• Title/Summary/Keyword: L-Lactate dehydrogenase

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Lactate dehydrogenase activity and isoenzyme distribution in plasma and tissue of Korean native cattle (한우의 혈장 및 조직중의 lactate dehydrogenase의 활성치와 isoenzyme의 분포)

  • Kim, Ki-seog;Cho, Jong-hoo
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.461-467
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    • 1989
  • The activity of lactate dehydrogenase in plasma and various tissues(skeletal muscle, cardiac muscle, liver, lung, kidney and spleen) of Korean native cattle in a Chonju abattoir, the Breeding Stock Farm and Animal Farm of Chonbuk University was determined by using ultra violet method. Using polyacrylamide gel electrophoresis, the lactate dehydrogenase isoenzyme distrimution of plasma and various tissues in Korean native cattle was studied. The plasma lactate dehydrogenase activity of Korean native cattle was $554.80{\pm}92.70IU/l$ and the lactate dehydrogenase activity of male plasma was $543.96{\pm}97.89IU/l$, which was lower than that of female plasma, $579.19{\pm}78.09IU/l$. The plasma lactate dehydrogenase activity of calf was $557.31{\pm}110.27IU/l$ and was no significantly different from that of adult Korean native cattle. But the range of calf lactate dehydrogenase activity was larger than that of adult Korean native cattle. In tissues, the lactate dehydrogenase activity was decreased in order of lung, kidney, spleen, liver, heart and skeletal muscle. The lung had the greatest activity and the skeletal muscle had the least. Lactate dehydrogenase isoenzymes in plasma and tissues were found to have a characteristic distribution and quantitative isoenzyme patterns. In plasma, the LDH1 usually had the greatest activity and other isoenzymes showed a decreasing tendency in order of LDH2, LDH3, LDH4 and LDH5. The distribution of lactate dehydrogenase isoenzymes had a wide variation in tissues. But the distribution of LDH isoenzymes in plasma was similar to that in kidney, and also cardiac muscle and spleen had similar pattern in LDH isoenzymes distribution.

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Electrocatalytic Properties of Metal-dispersed Carbon Paste Electrodes for Reagentless L-lactate Biosensors (금속이 첨가된 탄소전극의 전기화학적 특성과 이를 이용한 L-lactate 바이오센서의 개발)

  • 윤현철;김학성
    • KSBB Journal
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    • v.11 no.4
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    • pp.489-496
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    • 1996
  • Metal dispersed carbon paste electrodes were fabricated, and their electrochemical properties were investigated. Among various metal dispersed carbons, platinum-dispersed carbon paste electrode showed most efficient electrocatalytic characteristics. The overpotential for the oxidation of NADH was significantly lowered in the platinum-dispersed carbon paste electrode, and catalytic current was also enhanced. Based on these electrocatalytic observations, L-lactate biosensor using L-lactate dehydrogenase was constructed to evaluate its performance in terms of sensitivity and stability.

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Purification and Properties of Thermostable L-Lactate Dehydrogenase Produced by Escherichia Coli (대장균으로 부터 생산된 L-lactate Dehydrogenase의 정제 및 특성)

  • Song, Jae-Young;Kim, Kyoug-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.6
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    • pp.964-972
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    • 1994
  • The 4.3-kb gene coding for L-lactate dehydrogenase of Bacillus stearothermophilus has been subcloned and expressed in E. coli cells. The enzyme was purified 200-fold with 25% yield by heat treatment , DEAE-Sephadex, and NAD++ -Sepharose CL-4B affinity chromatography followed by gel filtration through Sephadex G-200 . The molecular weight of the purfied enzyme was estimated to be about 35, 000 and 140, 000 on SDS-polyacrylamide gel electrophoresis and gel filtration, respectively. indicating that the enzyme is composed of four identical subunits. THe enzyme for pyruvate reduction and lactate oxdiation was stable at 60 and 75$^{\circ}C$ for 30 min, and the optimal temperatures for both reactions were 60 and 7$0^{\circ}C$, respectively. The enzyme had an optimal pH at 5.5 and 8.5 in pyruvate reduction and lactate oxidation, respectively. The pH stability of enzyme of pyruvate reduction was table between pH 5 and 7. more than 90% of enzyme activity was lost at 1mM FeSO4 and p-chloromercuribonzoate. The maximal activation of the enzyme was obtained with 0.8mM fructose 1, 6-bisphosphate.

