• 제목/요약/키워드: $_L$-LDH

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Cloning and Characterization of the $_L$-Lactate Dehydrogenase Gene (IdhL) from Lactobacillus reuteri ATCC 55739

  • Park, Jar-Yong;Park, Sun-Jung;Nam, Su-Jin;Ha, Yeong-Lae;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제12권5호
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    • pp.716-721
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    • 2002
  • The ldhL gene encoding the $_L$-(+) lactate dehydrogenase was cloned from Lactobacillus reuteri ATCC 55739 chromosomal DNA and characterized. An internal 750-bp fiagment of ldhL gene was amplified by PCR using primers based on the conserved region of lactobacilli ldhL genes. A genomic library off. reuteri ATCC 55739 was constructed using pBR322, and colony hybridization experiments were performed using the 750-bp fragment as aprobe. One clone harboring a 4.0-kb PstI fragment was identified, and nucleotide sequencing confirmed it as an open reading frame of 972 bp in size in the middle. In addition to IdhL gene, an ORF homologous to Streptococcus pneumoniae TIGR4 hydrolase gene and 3' part of phosphomevalonate kinase gene (mvaK2) were also found on the 4 kb fragment. $_L$-LDH of L. reuteri ATCC 55739 showed the highest degree of homology with the $_L$-LDH of Pediococcus acidilactici (62.4%), fullowed by the $_L$-LDH of Lactobacillus pentosus (58.7%). The size of IdhL transcript determined by Northern blot was 1 kb, indicating the monocistronic nature of IdhL.

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.

國內詞育 원숭이의 血淸 LDH의 總活性値와 isoenzyme에 관한 硏究 (Studies on the Serum Total Activities and Isoenzyme PAtterns of LDH in Non-Human Primates Reared in Korea)

  • 윤상보;김덕환;서지민;신남식;현병화;김명철;윤효인;박배근;송희종
    • 한국임상수의학회지
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    • 제18권4호
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    • pp.380-389
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    • 2001
  • Non-human primates have been increasing in demand as important experimental animals and companion animals, domestically and internationally. The number of non-human primates for these purposes will be much enhanced in the near future. Despite this trend, basic physiological data are scarcely available in these animal species, leading to the difficulty to diagnose diseases when necessary, due to the absence of reference values. Particularly, there is not any report on the total activity of LDH of non-human primates, let alone LDH isoenzyme patterns, in Korea. LDH isoenzymes have a high level of efficaciousness as diagnostic and prognostic aids in various diseases. In this study, total activities and isoenzyme patterns of LDH were measured to obtain their reference values in domestically reared common marmosets, crab-eating macaques and Japanese macaques. There were widespread different values of serum total LDH among the non-human primate species experimented in this study. Serum LDH values of common marmosets and crab-eating macaques were 597.5$\pm$243.1 IU/l and 605.3$\pm$312.6 IU/l, respectively, whereas those of Japanese macaque showed 1,209$\pm$473.8 IU/l. Five isoenzyme fractions of LDH were observed in all experimented non-human primates but their ranks and proportions represented different patterns one another. In common marmosets, the percent of fraction for serum LDH1, LDH$_2$, LDH$_3$, LDH$_4$, and LDH$_{5}$ was 13.7$\pm$6.4%, 23.3$\pm$3.6%, 29.2$\pm$5.0%, 9.4$\pm$1.4% and 24.4$\pm$7.5%, respectively. The rank of LDH isoenzymes was LDH$_3$>LDH$_{5}$>LDH$_2$>LDH$_1$>LDH$_4$, in the descending order. For crab-eating macaques, the fraction of serum LDH$_1$, LDH$_2$, LDH$_3$, LDH$_4$, and LDH$_{5}$ occupied 19.5$\pm$12.7%, 25.3$\pm$9.3%, 23.8$\pm$8.1%, 10.2$\pm$2.8% and 21.3$\pm$14.2%, respectively. The order of LDH isoenzymes was LDH$_2$>LDH$_3$>LDH$_{5}$>LDH$_1$>LDH$_4$, from top to down. On the while, in Japanese macaques, the fraction of serum LDH$_1$ to LDH$_{5}$ showed 23.4$\pm$11.8%, 30.5$\pm$4.1%, 17.4$\pm$3.9%, 11.3$\pm$3.7% and 13.8$\pm$5.6%, respectively. The decreasing order indicated LDH$_2$>LDH$_1$>LDH$_3$>LDH$_{5}$>LDH$_4$. In conclusion, values such as LDH and LDH isoenzyme patterns of investigated for the first time from non-human primates reaared in Korea, could be reference values for the optimal diagnosis and therapy of diseases of the corresponding animal species. Other parameters of hematology and blood biochemistry are urgently needed to study for the benefit of our intimate non-human primates.an primates.

