• Title/Summary/Keyword: lactate dehydrogenase

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Growth Characteristics of a Pyruvate Decarboxylase Mutant Strain of Zymomonas mobilis (Pyruvate decarboxylase 돌연변이 Zymomonas mobilis 균주의 생장 특성 연구)

  • Xun, Zhao;Peter L., Rogers;Kwon, Eilhann E.;Jeong, Sang Chul;Jeon, Young Jae
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1290-1297
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    • 2015
  • Studies of the inactivation of a gene encoding pyruvate decarboxylase, pdc, in an ethanol-producing bacterium, Zymomonas mobilis, identified a mutant strain with 50% reduced PDC activity. To evaluate the possibility of a carbon-flux shift from an ethanol pathway toward higher value fermentation products, including pyruvate, succinate, and lactate, fermentation studies were carried out. Despite attempts to silence pdc expression in the wild-type strain ZM4 using cat-inserted pdc and pdc-deleted homologs by electroporation, the strain isolated showed partial gene activation. Fermentation experiments with the PDC mutant strain showed that the reduced expression level of PDC activity resulted in decreased rates of substrate uptake and ethanol production, together with increased pyruvate accumulation of 2.5 g l-1 , although lactate and succinate concentrations were not significantly enhanced in these modified strains. Despite numerous attempts, no strains were isolated in which complete pdc inactivation occurred. This result indicates that the ethanol fermentation pathway of this bacterium is totally dependent on the activity of the PDC enzyme. To ensure a redox balance of intracellular NAD and NADH levels, other enzymes, such as lactate dehydrogenase for lactate, and enzymes involved in the production of succinic acid, such as pyruvate dehydrogenase (PDH) and malic enzymes, may be needed for their increased end-product production.

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

  • Yum, Jung Joo;Kim, Gyu Dong
    • Journal of Life Science
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    • v.26 no.2
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    • pp.155-163
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    • 2016
  • Metabolism of lactate dehydrogenase (EC 1.1.1.27, LDH) was studied to identify the function of LDH-C. Tissues of LDH liver-specific Ldh-C expressed Carassius auratus and eye-specific Ldh-C expressed Lepomis macrochirus after starvation were studied. LDH activity in liver tissue from C. auratus was increased after starvation. And LDH specific activity (units/mg) and LDH/CS were increased in tissues. It means the anaerobic metabolism was taking place in C. auratus after starvation. LDH B4 isozyme was decreased in skeletal muscle and increased in heart tissue. LDH C4 isozymes those showed in eye and brain tissues were identified as liver-specific C4 isozymes and disappeared after starvation. And C hybrid in eye, A4 isozyme in brain, and both C hybrid and C4 isozyme in liver tissue were increased, respectively. In L. macrochirus, the level of variation of LDH activities was low but greatly increased especially in eye tissue and LDH A4 and AC hybrid were increased in brain tissue. The LDH activities in tissues from C. auratus and L. macrochirus remained 30.30-18.64% and 25-18.75%, respectively, as a result of the inhibition by 10 mM of pyruvate. The KmPYR values of LDH in C. auratus were increased. As a result, LDH liver-specific C4 isozyme was expressed in liver, brain and eye tissues during starvation. It seems metabolism of lactate was predominant in brain tissue. After starvation, the liver-specific LDH-C was affected more than eye-specific LDH-C.

Functional and Physical Interaction between Human Lactate Dehydrogenase B and $Na^+/H^+$ Exchanger Isoform 1

  • Kim, Eun-Hee
    • Animal cells and systems
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    • v.13 no.3
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    • pp.283-288
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    • 2009
  • The ubiquitous plasma membrane $Na^+/H^+$ exchanger 1 (NHE1) is rapidly activated in response to various extracellular stimuli and maintains normal cytoplasmic pH. Yeast two-hybrid screening was used in order to identify proteins interacting with NHE1 using its cytoplasmic domain as a bait from HeLa cDNA library. One of the interacting cDNA clones was human Lactate dehydrogenase B (LDHB). In vitro translated LDHB was pulled down together with GST-NHE1.cd protein in the GST pull down assay, confirming the interaction in vitro. LDHB antibody immunoprecipitated endogenous LDHB together with NHE1 from H9c2 cells, validating cellular interaction between NHE1 and LDHB. Subsequent analysis revealed that the overexpression of LDHB increased intracellular PH, implying opening of the NHE1 transporter. Moreover, overexpression of LDHB activated caspase 3 and induced cell death, consistent with the expected phenotype of hyper-activation of NHE1. Collectively, our data indicate that LDHB modulates NHE1 activity via physical interaction.