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Cloning and Characterization of the Lactate Dehydrogenase Genes from Lactobacillus sp. RKY2

  • Lee, Jin-Ha;Choi, Mi-Hwa;Park, Ji-Young;Kang, Hee-Kyoung;Ryu, Hwa-Won;Sunwo, Chang-Sin;Wee, Young-Jung;Park, Ki-Deok;Kim, Do-Won;Kim, Do-Man
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.4
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    • pp.318-322
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    • 2004
  • Lactic acid is an environmentally benign organic acid that could be used as a raw material for biodegradable plastics if it can be inexpensively produced by fermentation. Two genes (ldhL and ldhD) encoding the L-(+) and D-(-) lactate dehydrogenases (L-LDH and D-LDH) were cloned from Lactobacillus sp., RKY2, which is a lactic acid hyper-producing bacterium isolated from Kimchi. Open reading frames of ldhL for and ldhD for the L and D-LDH genes were 962 and 998 bp, respectively. Both the L(+)- and D(-)-LDH proteins showed the highest degree of homology with the L- and D-lactate dehydrogenase genes of Lactobacillus plantarum. The conserved residues in the catalytic activity and substrate binding of both LDHs were identified in both enzymes.

Diagnostic Efficiency of Lactate Dehydrogenase, Creatine Kinase and Troponin T in Acute Myocardial Infarction (심근 손상에 있어서 Lactate Dehydrogenase, Creatine Kimase 및 Troponin T 진단적 유용성 비교)

  • Lee, Chae-Hoon;Kim, Kyung-Dong;Kim, Chung-Sook
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.48-55
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    • 1995
  • The present study was designed to evaluate the efficiency of total lactate dehydrogenase, total creatine kinase, LD1/LD2 ratio, CK-MB and newly developed troponin T in acute myocardial infarction. The level of troponin T was $0.01{\pm}0.02{\mu}g/L$ in 34 healthy person, but the peak vaule of acute myocardial infarction ranged in 4.7-24.2 ${\mu}g/L$. Total lactate dehydrogenase was peaked in 1 to 3 days after chest pain and then progressively decreased, but LD1/LD2 ratio was persistently higher than 1.0 for 10 days in most patients. Total creatine kinase and CK-MB were peaked in 1-2 days, and normalized in 3-4 days, so they were useful in early diagnosis of acute myocardial infarction, but not for the late stages of acute myocardial infarction. Troponin T is early elevated and persistently high level for more than 10 days. Comparing with total lactate dehydrogenase, total creatine kinase, LD1/LD2 ratio and CK-MB, troponin-T test improves the efficiency of serodiagnostic method for the detection of ischemic myocardial damage.

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Enzymatic Characterization of Bacillus cereus Lactate Dehydrogenase Isozymes Expressed in Escherichia coli (Bacillus cereus에서 유래한 Lactate Dehydrogenase 동질효소 유전자의 대장균 내 발현 및 효소특성 규명)

  • Jang, Myoung-Uoon;Park, Jung-Mi;Lee, Hong-Gyun;Lee, So-Ra;Kim, Tae-Jip
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.213-218
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    • 2010
  • Lactate dehydrogenases (LDHs) have been highly focused for long time, due to their important roles in biochemical and metabolic pathways of cells. On the basis of genome-wide searching results, three putative LDH genes from Bacillus cereus ATCC 14579 genome have been PCR-amplified, cloned, and well-expressed in E. coli. All three BcLDH isozymes are supposed to share highly conserved catalytic amino acid residues in common $NAD^+$-dependent LDHs. Meanwhile, BcLDH1 consisting of 314 amino acids shares 86 and 49% of identities with BcLDH2 and 3, respectively. Interestingly, only BcLDH1 showed the converting activities between L-lactate and pyruvate in the presence of $NAD^+$ coenzyme, while the other isozymes are likely to have almost no activity. As a result, it was revealed that BcLDH1 can be a typical $NAD^+$-dependent L-lactate-specific dehydrogenase.

