^{+}$를 특이적으로 요구하였다. 분리과정중의 효소활성도는 2-furaldehyde를 기질로 사용하고 $\beta$-$NAD^{+}$를 조효소로 사용하면서 high performance liquid chromatography에 의해 측정 되었다. 세포외막은 Percoll의 밀도흉배에 의한 초원심분리방법과 $Mg^{2+}$, Triton X-100으로 용해시킨 후, 초원심분리시키는 방법으로 수집되었다. 세포외막단백질은 EDTA와 lysozyme을 처리함으로서 얻어졌고, 효소는 QAE-Sephadex Q-504 S Sephadex G-100-을 사용하면서 column chromatography 방법에 의해 분리되었다. 본 효소는 $85^{\circ}C$, PH9.5, 그리고 1.5% (vol/vol) Triton X-100의 존재하에서 최대활성을 보여주었다. 효소의 분자량은 nondenaturing polyacrylamide gel e electrophoresis의 결과, 88, 000.으로 추정되었고, 2-furaldehyde에 대한 효소의 Km값은 4.72 mM 이였다.

• 한국수산과학회지
• /
• 제27권1호
• /
• pp.83-88
• /
• 1994

연어, 산천어, 무지개송어 등 우리나라의 연어과 어류에 있어서 종의 식별 및 3배체 어류의 판정에 동위효소를 genetic marker로 활용할 수 있는지의 가능성을 타진하고자 LDH, MDH, IDH, a-GPDH, ME, 6-PGD, PGI 및 PGM 등 8개 동위효소에 대하여 골격근 조직을 중심으로 분석을 실시하였다. 이중 골격근 조직의 MDH-B와 IDH loci에서 종간에 뚜렷한 차이를 나타내었으며 특히 MDH-B loci의 b 유전자의 출현빈도는 연어나 산천어에서는 거의 나타나지 않았으나 무지개송어에서는 매우 높게 나타났다. 또한 IDH도 무지개송어와 산천어간의 genetic marker로 유용할 것으로 보인다. 한편, PGI는 3배체 어류 생산시 친어를 상호 대립유전자(allele)의 동형접합(homozygote)인 개체를 사용할 경우 이의 효율적인 판정을 위한 새로운 marker로 활용할 수 있는 가능성을 가지고 있는 것으로 나타났으며 다른 loci에서는 별다른 차이를 발견할 수가 없었다.

• BMB Reports
• /
• 제29권6호
• /
• pp.493-499
• /
• 1996

The biochemical properties of purified D-hydruxyisovalerate dehydrogenase from Fusarium sambucinum was elucidated. D-Hydroxyisovalerate dehydrogenase produced solely D-hydroxyisovalerate from 2-ketoisovalerate. The isoelectric point of the purified enzyme was 7.0. The enzyme was highly specific with 2-ketoisovalerate ($K_{m}=0.188$ mM, $V_{max}=8.814$ mmol/min mg) and 2-keto-3-methyl-n-valerate ($K_{m}=0.4$ mM, $V_{max}=1.851$ mmol/min mg) for the reductive reaction. This was also seen by comparing D-hydroxyisovalerate ($K_{m}=1.667$ mM, $V_{max}=0.407$ mmol/min mg) and D-hydroxy-3-methyl-n-valerate ($K_{m}=6.7$ mM, $V_{max}=0.648$ mmol/min mg) for the oxidative reaction. Thiol blocking reagents, such as iodoacetamide, N-ethylmaleimide and p-chloromecuribenzoate inhibited about 80% of enzyme activity at 0.02 mM, 50 mM and 50 mM, respectively. The enzyme activity was also inhibited by the addition of 0.1 mM of various metal ions, such as $Fe^{2+}$ (67%), $Cu^{2+}$ (88%), $Zn^{2+}$ t (76%) and $Mg^{2+}$ (9%). The enzyme was stable over three months in 50 mM potassium phosphate buffer (pH 5~7) at $-80^{\circ}C$. However the purified enzyme lost 30% of its activity in the same buffer after 24 h at $4^{\circ}C$. The studies about thermal inactivation of D-hydroxyisovalerate dehydrogenase exhibit 209.2 kJ/M of activation enthalpy and 0.35 kJ/mol K of activation entropy.

