• 한국응용곤충학회지
• /
• 제34권3호
• /
• pp.181-190
• /
• 1995

암컷 Aedes aegypti의 난성숙과장에서 새로 나타나는 malate dehydrogenase(L-malate, $NAD^+$ oxidoreductase, EC 1.11.37, MDH)를 DEAE-Sepharose, Sulphonyl-Sepharose, Cibacron 3FGA affinity chromatography를 이용하여 분리정제하여 그 특성을 조사하였다. 분자량은 70,000 dalton정도의 dimer 형태로 되어 있으며 최적 pH는 malate-oxaloacetate반응에서는 pH 9.0~9.2, oxaloacetate-malate 반응에서는 pH 9.8~10.2이었다. 정재된 MDH는 mitochondria에 위치하고 있으며 기질로서 malate에 대한 Km값의 경우 $1.29 \times 10^{-4}$ M, oxaloacetate에 대한 Km 값은 $6.58\times 10^{-4}$M, NAD에 대한 Km값은 $0.76\times 10^{-3}$ M이며 NADH에 대한 Km 값은 $3.8\times 10^{-3}$ M 을 보이고 있으며 각각의 기질에 의한 저해현상을 보이고 있었다. 기질에 대한 Km값을 부분적으로 분리한 DEAE-sepharose에 흡착된 원형질 MDH와 비교한 결과 malate에 대한 Km 이 $8.92\times 10^{-3}$으로 상당한 차이를 보이고 있었다. 또한 정제된 MDH는 cltrate, $\alpha$-ketoglutarate, ATP 등의 대사산물에 의하여 저해작용을 받았다. ATP 및 citrate에 의한 MDH 활성도 저해는 oxaloacetate-malate반응에서 보다는 malate-oxaloacetate 반응에서 덜 일어났다. Oxaloacetate-malate 반응의 경우 ATP에 의하여저해작용이 완전히 일어났으며 malate-oxaloacetate반응에서는 cltrate에 의하여 저해작용이 일어나지 않았다. 흡혈 후 생성되는 MDH는 난소에서 합성되며 흡혈 수 난소에서 18시간 때부터 활성도가 나타나 48시간 이후 최고 활성도가 유지되는데 TCA회로의 isocitrate dehydrogenase 의 경우 난소내에서의 활성도 변화가 MDH의 변화 양상과 같았다.

• KSBB Journal
• /
• 제9권3호
• /
• pp.319-324
• /
• 1994

옥수수잎의 유관속초세포 내에 존재하는 malate dehydrogenase(EC 1.1.1.40)을 이용하여 malate 정량용 biosensor를 제작한 후, 기질인 malate에 대하여 여러 가지 조건하에서 측정한 결과, sodium-alginate로 고정화한 것이 하지 않은 것보다 180시 간까지 지나도 40% 이상의 activity를 가지며, 최적 pH는 8.0이었으며 pH 7.4에서 Km치는 $0.6{\times}10^{-5}M$이였다. 전극의 안정성은 연주일 이상 측정이 가능하였으며, 정량 가능한 범위는 $5.5{\times}10^{-5}M∼2.5{\times}10^{-2}M$ 이며, 감응도는 53.7mv/decade이고, 측 정소요시간은 16~18분이 소요되였다. 방해물질로서 각종 염의 영향은 크게 받지 않음을 알았다. 본 연구에서 제작한 조직 biosensor는 효소분리과정을 거치지 않고 조직을 이용하여 malate흘 정량하였다는 것과 지금까지 연구된 효소 바이오센셔와는 랄리 강산성에셔 측정하던가, 한 가지 이상의 효소를 사용하지 않고도 중성영역인 pH=8에서 측정이 가능 하였으며, malate를 탈수소화하여 정량한 연구는 많았지만, 탈탄산화하여 정량한 연구는 없었다. 본 연구에서 제작한 조직바이오센셔는 malate 측정용 sensor로 가능하리라 생각된다.

