• 제목/요약/키워드: Dehydrogenases

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2,3,5-Triphenyltetrazolium Chloride를 이용한 병원성 미생물 확인시험에 관한 연구 (Study of 2,3,5-Triphenyltetrazolium Chloride for Detection of Pathogenic Microorganisms)

  • 강정욱;배준태;연재영;김영호;김진화;이근수;표형배
    • 대한화장품학회지
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    • 제40권3호
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    • pp.307-311
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    • 2014
  • 2,3,5-Triphenyltetrazolium chloride (TTC)는 산화환원지시약으로 미생물의 증식에 의한 산소 소비를 쉽게 확인할 수 있다. 용해 후 무색의 형태를 띠고 있으나 생리활성이 있는 조직에서는 탈수소 효소(dehydrogenases)에 의해 환원되어 빨간색의 불용성 1,3,5,-triphenylformazan (TPF)가 된다. 본 연구에서는 병원성 미생물(Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecium, Candida albicans)에 TTC 지시약을 활용하여 미생물 성장시험에 대해 확인하였다. 시험 균주에 TTC를 첨가하여 확인한 결과, 모두 탈수소효소 반응으로 인한 TPF 형성으로 붉은색 콜로니를 관찰하였다. 이후 TTC 0.04% 이상의 농도 및 12 h 이상 배양조건으로 최적화 실험 후 균주별 CFU 값을 통해 TPF 발현능을 확인하였다. 결국 TTC가 병원성 세균 및 효모균 성장에 큰 영향을 끼치지 않으며 배양 시 세균의 경우 12 h, 효모균의 경우 48 h 이후부터 확인이 가능하였다. 이러한 결과들로부터 TTC를 활용한 미생물 성장 확인 시험법이 더 신속 정확한 방법으로 화장품 연구에 활용 가능할 것으로 사료된다.

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

  • 장명운;박정미;이홍균;이소라;김태집
    • 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.

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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    • 제9권4호
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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.

Upregulation of Glutathion S-Transferase mu 1 in Bovine Cystic Follicles

  • Kang, Da-Won;Kim, Chang-Woon;Han, Jae-Hee
    • 한국수정란이식학회지
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    • 제25권4호
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    • pp.273-279
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    • 2010
  • Follicular cystic follicles (FCFs) show delayed regression with persistent follicle growth. However, the mechanism by which follicles are persistently grown remains unclear. Glutathione S-transferases (GSTs) are drug-metabolizing and detoxification enzymes that are involved in the intracellular transport and metabolism of steroid hormones. In this study, a proteomic analysis was performed to identify whether GST expression is changed in bovine FCFs and to predict the interactions between GST and other proteins. Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and 25 mm). In bovine follicles, GST mu 1 (GSTM1) was detected as a differentially expressed protein (DEP) and significantly up-regulated in FCFs compared to normal follicles (p<0.05). Consistent with the proteomic results, semi-quantitative PCR data and western blot analysis revealed an up-regulation of GSTM1 in FCFs. Expression levels of aromatase and dehydrogenase proteins were changed in FCFs. These results show that the up-regulation of GSTM1 that is observed in bovine FCFs is likely to be responsible for the persistent follicle growth in FCFs as the activity of aromatase and the dehydrogenases.

兩棲類 胚發生에 EK른 Lactate Dehydrogenase 및 Malate Dehydrogenase의 Isozyme 변화에 관하여 (Studies on the Change of Isozyme Patterns of Lactate and Malate Dehydrogenases During Embryonic Development of Some Amphibians)

  • Park, Young-Chul;Ha, Doo-Bong
    • 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.

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계배 대뇌의 신경세포 분화에 따른 탈수소효소 활성 및 미세구조 (1) (Ultrastructure and Dehydrogenase activity on the Differentiation of the Cerebral Nerve Cell in the Chick Embryo (1))

  • 김생곤
    • Applied Microscopy
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    • 제28권4호
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    • pp.563-575
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    • 1998
  • To investigate the changes during the differentiation of the cerebral neurons of chick embryo of tne embryogenic day (ED) 7 and 8, the ultrastructural changes in the cerebral neurons, the activity of dehydronases (LDH, MDH and SDH), protein expression profile and adenosine triphosphate concentration were analyzed. In ED 7 chick embryos, relatively large nucleus, centrally located nucleolus, evenly spread chromatin over nucleoplasm, and prominent nuclear envelope were observed. Oval-shaped mitochondria with well-developed cristae were present over entire cytoplasm. In ED 8 chick embryos, evenly spread chromatin over nucleoplasm, and prominent nuclear envelope were observed. In the cytoplasm, well-developed rough endoplasmic reticulum and Golgi complex were observed. In ED 7 chick embryos and ED 8 chick embryos, 31 polypeptide bands and 34 polypeptide bands were observed, respectively. The activities of dehydrogenases were lower in ED 7 chick embryos than in ED 8 chick embryos. LDH activity was 8.16 (ED 7) and 9.28 (ED 8), MDH activity was 7.98 (ED 7) and 10.10 (ED 8), and SDH activity was 5.49 (ED 7) and 7.14 (ED 8) respectively. The ATP concentration remained unchanged over ED 7 and 8.