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상부요추와 하부요추 단분절 추간판 탈출증 환자의 임상적 특성과 한방치료 효과 비교연구 (Comparative Study on the Clinical Characteristics and Effects of Korean Medical Treatment between the Upper and Lower Lumbar Single Level Disc Herniation)

  • 이주영;김용현;김광휘;김태연;이태걸;이상운;추희영;정희경;정범환
    • 한방재활의학과학회지
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    • 제28권4호
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    • pp.43-50
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    • 2018
  • Objectives Upper lumbar disc herniation (LDH) (L1/2, L2/3) has specific anatomical characteristics and different outcome after conventional treatment compared to lower LDH (L3/4, L4/5, L5/S1). The purpose of this study was to compare the clinical features and effects of korean medical treatment of upper LDH between lower LDH. Methods We retrospectively reviewed the clinical data collected from 121 patients who was had admitted at the Haeundae Jaseng Hospital of Korean Medicine from June 1st, 2014 to August 31th, 2018. The patients who had treated at L1/2, L2/3 level LDH were grouped and compared with those treated at the L3/4, L4/5, L5/S1 level. We reviewed the patient characteristics such as age, the positive rate of Straight Leg Raise Test (SLR test), the presence or absence of previous lumbar surgery. Numeric Rating Scale (NRS) and Oswestry Disability Index (ODI) were evaluated at adimission and discharge. Results Mean age was significantly higher at upper LDH group and positive rate of SLR test was higher at lower LDH group. There was no significant difference of gender and previous lumbar surgery between two groups. Each group had a significant improvement of NRS, ODI scores. But between two groups, there was no significant difference of NRS, ODI scores. Conclusions Upper LDH group was older than lower LDH group. SLR test was useful tool to exclude upper LDH. Korean medical treatment was significantly effective to both upper and lower LDH groups. Further well designed prospective comparative studies are needed.

Regulation of Metabolic Flux in Lactobacillus casei for Lactic Acid Production by Overexpressed ldhL Gene with Two-Stage Oxygen Supply Strategy

  • Ge, Xiang-Yang;Xu, Yan;Chen, Xiang;Zhang, Long-Yun
    • Journal of Microbiology and Biotechnology
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    • 제25권1호
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    • pp.81-88
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    • 2015
  • This study describes a novel strategy to regulate the metabolic flux for lactic acid production in Lactobacillus casei. The ldhL gene encoding L-lactate dehydrogenase (L-LDH) was overexpressed in L. casei, and a two-stage oxygen supply strategy (TOS) that maintained a medium oxygen supply level during the early fermentation phase, and a low oxygen supply level in the later phase was carried out. As a consequence, a maximum L-LDH activity of 95.6 U/ml was obtained in the recombinant strain, which was over 4-fold higher than that of the initial strain. Under the TOS for L. casei (pMG-ldhL), the maximum lactic acid concentration of 159.6 g/l was obtained in 36 h, corresponding to a 62.8% increase. The results presented here provide a novel way to regulate the metabolic flux of L. casei for lactic acid production in different fermentation stages, which is available to enhance organic acid production in other strains.

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.