• 한국식품과학회지
• /
• 제16권1호
• /
• pp.90-94
• /
• 1984

Malo-alcohol 발효효모(醱酵酵母) Schizosaccharomyces japonicus var. japonius St-3에 의한 사과산의 대사경로(代謝經路)를 검토한 결과는 다음과 같다. Schiz, japonicus var. japonicus St-3의 조효소(粗酵素)의 활성(活性)을 측정(測定)한 바, malic enzyme(EC1. 1. 1. 40)의 효소활성(活性)이 malate dehydrogenase(EC1. 1. 1. 37)보다 약 4배 높았으며 양효소(兩酵素)의 기질(基質)인 L-malate에 대한 Km치(値)는 malic enzymed l 3. 125mM, malated dehydrogenase는 4. 761mM로써 기질에 대한 친화성(親和性)에 있어서 malic enzyme 쪽이 훨씬 컸다. malo-alcohol발효(醱酵)과정중 사과산이 소시(消矢)되어 pyruvate가 생성(生成)됨을 확인(確認)할 수있었으며 $Mn^{2+}$에 의해 malic enzyme의 활성(活性)이 촉진(促進)되었다. 이상의 결과(結果)로 미루어 본(本) 공시균(供試菌)에 의한 사과산 대사(代謝)의 주경로(主經路)는 $malate{\rightarrow}pyruvate{\rightarrow}acetaldehyde{\rightarrow}ethanol$의 경로(經路)를 거치는 것으로 추정(推定)할 수 있었다.

• Preventive Nutrition and Food Science
• /
• 제4권1호
• /
• pp.79-83
• /
• 1999

A potentiometric biosensor employing a CO3-2 ion-selective electrode(ISE) and malic enzyme immobilization in al flow injection analysis (FIA) system was constructed. Analytical parameters were optimized for L-malate determination . The CO3-2 -ISE-FIA system was composed of a pump, an injector, a malic enzyme (EC1.1.1.40) reactor, a CO3-2 ion-selective electrode, a pH/mV meter and a recorder. Cofactor NADP was also injected with substrate for theenzyme reaction into the system. Optimized analytical parameters for L-malate determination in the CO3-2 ISE-FIA system were as follows ; flow rate, 14.5ml/hr ; sample injection volume, 100ul; enzyme loading in the reactor, 20 units ; length of the enzyme reactor , 7 cm ; tubing length form the enzyme reactor to the detector as a geometric factor in FIA, 15 cm . The response time for measuring the entire L-malate concentration range (10-2 ~10-5 mol/L ; 4 injections )was <15minutes . In this CO3-2 -ISE-FIA system, the potential differences due to th eformation of CO3-2 by the reaction of malic enzyme on L-malate were correlated to L-malate concentration in the range of 10-2 ~10-5mol/L ; the detection limit was 10-5 mol/L. This potentionmetric CO3-2 ISE--FIA system was found to be useful for L-malate measurement.

• Journal of Microbiology and Biotechnology
• /
• 제7권6호
• /
• pp.435-437
• /
• 1997

The role of $Ca^{2+}$ in making functional malate synthase in Corynebacterium glutamicum was investigated using the cloned DNA coding for the enzyme. Introduction of cloned aceB into C. glutamicum overexpressed malate synthase as judged by SDS-PAGE. However, the increase in enzyme activity of the expressed malate synthase did not match the level of overexpression observed in SDS-PAGE. Addition of $Ca^{2+}$ to the growth medium specifically increased the activity. The malate synthase could be stained with ruthenium red in a $Ca^{2+}$-specific manner. This agrees with the previous observation which reported a potential $Ca^{2+}$-binding domain in the N-terminal region of the protein.

• 한국약용작물학회지
• /
• 제16권4호
• /
• pp.261-267
• /
• 2008

Malate dehydrogenase is a ubiquitous enzyme in plants, involving in a range of metabolic processes depending on its subcellular location. A malate dehydrogenase (PgMDH) cDNA was isolated and characterized from the root of Panax ginseng C. A. Meyer. The deduced amino acid sequence of PgMDH showed high similarity with the NAD-dependent mitochondrial malate dehydrogenase from Glycinemax (P17783), Eucalyptus gunnii (P46487), and Lycopersicon esculentum (AAU29198). And the segment of a malate dehydrogenase gene was amplified through RT-PCR. The expression of PgMDH was increased after treatments of chilling, salt, UV, cadmium or copper treatment.