• Journal of Ginseng Research
• /
• 제22권1호
• /
• pp.51-59
• /
• 1998

In this study, rat hepatocytes known to have active glucose metabolism were obtained to investigate the hypoglycemic action of fat soluble fraction of red ginseng by using the liver perfusion technique and incubated in two different media-one containing insulin and glucagon (control group), and the other containing glucagon only The activities of main regulating enzymes, such as glucokinase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenate, and glucose 6-phosphatase, related to metabolic pathways of glucose in these two kinds of hepatocytes were compared between these two groups and the effects of addition of fat soluble fraction ($10^1$~$10^4$%) from red ginseng to these two groups on these enzymes were also detected. The results were as follows. The specific activity of enzymes such as glucokinase, flucorse 6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase related to glucose-consuming pathways of insulin-deficient group was much less than control one. However, their decreased activity was recovered after the addition of fat-soluble fraction at all range of concentrations. The specific activity of these enzymes after the addition of ginseng components to the control group was also increased. On the other hand, the specific activity of glucose 6-phosphatase related to glucose-producing pathway of insulin-deficient group was much higher than control one, but their increased activity was decreased obviously after the addition of fat soluble fraction at all range of concentrations. The same results were observed after the addition of fat-soluble fraction to the control group. These results suggest that the red ginseng saponin components might be effective on diabetic hyperglycemia by regulating the activity of enzymes related to glucose metabolism directly and/or indirectly. The effects of fat-soluble fraction ($10^2$%) and ginsenosides (mixture, $Rb_1$ and $Rg_1$, $10^4$%) on hypoglycemic action were compared. As a result, they showed considerable effect on hyperglycemia, but the best eff ect on the activities of glucokinase and glucose 6-phosphate dehydrogenase was appeared by ginsenoside $Rb_1$ and that of 6-phosphogluconate dehydrogenase and glucose 6-phosphatase was by ginsenoside mixture.

• Applied Biological Chemistry
• /
• 제40권5호
• /
• pp.447-450
• /
• 1997

18 종의 N-치환 phenyl 1,3,5-trimethylpyrazole-4-carboxamide 를 합성하여 벼 잎집무늬 마름병균 Rhizoctonia solani $K{\ddot{u}}hn$ 에 대한 균사생육억제력 $(pEC_{50})$ 과 이 균에서 분리한 succinate dehydrogenase (SDH)의 효소활성 저해력 $(pI_{50})$ 을 측정하였다. 구조-활성 상관연구를 통해 균사생육억제력이 SDH 효소저해력과 소수성인자인 chromatographic capacity factor $(\acute{k})$ 에 의해 79%까지 유의성 있게 해석되었으며 이는 합성한 화합물의 주작용점이 SDH 이고 생체내에서 작용점까지 도달하는데 필요한 소수성성질이 살균력의 발현에 중요한 인자임을 보여준다.

• Journal of Microbiology
• /
• 제38권4호
• /
• pp.275-280
• /
• 2000

Pseudomonas sp. S-47 is capable of catabolizing 4-chlorobenzoate (4CBA) as rarbon and energy sources under aerobic conditions via the mesa-cleavage pathway. 4CBA-dioxygenase and 4CBA-dihydrodiol dehydrogenase (4CBA-DD) catalyzed the degradation af 4CBA to produce 4-chlorocatechol in the pathway. In this study, the xylL gene encoding 4CBA-DD was cloned from the chromosomal DNA of Pseudomonas sp. S-47 and its nucleotide sequence was analyzed. The xylL gene was found to be composed of 777 nucleotide pairs and to encode a polypeptide of 28 kDa with 258 amino acid residues. The deduced amino acid sequence of the dehydrogenase (XylL) from strain S-47 exhibited 98% and 60% homologies with these of the corresponding enzymes, Pseudomonas putida mt-2 (XyIL) and Acinetobacter calcoaceticus (BenD), respectively. However, the amino arid sequences show 30% or less homology with those of Pseudomonas putida (BnzE), Pseudomonas putida Fl (TodD), Pseudomonas pseudoalcaligenes KF707 (BphB), and Pseudomonas sp. C18 (NahB). Therefore, the 4CBA-dihydrodiol dehdrogenase of strain S-47 belongs to the group I dehydrogenase involved in the degradation of mono-aryls with a carboxyl group.

• Current Research on Agriculture and Life Sciences
• /
• 제5권
• /
• pp.68-74
• /
• 1987

A. calcoaceticus C10은 CL 배지에서 ${\varepsilon}$-caprolactone 및 succinate에 의해 생육에 아무 영향도 받지 않았으나 adipate와 xylose에 의해서 균의 생육도가 증가하였다. 특히 이 균이 탄소원으로 이용할 수 없는 포도당에 의해서도 생육도가 증가하였으며 6시간 배양 후 0.2%의 포도당을 첨가한 경우에는 16시간 후 CL 배지에서 생육한 것보다 약 2배의 생육도를 냐타내었다. A. calcoacelicus C10이 생산하는 cyclohexanol dehydrogenase(CDH)는 ${\varepsilon}$-caprolactone, succinate, xylose 및 포도당에 의해 이화물 억제를 받지 않았으나 사이클로헥사놀 대사경로의 최종생산물인 adipate에 의하여 생합성 억제를 받았다. CL 배지에 0.1%의 adipate를 첨가한 배지에서는 CDH가 다소 유도되었으나 0.2%의 adipate를 첨가한 배지에서는 CDH가 거의 유도되지 않아 0.5%의 adipate를 첨가한 것과 비슷한 억제효과를 나타내었다.