흉부수술 환자에서의 CPK 와 LDH I soenzyme 의 변화 (Comparison of serum isoenzyme levels of CPK and LDH in patients ungergoing thoracic operations)

  • 이영욱
    • Journal of Chest Surgery
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    • 제15권4호
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    • pp.460-466
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    • 1982
  • A analysis of CPK & LDH Isoenzyme was done on the consecutive patients undergoing thoracic operations from July 1982 to October 1982 in the Department of Thoracic and Cardiovascular Surgery, Capital Armed Forces General Hospital. Eighteen patients were analysed by three groups, such as open heart surgery [group A], major thoracic operation [group B] , minor thoracic operation group [group C]. In all patients serial determination of total level and Isoenzyme of CPK, LDH wad done on preoperative operative and up to 8th post-operative day, The results obtained are as follows. 1. The average value of serum CPK before the operation was 61 IU/L. The value of serum CPK was increased following the operation mainly MM portion and reached to the maximal level of 536107 IU/L in A group 1200191 IU/L in B group, 306150 IU/L in C group on the first postoperative day. The enzyme activity was gradually decreased thereafter and returned to the normal range on the 3rd or 4th day after the operation. 2. The average value of serum LDH before the operation was found to be 83 IU/L. The value was increased during the operation and reached to the maximal level of 481108 IU/L in group A, 14827 I U/L in group B, 10035 IU/L in group C on the second day after the operation. The enzyme activity was gradually decreased thereafter and returned to the normal range on the seventh day after the operation. The enzyme activity was dependent to the duration of operation, severity of muscle damage, type of thoracotomy, effect of extracorporeal circulation, state of disease.

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칠성장어(Lampetra japnica) 간조직 젖산탈수소효소와 대구(Gadus macrocephalus) liver-Specific C4동위효소의 특성 및 진화적 관계 (Characterization and Evolutionary Relationship of Lactate Dehydrogenase in Liver of Lampetra japonica and Liver-specific C4 Isozyme in Gadus macrocephdus.)

  • 박선영;조성규;염정주
    • 생명과학회지
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    • 제14권4호
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    • pp.708-715
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    • 2004
  • 칠성장어(Lampetra japonica) 간조직 젖산탈수소효소(EC 1.1.1.27, lactate dehydrogenase, LDH) 동위효소는 affinity chromatography에서 buffer를 유입한 후 용출된 분획에서 정제되었다. 대구(Gadus macrocephalus)의 liver-specific $C_4$동위효소는 열처리한 후 affinity chromatography하여 NAD+ 를 함유한 buffer에서 용출되기 시작하여 buffer를 유입한 후 $B_4$ 동위효소와 함께 용출되어, DEAE-Sephacel chromatography에 의해 정제되었다. 대구 간조직에서 열에 대한 안정성은$C_4$$B_4$$A_4$ 동위효소의 순서로 나타났다. Chromate-focusing에 의해 정제한 칠성장어 간조직의 pH 7.45 분획의 LDH 동위효소는 정제된 간조직 LDH보다 피루브산에 의한 기질저해도가 컸다. 칠성장어 간조직 LDH의 최적 pH는 7.5, liver-specific $C_4$동위효소는 pH 8.5였다. 칠성장어 간조직 LDH는 항원-항체반응에서 꺽지 $A_4$ 항체와 liver-specific $C_4$ 항체의 순서로 반응하였고 eye-specific $C_4$ 항체와는 반응 정도가 낮았다. 따라서 칠성장어 간조직 LDH는 하부단위체 A와 liver-specific $C_4$의 구조와 유사하게 진화되었으며, 하부단위체 C 는 진화속도가 매우 빠른 것으로 확인되었다. 칠성장어 간조직의 LDH는 단일 동위효소가 아니라, 하부단위체 A, B 및 C로 구성된 동위효소들인 것으로 사료된다.

기아상태에서 Ldh-C가 발현된 어류 조직의 젖산탈수소효소의 대사 (Metabolism of Lactate Dehydrogenase in Tissues from Ldh-C Expressed Fish at Starved State)

