• Asian-Australasian Journal of Animal Sciences
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• 제14권12호
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• pp.1670-1674
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• 2001

To understand molecular mechanisms that regulate deposition and release of intramuscular fat, a fasting-induced clone was identified by differential screening from cDNA library of adipose tissues of Korean cattle. The clone had a total length of 1,319 nucleotides coding for 334 amino acids. It was identified as one encoding L-lactate dehydrogenase H chain (LDH-B). Comparison of the deduced amino acid sequences of bovine LDH-B with those of pig, human, rat, and mouse showed 98%, 98%, 97%, and 96% identity, respectively. Food deprivation for 48 h increased mRNA levels of LDH-B gene in adipose tissues of Korean cattle compared to fed- and 6 h refed- tissues. The expression of obese mRNA was examined for individual adipose tissue from several fat depots. Fasting induced expression of LDH-B gene in subcutaneous adipose tissues, but it did not affect expression levels in abdominal, perirenal and intramuscular tissues. Results demonstrate that induction of LDH-B gene during fasting may represent a metabolic shift from anaerobic state to aerobic predominance in fasted adipose tissues and that its responses to fasting are different among several adipose tissues.

• 한국동물학회지
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• 제23권4호
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• pp.263-272
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• 1980

개구리 2種 (Rana nigromaculata와 Rana plancyi chosenica)과 도룡뇽 (Hynobius leechii)의 胚發生에 EK른 lactate dehydrogenase (LDH)와 malate dehydrogenase (MDH)의 isozyme 組成變化를 polyacrylamide 電氣泳動法으로 조사 분석하고 이를 成體의 몇 器官과 비교하였다. R. nigromaculata에서 LDH의 B subunit의 合成을 지배하는 유전자는 heterozygous이고 H. leechii에서는 A subunit의 合成을 지배하는 유전자가 heterozygousfk고 추정된다. 위의 3種의 兩棲類\ulcorner 胚에서 發生初期에는 LDH-1 (심장형)의 活性이 높으나 發生이 진행됨에 따라 LDH-5 (근육형)의 活性도 점차 증가된다. MDH의 경우 發生初期부터 MDH-m과 MDH-s가 존재하고 발생 全段階를 통하여 그 組成에는 변화가 없으나 MDH-m의 活性이 점차 증가하는 경향을 보인다.

• 한국식품영양과학회지
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• 제23권6호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제31권4호
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• pp.537-547
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• 2018

Objective: This study was performed to understand transcriptional changes in the genes involved in gluconeogenesis and glycolysis pathways following castration of bulls. Methods: Twenty Korean bulls were weaned at average 3 months of age, and castrated at 6 months. Liver tissues were collected from bulls (n = 10) and steers (n = 10) of Korean cattle, and hepatic gene expression levels were measured using quantitative real-time polymerase chain reaction. We examined hepatic transcription levels of genes encoding enzymes for irreversible reactions in both gluconeogenesis and glycolysis as well as genes encoding enzymes for the utilization of several glucogenic substrates. Correlations between hepatic gene expression and carcass characteristics were performed to understand their associations. Results: Castration increased the mRNA (3.6 fold; p<0.01) and protein levels (1.4 fold; p<0.05) of pyruvate carboxylase and mitochondrial phosphoenolpyruvate carboxykinase genes (1.7 fold; p<0.05). Hepatic mRNA levels of genes encoding the glycolysis enzymes were not changed by castration. Castration increased mRNA levels of both lactate dehydrogenase A (1.5 fold; p<0.05) and lactate dehydrogenase B (2.2 fold; p<0.01) genes for lactate utilization. Castration increased mRNA levels of glycerol kinase (2.7 fold; p<0.05) and glycerol-3-phosphate dehydrogenase 1 (1.5 fold; p<0.05) genes for glycerol utilization. Castration also increased mRNA levels of propionyl-CoA carboxylase beta (mitochondrial) (3.5 fold; p<0.01) and acyl-CoA synthetase short chain family member 3 (1.3 fold; p = 0.06) genes for propionate incorporation. Conclusion: Castration increases transcription levels of critical genes coding for enzymes involved in irreversible gluconeogenesis reactions from pyruvate to glucose and enzymes responsible for incorporation of glucogenic substrates including lactate, glycerol, and propionate. Hepatic gluconeogenic gene expression levels were associated with intramuscular fat deposition.

