• 한국미생물·생명공학회지
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• 제27권2호
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• pp.124-128
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• 1999

TK6 was able to produce NAD+ dependent cyclohexanol dehydrogenase(CDH). The production of CDH was increased rapidly at the logarithmic phase and maintained constantly after that. In order to investigate the inductive production of CDH by various substrates, the bacteria were grown in the media containing alicyclic hydrocarbons and various alcohols as a sole crabon souce. CDH was induced most actively by cyclohexanol. Cyclohexanone and cyclohexane-1,2-diol also induced remarkable amount of CDH but it was induced weakly by 1-propanol, 1-butanol, 1-pentanol, 1-hexanol, 2-propanol, and 2-methyl-1-propanol. The dehydrogenase of the bacteria grown in the media containing cyclohexanol were weakly active for various alcohols, but the dehydrogenase activity for cyclohexane-1,2-diol was twice as much as that for cyclohexanol. Activity staining on PAGE of the cell free extract of Rhodococcus sp. TK6 grown in the media containing cyclohexanol reveals at least sever isozyme bands of CDH and we nominated the four major activity bands as CDH I, II, III, and IV. CDH I was strongly induced by cyclohexanol, cyclohexane-1,2-diok, but its activity was specific to cyclohexane-1,2-diol and 1-pentanol. CDH IV was strongly induced by cyclohexanol and cyclohexane-1,2-diol, and its activity was very specific to cyclohexane-1,2-diol.

• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.197-201
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• 2004

A 2-D gel protein analysis of Lactobacillus reuteri ATCC 55739 produced spots corresponding to subunits of the pyruvate dehydrogenase complex, as identified by N-terminal protein sequencing. Oligonucleotide probes specific for the subunits of the pyruvate dehydrogenase complex were synthesized ,md used to screen a L. reuteri genomic library to clone the structural genes. Two positive clones were isolated and identified as having the same 2.2 kb insert. A pdhB encoding the $\beta$-subunit of El subunit (pyruvate dehydrogenase component) of the pyruvate dehydrogenase complex was located in the middle of the insert. Furthermore, a 5' truncated pdhA encoding the $\alpha$-subunit of the E1 subunit and a 3' truncated pdhC encoding the E2 subunit (dihydrolipoamide acetyltransferase) were also located upstream and downstream of the pdhB, respectively.

• 한국축산식품학회지
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• 제24권2호
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• pp.151-155
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• 2004

본 연구는 미토콘드리아 malate dehydrogenase활성을 이용하여 냉장계육과 냉동계육의 판별법을 개발하기 위하여 실시되었다. 단백질의 함량은 8.5mg/mL에서 12.7mg/mL 범위로 나타났으며, 동일 저장 기간 동안의 온도 별 차이점은 모든 부위에서 1일에서 15일 저장기간이 길어짐에 따라 단백질함량이 늘어나는 경향을 보인다. 냉장육의 보수력은 저장 기간 1일 60.5%에서 15일 33.9%로 감소하였으며 유의적 차이는 없었다(p＞0.05). 냉장계육과 냉동계육은 저장 기간이 길어짐에 따라 육즙손실량이 증가하였으며 유의적 차이는 없었다.(p＞0.05). 냉장계육의 육즙손실량이 냉동계육의 육즙손실량보다 낮게 나타났다. 냉장계육과 냉동계육 간의 mitochondrial malate dehydrogenase의 활성은 냉동계육이 냉장계육보다 효소활성이 높게 나타났다. 따라서 본 연구결과로부터 mitochondrial malate dehydrogenase의 활성 여부로 계육의 냉장, 냉동 유무를 판별하는데 유효한 지표로 사용 가능한 것으로 사료되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권3호
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• pp.421-426
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• 2002