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Retinoid Metabolism in the Degeneration of Pten-Deficient Mouse Retinal Pigment Epithelium

  • Kim, You-Joung;Park, Sooyeon;Ha, Taejeong;Kim, Seungbeom;Lim, Soyeon;You, Han;Kim, Jin Woo
    • Molecules and Cells
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    • 제44권8호
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    • pp.613-622
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    • 2021
  • In vertebrate eyes, the retinal pigment epithelium (RPE) provides structural and functional homeostasis to the retina. The RPE takes up retinol (ROL) to be dehydrogenated and isomerized to 11-cis-retinaldehyde (11-cis-RAL), which is a functional photopigment in mammalian photoreceptors. As excessive ROL is toxic, the RPE must also establish mechanisms to protect against ROL toxicity. Here, we found that the levels of retinol dehydrogenases (RDHs) are commonly decreased in phosphatase tensin homolog (Pten)-deficient mouse RPE, which degenerates due to elevated ROL and that can be rescued by feeding a ROL-free diet. We also identified that RDH gene expression is regulated by forkhead box O (FOXO) transcription factors, which are inactivated by hyperactive Akt in the Pten-deficient mouse RPE. Together, our findings suggest that a homeostatic pathway comprising PTEN, FOXO, and RDH can protect the RPE from ROL toxicity.

cDNA Sequence and mRNA Expression of a Putative Alcohol Dehydrogenase from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Lee, Young-Sin;Ryu, Kang-Sun
    • International Journal of Industrial Entomology and Biomaterials
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    • 제7권1호
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    • pp.37-44
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    • 2003
  • Alcohol dehydrogenases (AHDs) are enzymes responsible for the catalysis of the reversible conversion of various alcohols to their corresponding aldehydes and ketonesis. Until now cDNA sequences of ADH gene is informed exclusively from several diptean species. We describe here the cDNA sequence and mRNA expression of a putative ADH gene from the mole cricket, Gryllotalpa orientalis, and phylogenetic relationships among known insect ADHs. The G. orientalis ADH cDNA sequences comprised of 798 bp encoding 266 amino acid residues. The multiple sequence alignment of G. orientalis ADH gene and known dipteran ADHs shared 100% identity in the nine amino acid residues that are important for the enzymatic activity in Drosophila melanogaster. Percent sequence identity ranged from 25% to 32% among all insect ADHs including both types of ADHs. G. orientalis ADH gene showed no clear resemblance to any dipteran species and type. Phylogenetic analysis of the deduced amino acid sequences of G. orientalis ADH gene with available dipteran ADH genes including both types of ADHs further confirmed that the G. orientalis ADH gene is not clearly assigned to either type of ADHs. Northern blot analysis revealed a stronger signal in the fat body than midgut and epidermis, indicating that the fat body possibly is a main site for the synthesis of the G. orientalis ADH protein.

Proteomic Analysis of Proteins Increased or Reduced by Ethanol of Lactobacillus plantarum ST4 Isolated from Makgeolli, Traditional Korean Rice Wine

  • Lee, Seung-Gyu;Lee, Kang-Wook;Park, Tae-Heung;Park, Ji-Yeong;Han, Nam-Soo;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제22권4호
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    • pp.516-525
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    • 2012
  • LAB were isolated from makgeolli locally produced around Jinju, Gyeongnam, S. Korea during spring of 2011. Randomly selected 11 isolates from MRS agar plates were identified first by API CHL 50 kits and then 16S rRNA gene sequencing. All 11 isolates were identified as Lactobacillus plantarum. Among them, ST4 grew in MRS broth with ethanol up to 10%, showing the highest alcohol resistance. L. plantarum ST4 was moderately resistant against acid and bile salts. When cellular proteins of L. plantarum ST4 under ethanol stress were analyzed by two-dimensional gel electrophoresis (2DE), the intensities of 6 spots increased, whereas 22 spots decreased at least 2-fold. Those 28 spots were identified by peptide mass fingerprinting (PMF). FusA2 (elongation factor G) increased 18.8-fold (6% ethanol) compared with control. Other proteins were AtpD (ATP synthase subunit beta), DnaK, GroEL, Tuf (elongation factor Tu), and Npr2 (NADH peroxidase), respectively. Among the 22 proteins decreased in intensities, lactate dehydrogenases (LdhD and LdhL1) were included.