  • 염정주;김규동
    • 생명과학회지
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    • 제26권2호
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    • pp.155-163
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    • 2016
  • 젖산탈수소효소(Lactate dehydrogenase, EC 1.1.1.27, LDH) LDH-C의 기능을 확인하기 위해 liver-specific Ldh-C가 발현된 붕어(Carassius auratus)와 eye-specific Ldh-C가 발현된 파랑볼우럭(Lepomis macrochirus)을 기아 상태로 유지시킨 후(S) 조직들의 LDH 대사를 연구하였다. 기아 후 붕어 간조직의 LDH 활성이 크게 증가되었으며 LDH 비활성(units/mg)과 LDH/CS는 조직들에서 증가되어 혐기적 대사가 이루어짐을 확인하였다. 기아 후 LDH B4 동위효소가 골격근조직에서 감소되었고 심장조직에서 증가되었다. 눈과 뇌조직에 나타났던 LDH C4 동위효소는 liver-specific C4로 확인되었으며 기아 후에 없어지고, 눈조직은 C hybrid, 뇌조직은 A4, 간조직은 C hybrid와 C4 동위효소가 각각 증가되었다. 그러나 파랑볼우럭 조직에서 LDH 활성의 변화는 작았으나 눈조직에서 가장 크게 증가되었으며, 뇌조직은 LDH A4와 AC hybrid가 증가되었다. 피루브산 10 mM에 의해 기아 후 붕어 조직의 LDH 활성은 30.30-18.64%, 파랑볼우럭 조직의 LDH는 25-18.75% 남았으며, 붕어는 KmPYR 값이 증가되었다. 실험 결과 LDH liver-specific C 동위효소가 기아 중에 간, 뇌 및 눈조직에서 발현되었고, 기아 후 뇌조직에서 젖산의 대사가 우세하고, 붕어 LDH liver-specific C가 파랑볼우럭 LDH eye-specific C보다 영향을 더 받는 것으로 사료된다.

젖산탈수소효소 eye-specific C4 동위효소의 생화학적 특성: 파랑볼우럭(Lepomis macrochirus)과 큰입우럭(Micropterus salmoides) (Biochemical Properties of Lactate Dehydrogenase Eye-Specific C4 Isozyme: Lepomis macrochirus and Micropterus salmoides)

  • 염정주;구보라
    • 생명과학회지
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    • 제22권2호
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    • pp.209-219
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    • 2012
  • 눈조직의 젖산탈수소효소(LDH, EC 1.1.1.27) eye-specific $C_4$ 동위효소의 특성을 native-PAGE, Western blotting, 면역침강반응 및 효소 역학을 이용하여 연구하였고, LDH eye-specific $C_4$ 동위효소의 활성을 측정하는 조건을 제시하였다. 파랑볼우럭(Lepomis macrochirus)과 큰입우럭(Micropterus salmoides) 눈조직의 세포기질에서 LDH eye-specific $C_4$ 동위효소가 확인되었으며 $B_4$ 동위효소보다 $A_4$ 동위효소에 유사하였다. 파랑볼우럭 눈조직의 LDH/CS는 9월에 증가하여 혐기적 대사가 높게 이루어졌다. 파랑볼우럭과 큰입우럭 눈조직 미토콘드리아 LDH의 전기영동상은 세포기질 LDH와 유사하였다. 눈조직의 LDH eye-specific $C_4$ 동위효소는 Preparative native-PAGE에 의해 정제되었다. 파랑볼우럭과 큰입우럭의 LDH eye-specific $C_4$ 동위효소의 활성은 피루브산 0.2 mM과 0.1 mM 이상의 농도에서 각각 감소되었고, 피루브산 10 mM에서 5.2%, 15.8% 활성이 남아 저해 정도가 크게 나타났다. 그리고 눈조직의 LDH 활성도 젖산 22 mM과 24 mM 이상의 농도에서 각각 감소되어졌다. 파랑 볼우럭 eye-specific $C_4$ 동위효소의 $Km^{PYR}$는 0.088 mM, 큰입우럭 eye-specific $C_4$ $Km^{PYR}$는 0.033 mM였으며, 세포기질과 미토콘드리아 eye-specific $C_4$ 동위효소는 ${\alpha}$-ketobutyric acid에서 활성이 높게 나타났다. 눈조직과 eye-specific $C_4$ 동위효소의 활성은 피루브산 0.5 mM과 Tris-HCl 완충액, pH 7.5에서 측정하는 것이 적합한 것으로 확인되었다. 실험 결과, 큰입우럭 LDH eye-specific $C_4$ 동위효소는 파랑볼우럭 LDH eye-specific $C_4$보다 피루브산에 대한 친화력이 큰 것으로 확인되었고, 더 낮은 피루브산의 농도에서 최대 활성에 이르고, 더 높은 젖산의 농도에서 최대 활성을 나타냈다. 따라서 LDH eye-specific $C_4$ 동위효소에 의해 생성된 에너지가 포식 행동의 초기 반응에 사용되는 것으로 사료된다.