• 미생물학회지
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• 제46권2호
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• pp.213-218
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• 2010

Lactate dehydrogenases (LDHs)는 세포 내의 생화학적 대사경로에서 중요한 역할을 담당하는 효소로서 오랜 동안 많은 관심을 받았다. 본 연구에서는 다양한 미생물 genome database의 탐색을 통해 Bacillus cereus ATCC 14579 genome으로부터 LDH로 추정되는 3종의 유전자를 발견하고, 대장균에서 클로닝 및 대량 발현하였다. 모든 BcLDH 동질효소들의 상동부위 아미노산 잔기 대부분이 기존의 $NAD^+$-의존형 LDH와 높은 상동성을 보였다. 한편 314개의 아미노산으로 이루어진 BcLDH1과 2는 86%의 서열 상동성을 보였으나, BcLDH3와는 49%의 상동성을 나타냈다. 흥미롭게도 BcLDH1만이 $NAD^+$ 조효소 존재 하에서 L-lactate와 pyruvate 간의 상호전환 활성을 나타냈으며, 그 외의 동질효소들은 거의 활성을 보이지 않았다. 결론적으로 BcLDH1은 전형적인 $NAD^+$-의존형이며, L-lactate에 특이적인 dehydrogenase 효소임을 확인하였다.

• 생명과학회지
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• 제20권3호
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• pp.416-423
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• 2010

열대 저산소 환경에 적응되어 있는 angelfish (Pterophyllum scalare)를 급격한 온도변화($27{\pm}0.5{\rightarrow}18{\pm}0.5^{\circ}C$) 및 DO 변화($6{\pm}1{\rightarrow}18\;ppm$)에 2시간 동안 적응시킨 후 젖산탈수소효소(EC 1.1.1.27, lactate dehydrogenase, LDH) 동위효소의 특성 및 유전자발현을 연구하였다. LDH 동위효소의 특성은 native-polyacrylamide gel 전기영동, Western blot 분석 및 효소활성 측정으로 확인하였다. 전기영동 결과 liver- 및 eye-specific Ldh-C 유전자는 간, 눈 및 뇌 조직에서 발현되었다. Western blot 분석 결과 LDH $A_4$ 동위효소는 $B_4$ 동위효소보다 음극 쪽에 나타났다. 간 조직에서 온도 저하 시 LDH $A_4$ 동위효소가 증가하고 $B_4$ 동위효소는 감소하였으며, DO 증가 시 LDH $A_4$ 및 $C_4$ 동위효소가 증가하고 $B_4$ 동위효소는 감소하였다. 눈 조직에서는 온도 저하 시 LDH $A_4$ 및 $B_4$ 동위효소가 증가하고 $C_4$ 동위효소는 감소하였으며, DO 증가 시 LDH $A_4$ 및 $B_4$ 동위효소는 증가하지만 $C_4$ 동위효소 및 하부단위체 C를 포함하는 동위효소는 감소하였다. 심장 조직에서는 DO 증가 시 LDH 활성이 증가하였고, LDH $B_4$ 동위효소가 증가하였다. 뇌 조직에서는 온도 저하 시 LDH $A_4$ 및 $B_4$ 동위효소가 증가하였고, DO 증가 시 LDH $B_4$ 동위효소는 증가하였다. 따라서 liver- 및 eye-specific Ldh-C는 DO 변화에 의해 영향을 받으며 간 및 눈 조직에서 LDH $B_4$ 및 $C_4$ 동위효소는 서로 상대적으로 조절되므로 $C_4$ 동위효소는 lactate oxidase로서 기능을 나타내는 것으로 사료된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제15권2호
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• pp.137-143
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• 2007

Lactate dehydrogenase (LDH) is a ubiquitous enzyme that plays a significant role in the clinical diagnosis of pathologic processes. Discovery of the LDH (BmLDH) gene in B. mori may shed light on its role in the biology of Lepidoptera species, and afford further understanding of the function of the enzyme. In this study, we used the bioinformatics tools to identify LDH gene in B. mori. Sequence analysis showed that BmLDH cDNA contains a 996 bp open reading frame, encoding 331 AA proteins, with seven introns. Compared with hHLDH (human heart LDH), BmLDH contained the same key active sites. Domain search and protein fold recognition analyses provide compelling evidences that the deduced protein is a LDH. Using the computer program MEGA3, we conducted a search for homologs of BmLDH among many eukaryotic species and confirmed that the BmLDH was conserved in all organisms investigated. This gene has been registered in GenBank under the accession number EU000385.