• Journal of Animal Science and Technology
• /
• 제51권6호
• /
• pp.511-520
• /
• 2009

The purpose of this study was to investigate effects of increased levels of eugenol, thymol and malate on pH and the concentrations of VFA, lactate and ammonia-N during in vitro ruminal incubation. One Hanwoo beef steer (741 kg) fitted with a rumen cannula was used and fed 0.5 kg/day rice straw and 10 kg/day corn-based concentrate (ratio of concentrate to rice straw = 95 : 5 on DM basis). Three different doses of thymol, eugenol and malate were used. Treatments of the experiment were as follows: Treatments of thymol were control (1g D-glucose/40ml), T1 (1g D-glucose + 40 mg thymol/40 ml), T2 (1g D-glucose + 50 mg thymol/40 ml) and T3 (1g D-glucose + 60 mg thymol/40 ml). Treatments of eugenol were control (1g D-glucose/40 ml), E1 (1g D-glucose + 55 mg eugenol/40 ml), E2 (1g D-glucose + 65 mg eugenol/40 ml) and E3 (1g D-glucose + 75 mg eugenol/40 ml). Treatments of malate were control (1g D-glucose/40ml), M1 (1g D-glucose + 25 mg malate/40ml), M2 (1g D-glucose + 50 mg malate/40 ml) and M3 (1g D-glucose + 100 mg malate/40 ml). The results of this study showed that eugenol and thymol have improved stability of the ruminal fermentation by decreasing lactic acid concentration and increasing ruminal pH. However, it inhibited the production of total VFA, acetate and propionate. Malate also improved stability of the ruminal fermentation by decreasing lactic acid concentration and increasing ruminal pH, but it had a very little effect on ruminal lactate concentrations and pH. On the other hand, malate did not decrease the concentrations of total VFA, acetate and propionate. Therefore, at the low ruminal pH expected in high-concentrate diets, thymol, eugenol, and malate are potentially useful in Hanwoo finishing diets. Further studies are necessary for determining the effectiveness of these additives on in vivo rumen fermentation and animal performance in Hanwoo finishing steers.

• Asian-Australasian Journal of Animal Sciences
• /
• 제19권3호
• /
• pp.368-375
• /
• 2006

Four rumen-fistulated dairy steers were randomly assigned according to a $4{\times}4$ Latin square design to investigate effects of supplementation levels of sodium dl-malate in concentrates on rumen ecology, ruminal fermentation, nitrogen balance, feed intake and digestibility of nutrients and ruminal microbial protein synthesis. The dietary treatments were cassava concentrate-based, containing sodium dl-malate supplementation at 0, 9, 18 and 27 g/hd/d with urea-treated rice straw (UTS) fed ad libitum. The experiment was conducted for four periods, each period lasting 21 days. Ruminal pH increased with incremental addition of malate (p<0.05). Additionally, molar proportions of propionate were higher in supplemented groups and was highest at 18 g/hd/d of malate supplement (p<0.05). Microbial protein synthesis tended to be higher in dairy steers receiving sodium dl-malate supplements and also was the highest at 18 g/hd/d. Variable bacterial populations, such as amylolytic, proteolytic and cellulolytic species were increased (p<0.05). Furthermore, protozoal populations were decreased significantly (p<0.05), while fungal zoospores were dramatically increased in dairy steers receiving sodium dl-malate supplement (p<0.05). These results suggested that supplementation of concentrate containing a high level of cassava chip at 18 g/hd/d with UTS in dairy steers could improve rumen fermentation efficiency and rumen microbial protein synthesis.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제18권2호
• /
• pp.91-95
• /
• 2009

The presence of considerable amount of enzymes of TCA cycle isocitrate dehydrogenase (ICDH-NADP+, EC1.1.1.42), $\alpha$-ketogluterate dehydrogenase ($\alpha$-KGD, EC1.2.4.2) and malate dehydrogenase (MDH, EC1.1.1.37) in fresh control and in vitro starved adult Isoparorchis hypselobagri establish the functional TCA cycle in this fluke. The major metabolic end products are pyruvate, lactate, oxaloacetate and malate. The ratio of oxaloacetate/malate assess that oxaloacetate is reduced to malate and in this fluke the reverse TCA cycle is active. The pyruvate/lactate ratio shows pyruvate is reduced to lactate and the fluke is homolactate farmenters.

• Journal of Plant Biology
• /
• 제22권3호
• /
• pp.55-57
• /
• 1979

These studies were undertaken to determine the $CO^2$fixation patterns following the chloroplast development in maize leaves. At the early stage of chloroplast development $^{14}C$ was incorporated into aspartate (41%) and malate (22%) respectively. Whereas the incorporation of $^{14}C$ into malate was higher than that of aspartate as chloroplast developed. Activity of NADPH-dependent malate dehydrogenase was increased throughout chloroplast development, but that of aspartate transaminase was not. Much incorporation of $^{14}C$ into aspartate at the early stage of chloroplast development and into malate at later stage of chloroplast development lead us to conclude that NADPH-dependent malate dehydrogenase activity is closely associated with chloroplast development, but activity of aspartate transaminase is not.