The pyruvate dehydrogenase complex (PDC), a member of $\alpha$-keto acid dehydrogenase complex, catalyzes the oxidative decarboxylation of pyruvate with the formation of $CO_2$, acetyl-CoA, NADH, and $H^+$. This complex contains multiple copies of three catalytic components including pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide dehydrogenase (E3). Two regulatory components (E1-kinase and phospho-E1 phosphatase) and functionally less-understood protein (protein X, E3BP) are also involved in the formation of the complex. In this study, we have partially cloned the gene for E3BP in human. Nine putative clones were isolated by human genomic library screening with 1.35 kb fragment of E3BP cDNA as a probe. For investigation of cloned genes, Southern blot analysis and the construction of the restriction map were performed. One of the isolated clones, E3BP741, has a 3 kb-SacI fragment, which contains 200 bp region matched with E3BP cDNA sequences. The matched DNA sequence encodes the carboxyl-terminal portion of lipoyl-bearing domain and hinge region of human E3BP. Differences between yeast E3BP and mammalian E3BP coupled with the remarkable similarity between mammalian E2 and mammalian E3BP were confirmed from the comparison of the nucleotide sequence and the deduced amino acid sequence in the cloned E3BP. Cloning of human E3BP gene and analysis of the gene structure will facilitate the understanding of the role(s) of E3BP in mammalian PDC.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2002년도 창립10주년기념 및 국립독성연구원 의약품동등성평가부서 신설기념 국재학술대회:생물학적 동등성과 의약품 개발 전략을 위한 국제심포지움
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• pp.236-236
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• 2002

Pyruvate dehydrogenase phosphatase (PDP) is a mitochondrial protein serine/threonine phosphatase that catalyzes the dephosphorylation and concomitant reactivation of the pyruvate dehydrogenase componant of the pyruvate dehydrogenase complex (PDC). PDP consists of a Mg$\^$+2/ -dependent and Ca$\^$+2)-stimulated catalytic subunit (PDPc) of Mr 52,600 and a FAD-containing regulatory subunit (PDPr) of Mr 95.600. Catalytic subunit of pyruvate dehydrogenase phosphatase (PDPc) has been suggested to have three major functional domains such as dihydrolipoamide acetyltransferase(E$_2$)-binding domain, regulatory subunit of PDP(PDPr)-binding domain, and calcium-binding domain. In order to identify functional domains, recombinant catalytic subunit of pyruvate dehydrogenase phosphatase (rPDPc) was expressed in E. coli JM101 and purified to near homogeneity using the unique property of PDPc: PDPm binds to the inner lipoyl domain (L$_2$) of E$_2$ of pyruvate dehydrogenase complex (PDC) in the presence of Ca$\^$+2/, not under EGTA. PDPc was limited-proteolysed by trypsin, chymotrypsin, Arg-C, and elastase at pH7.0 and 30$^{\circ}C$ and N-terminal analysis of the fragment was done. Chymotrypsin, trypsin, and elastase made two major framents: N-terminal large fragment, approx. 50kD and C-terminal small fragment, approx. 0 kDa. Arg-C made three major fragments: N-terminal fragment, approx. 35 kD, and central fragment, approx. 15 kD, and C-terminal fragment, approx. 10 kD. This study strongly suggest that PDPc consists of three major functional domains. However, further study should be necessary to identify the functional role.

• 생명과학회지
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• 제8권1호
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• pp.8-13
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• 1998

옥수수의 미토콘드리아 변이체(NGS2)는 전자전달계 내의 NADH dehydrogenase를 구성하고 있는 subunit 4와 7 유전자의 재조합에 의해서 생성된 변이체이다. 이들의 변이체들은 식물의 성장과 발육에 절대적인 영양을 미치며, 또한 기내에서의 callus line들의 생성과 발달에도 상당한 영향을 미친다. 이들의 미토콘드리아 mutant 들은 3개의 primer를 사용하는 PCR 방법에 의해서 쉽게 선별이 가능하며, 세포 내의 키토콘드리아 변이 정도를 간접적으로 추측케 하며, 체시포 분열시 세포질 내의 기관들이 random으로 분리되는 현상을 간법적으로 알 수 있다. NGS2 mutant들에서 유기된callus line들은 식물체 재문화에도 영향을 미쳐 murant 미토콘드리아가 많은 call line들에슨 실질적인 부정 줄기의 유기를 방해하는 것으로 사료된다. 결론적으로 NADH-dehydrogenase는 식물체가 재분화 또는 성장하는데 있엇 필요한 요소라고 생각된다.