Comparison of TNF-Mediated Glucose Catabolism between the TNF-Sensitive and -Resistant Cell Lines

  • Kim, Yeon-Hyang;Park, Bok-Ryun;Cheong, Hee-Sun;Kwon, Oh-Hwan;Kim, Dae-Que;Kim, Soung-Soo
    • BMB Reports
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    • v.32 no.2
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    • pp.140-146
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    • 1999
  • When murine fibrosarcoma L929 cells, a TNF-sensitive cell line, were treated with recombinant human tumor necrosis factor-$\alpha$ (rhTNF-$\alpha$), the activities of glycolytic regulatory enzymes and lactate dehydrogenase increased up to 100-150% compared to the control L929 cells after TNF treatment. By using various metabolic inhibitors and activators, it was found that cAMP-dependent protein kinase is responsible for the increase of activities of the glycolytic enzymes. The activities of glycolytic regulatory enzymes and lactate dehydrogenase of TNF-resistant A549 cells, a human lung carcinoma cell line, did not increase significantly compared to TNF-sensitive L929 cells upon TNF treatment. In contrast, the pyruvate carboxylase activities of A549 cells, but not L929 cells, increased up to 30~40% after TNF treatment. The data suggest that pyruvate carboxylase activity may contribute to the compensation of energy loss mediated by TNF treatment in TNF-resistant A549 cells.

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Protective Effect of Rhodiola sachalinensis A. Bor on Excessive Exercise Stress (홍경천의 과도한 운동스트레스에 대한 방어효과)

  • Lee, Yeonmi;Hong, Hee-Do;Choi, Sang Yoon
    • Journal of Applied Biological Chemistry
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    • v.56 no.1
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    • pp.1-3
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    • 2013
  • Rhodiola sachalinensis A. Bor is a plant naturally grows in a high mountain areas. Aim of this study was to evaluate the effects of R.sachalinensis A. Bor extract against excessive exercise induced stress. R.sachalinensis A. Bor extract significantly inhibited L6 muscle cell death and ATP reduction caused by $H_2O_2$ damage. In addition, the oral administration of R. sachalinensis A. Bor extract in mice improved the 43.9% of treadmill running time and blood profiles of glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactate dehydrogenase, creatinine, lactate when compared to distilled water. These results suggested that R. sachalinensis A. Bor can attenuate excessive exercise induced damage.

Biosynthesis of Chondroitin in Engineered Corynebacterium glutamicum

  • Cheng, Fangyu;Luozhong, Sijin;Yu, Huimin;Guo, Zhigang
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.392-400
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    • 2019
  • Chondroitin, the precursor of chondroitin sulfate, which is an important polysaccharide, has drawn significant attention due to its applications in many fields. In the present study, a heterologous biosynthesis pathway of chondroitin was designed in a GRAS (generally recognized as safe) strain C. glutamicum. CgkfoC and CgkfoA genes with host codon preference were synthesized and driven by promoter Ptac, which was confirmed as a strong promoter via GFPuv reporter assessment. In a lactate dehydrogenase (ldh) deficient host, intracellular chondroitin titer increased from 0.25 to 0.88 g/l compared with that in a wild-type host. Moreover, precursor enhancement via overexpressing precursor synthesizing gene ugdA further improved chondroitin titers to 1.09 g/l. Chondroitin production reached 1.91 g/l with the engineered strain C. glutamicum ${\Delta}L-CgCAU$ in a 5-L fed-batch fermentation with a single distribution $M_w$ of 186 kDa. This work provides an alternative, safe and novel means of producing chondroitin for industrial applications.