• Toxicological Research
• /
• 제35권1호
• /
• pp.13-24
• /
• 2019

Numerous ethnomedicinal uses have been attributed to different parts of Annona senegalensis (ASE), including its uses as food and food additives. The present study investigated toxicological and antioxidant effects of 28 days administration of ethanol extracts of ASE stem bark to Wistar strain albino rats. Acute toxicity test was done to determine lethal dose in Wistar rats while sub-acute toxicity test was conducted on rats divided into four groups (A - control, B - 50 mg/kg, C - 100 mg/kg, D - 150 mg/kg, respectively and treated for 28 days. Oxidative stress markers in liver and kidney as well as hepatic succinate dehydrogenase activity in the mitochondrial and post mitochondrial fractions (PMF) were evaluated. The $LD_{50}$ value of ASE was > 2,000 mg/kg. White blood cell counts gradually increased, but red blood cell counts and haematocrits level decreased significantly (p < 0.05) by about 50%. Liver enzymes in the serum and mitochondrial succinate dehydrogenase activity increased significantly (p < 0.05). Superoxide dismutase and catalase activities also increased in liver mitochondria and PMF while malondialdehyde (MDA) and reduced glutathione levels increased only in the PMF. Furthermore, only MDA levels increased significantly in the kidney after 28 days extract administration. Histopathological examination showed hepatic necrosis and no obvious signs of nephrotoxicity. Anona senegalensis is relatively safe, but prolonged ingestion could induce oxidative stress and impair ATP synthesis through the modulation of the activity of mitochondrial succinate dehydrogenase.

• 한국식품영양과학회지
• /
• 제34권10호
• /
• pp.1599-1605
• /
• 2005

본 연구는 냉장우육과 냉동우육의 판별법을 개발하기 위하여, 한우 육을 부위별(사태, 등심, 우둔, 양지 )로 구입하고 냉장(4$\pm$ 1$^{\circ}C$), 냉동(-4, -18, -77$^{\circ}C$)상태로 15일 저장하면서 압착육즙채취기를 이용하여 근육 세포내 mitochondria 막에 존재하는 mitochondrial malate dehydrogenase활성 특성을 비교 연구한 결과 다음과 같은 결과를 얻었다. 본 실험을 위하여 제작된 압착육즙채취기를 이용하여 정량적인 압착 육즙을 얻을 수 있었고, 1.5g의 우육을 이용하였을 때 0.15mL의 압착육즙을 얻었다. 냉장우육과 냉동우육 간의 mite-chondral malate dehyogenase의 각 부위별 활성은 사태의 경우 냉장우육은 23.63 Unit/mL의 활성을 나타내었고,-4$^{\circ}C$는 20.91 Unit/mL, -18$^{\circ}C$는 26.43 Unit/mL 그리고 -77$^{\circ}C$는 25.90 Unit/mL의 활성을 나타내었다. 동일한 부위에서 저장온도에 따른 효소의 활성도는 냉장우육보다 -4$^{\circ}C$를 제외한 법동 저장 온도(-18, -77$^{\circ}C$)에서 모두 유의적으로 높은 효소의 활성을 나타내었다(p < 0.05). 동일한 저장 온도에서 각부위 간에 활성의 차이점은 -77$^{\circ}C$에서 냉동 저장한 우육이 유의적 차이 없이 가장 높은 효소 활성을 나타내었고,-77$^{\circ}C$ 저장온도를 제외하고는 모두 유의성 있는 변화량을 나타내었다(p < 0.05). 실험군의 압착육즙 내 mitochondrial malate dehydrogenase의 활성은 10분간 흡광도 변화량을 조사함으로써 계산되었다. 등심을 제외한 모든 부위에서 호소반응개시 3분 후에 -4$^{\circ}C$에서 동결우육을 제외하고 동결우육의 효소 활성이 떨어지는 것을 확인할 수 있었고, 5분 반응 후에는 그 효소의 활성을 발견할 수 없었다. 냉장우육은 12분까지 활성을 측정할 수 있었다. 15일간 냉장우육 및 냉동우육을 저장 온도에 따라 저장하면서 그 활성을 측정하였는데, 저장 기간이 길어져도 그 효소활성의 차이는 유의적으로 변화하지 않고, 모든 실험군(사태, 등심, 양지, 우둔 부위 )에서 유의적으로 유사한 활성을 유지하였다(p < 0.05). 따라서 본 연구 결과로부터 mitochondrial malate dehydrogenase의 활성을 이용하여 냉장우육과 냉동우육 유무를 판별하는데 유효한 지표로 사용 가능한 것으로 사료되었다.

• Journal of Microbiology and Biotechnology
• /
• 제6권2호
• /
• pp.142-144
• /
• 1996

To clone the gene coding for steroid $\Delta^1$-dehydrogenase of Arthrobacter simplex, its genomic library was constructed with a , $\lambda$gt11 expression vector and immunoscreened with antiserum against the enzyme. One positive clone was found to carry a 1.6-kb EcoR I restriction endonuclease fragment of A. simplex DNA. The restriction map of the 1.6-kb EcoR I fragment was determined after cloning of the DNA into pBS vector.