• Biomaterials and Biomechanics in Bioengineering
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• 제3권1호
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• pp.37-46
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• 2016

To maintain activity in a coenzyme/enzyme mixture system, such as ${\beta}$-nicotinamide adenine dinucleotide (NADH)/dehydrogenase, the water-soluble 2-methacryloyloxyethyl phosphorylcholine (MPC) polymers as an additive were synthesized and investigated for their stabilizing function. The inhibitor for the NADH/dehydrogenase reaction was spontaneously formed when the NADH was stored in the dehydrogenase solution. Therefore, we hypothesized that if the additive polymer could interact with an inhibitor without any adverse effect on the dehydrogenase, the activity in the NADH/dehydrogenase mixture could be maintained. We selected lactose dehydrogenase (LDH) as the enzyme, and the NADH was dissolved and incubated at $37^{\circ}C$ in the LDH solution containing the polymers. The phospholipid polymers used in this study were poly(MPC) (PMPC), poly(MPC-co-3-trimethylammonium-2-hydroxypropyl methacrylate chloride) (PMQ) and poly[MPC-co-potassium 3-methacryloyloxypropyl sulfonate ($MSO_3$)] ($PMMSO_3$). The poly($MSO_3$) was used as a reference. For the PMQ and $PMSO_3$ aqueous solutions, the activity of the NADH/LDH mixture system decreased with incubation time as the same level or lower than that in the Tris buffered solution in the absence of the polymers. However, for the poly($MPC-co-MSO_3$) ($PMMSO_3$) aqueous solution, the activity of the NADH/LDH mixed system was six times higher than that in the buffered solution even after a 3-days incubation. The LDH activity was 1.5-1.8 times higher in the presence of the $PMMSO_3$ compared with that in the $PMSO_3$ solution. The mixture of two polymers, poly(MPC) and poly($MSO_3$), did not produce any stabilization. Thus, both the MPC and $MSO_3$ units in the polymer chain had important and cooperative effects for stabilizing the NADH/LDH mixture.

• 미생물학회지
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• 제34권4호
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• pp.207-211
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• 1998

메탄올을 이용하여 성장하는 Methylovorus sp. strain SS1은 formaldehyde의 산화를 위한 linear route의 주효소인 $NAD^+$-linked formaldehyde dehydrogenase 및 $NAD^+$-linked formate dehydrogenas와 cyclic route의 주효소인 hexulose-6-phosphate synthase, glucose-6-phosphate isomerae, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase 등의 활성을 나타내었는데, cyclic route에 관여하는 효소의 활성이 상대적으로 더 높았다. 이 세균은 formaldehyde의 동화와 관련된 ribulose monophosphate 경로의 주효소와 Entner-Doudoroff 경로의 주효소 및 transaldolase 활성과 함께 세포외 다당류 합성과 관련된 phosphoglucomutase, UDP-glucose pyrophyosphorylase, mannose-6-phosphate isomerase의 활성도 나타내었다. 2.3 mM의 ammonium sulfate가 포함된 배지에서 성장한 세균은 7.6 mM의 ammonium sulfate가 포함된 배지에서 성장한 세균보다 더 많은 세포외 다당류를 생산하였지만 균체 수율은 낮았고, 6-phosphogluconate dehydrogenase와 phosphoglucomutase 및 UDP-glucose pyrophoshorylase의 활성은 높게 나타내었으나 6-phosphogluconate dehydratase/2-keto-3-deoxy-6-phosphogluconate aldolase의 활성은 낮았다.

• Bulletin of the Korean Chemical Society
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• 제34권6호
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• pp.1621-1622
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• 2013

• Bulletin of the Korean Chemical Society
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• 제35권6호
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• pp.1603-1604
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• 2014