• 동의생리병리학회지
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• 제21권1호
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• pp.98-103
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• 2007

In the present study, the author intended to investigate whether Gamiyukgunja-tang (Jiaweiliujunzi-tang, GYGT) significantly affect both mucin release from and MUC5AC gene expression in cultured hamster tracheal surface epithelial (HTSE) cells. Confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of GYGT to assess the effect on 3H-mucin release. Possible cytotoxicity of the agent was assessed by measuring lactate dehydrogenase (LDH) release. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analysed and effect of GYGT on MUC5AC gene expression in cultured HTSE cells were investigated. GYGT did not affect mucin release from cultured HTSE cells. GYGT did not show significant cytotoxicity. GYGT also did not affect the secretion of the other releasable glycoproteins with less molecular weight than mucin. GYGT increased the expression level of MUC5AC gene. We suggest that the effect of GYGT with their components should be further investigated through ongoing research.

• 대한한방내과학회지
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• 제28권4호
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• pp.797-807
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• 2007

Objectives : The purpose of this study was to investigate whether Mahwangyunpye-tang(MYT) significantly affects mucin secretion from airway epithelial cells. Methods : Confluent hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of MYT to assess the effect of the agent on 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. Effect of MYT on contractility of isolated tracheal smooth muscle was investigated; also investigated was effect of the agent on MUC5AC gene expression in cultured NCI-H292 cells. Possible cytotoxicities of the agent were assessed both by measuring lactate dehydrogenase (LDH) release from HTSE cells and examining the rate of survival and proliferation of NCI-H292 cells. Results : MYT significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity. MYT chiefly affected the 'mucin' secretion. MYT inhibited Acetylcholine-induced contraction of isolated tracheal smooth muscle. MYT did not significantly affect the expression levels of MUC 5AC gene in cultured NCI-H292 cells. Conclusions : Based on the above results, it is suggested that MYT increased mucin secretion from cultured HTSE cells without significant cytotoxicity and inhibited contraction of isolated tracheal smooth muscle.

• 대한의생명과학회지
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• 제11권3호
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• pp.319-326
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• 2005

It is well known that oxidative stress of reactive oxygen species (ROS) may be a causative factor in the pathenogenesis of bone disorder on osteoblast or osteoclast. The purpose of this study was to evaluate the cytotoxicity of oxidative stress, protective effect of glutamate receptor antagoinst against ROS-induced osteotoxicity, secretion of tumor necrosis factor $(TNF)-\alpha$ and the expression of c-fos gene in the cultured rat osteoblasts and osteoclasts. Cell viability by MTS assay or !NT assay, activity of glutathione peroxidase (GPx), lipid peroxidation (LPO) activity, protein synthesis by sulforhodamine B (SRB) assay, alkaline phosphatase (ALP) activity, lactate dehydrogenase (LDH) activity, MTS assay for NMDA (N-methyl-D-aspartate) receptor antagonist or AMPA/kainate receptor antagonist, measurement for $TNF-\alpha$, and c-fos gene expression were performed after these cells were treated with or without various cocentrations of xanthine oxidase (XO), hypoxanthine (HX), D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), respectively. In this study, XO/HX showed decreased cell viability and glutathione peroxidase (GPx) activity, but it showed increased LPO activity, $TNF-\alpha$ secretion and c-fos expression. APV and CKA incresed protein sythesis and ALP activity. While, CNQX or DNQX did not show any protective effect in LDH activity or cell viability. From these results, XO/HX showed cytotoxic effect in cultured rat osteoblast or osteoclast, and also NMDA receptor antagonist such as APV or CKA was effective in blocking XO/HX-induced osteotoxicity in these cultures.