Immobilization of enzymes on magnetic nanoparticles for biochip applications

  • Sohn, Ok-Jae;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.692-695
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    • 2003
  • In this work immobilization technique of enzymes onto the magnetic nanoparticles has been developed for biochip applications. Glucose oxidase and lactate dehydrogenase were immobilized on magnetic nanoparticles via cyanamide and glutaraldehyde. Immobilized enzymes had good operational and storage stability The immobilized glucose oxidase and lactate dehydrogenase were characterized by some factors(pH, temperature, and components of buffer solution etc) which affect the activity, In order to characterize the magnetic nanoparticles, we have used transmission electron microscopy(TEM), X-ray diffraction(XRD) and Fourier transform infrared(FTIR).

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Subunit Interactions of Vertebrate Lactate Dehydrogenase: I. Immunochemistry of Subunits

  • Park, Sang-Yoon;Yum, Jung-Joo;Kim, Sang-Yeop
    • The Korean Journal of Zoology
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    • v.22 no.3
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    • pp.115-124
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    • 1979
  • Two homotetrameric lactate dehydrogenase isozymes from Fluta alba and one from Ophicephalus argus were purified by combination of gel filtration and DEAE-cellulose chromatogrphy. The final preparations were isozymically pure and used to elicit antibodies in rabbits. The immunochemical reactivities demonstrated that the amino acids of active site is not to be included in the antigenic determinants, that antibodies or unknown component of immunized rabbit serum might be responsible for the electrophoretic abnormality and that two subunits share common antigenic determinants, reflecting that these polypeptides have a common evolutionary origin.

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한국 진도개와 삽사리 혈액 단백질의 비교연구 II. 혈청 Lactate Dehydrogenase와 혈청 Alkaline Phosphatase의 동위효소와 활성도

  • 김종봉;윤인숙
    • The Korean Journal of Zoology
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    • v.35 no.1
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    • pp.102-106
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    • 1992
  • 진도개와 삽사리 혈청 lactate dehydrogenase와 혈청 alkaline phosphatase의 동위효소 및 효소활성도를 분석하였다. 전기영동결과 진도개와 삽사리의 혈청에서는 5두지 종류의 LDH의 동위효소가 모두 확인되었다. LDH의 활성도는 진도개의 경우 522.53 $\pm$ 279.96(U/L)이었고 삽사리는 534.10 $\pm$ 280.35(U/L)이었다. 진도개와 삽사리의 혈청 alkaline phosphatase전기 영동상에서 는 한 종류의 동위효소만 관찰되었고 활성도는 진도개의 경우 7.61 $\pm$ 4.52(K-A unit)였고 삽사리는 10.46 $\pm$ 7. 10(K-A unit) 였다. 삽사리의 ALP 활성도는 연령에 따라 커다란 차이를 나타내었다.

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Effects of Methanol Extracts of Black Soybean on Enzymes Activities of Serum in Rats Fed Pb and Cd Solution (검정콩 추출물이 납과 카드뮴을 투여한 흰쥐의 혈청 효소 활성도에 미치는 영향)

  • 한성희;신미경;김용욱;임세진
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.2
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    • pp.193-197
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    • 2000
  • The effects of black soybena extracts on enzymes activies of rat were evaluated in present study. Eighty-four male Sprague-Dawley rats weighing 100$\pm$10g were divided into twelve groups which consisted of black soybean extract, Pb and Cd solution, and black soybean extract plus Pb or Cd soln groups. The weight gain was increased in black soybean extracts and Pb soln solution group but decreased in Cd soln solution group. The results obtained form the experiment were as follows: Glutamate pyruvate trasaminase (GPT) and glutamate oxaloacetate oxaloacetate transaminase (GOT) activities were not significantly different among experimental groups. The lactate dehydrogenase (LDH) activities of black soybean extract administered groups were decreased than those of Pb and Cd solution group. Black soybean group increased cholinesterase (ChEase) activity as compared to administration of Pb and Cd soln group.

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Influences of Sodium Fluoride Contents on Hepatic Functional Enzyme Activities in Rats (Sodium Fluoride 함량이 흰쥐의 간 기능 효소활성에 미치는 영향)

  • Kim, Han-Soo
    • Journal of Environmental Science International
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    • v.28 no.11
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    • pp.943-950
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    • 2019
  • The purpose of this study was to probe the influences of NaF oral administration on a dose-effect relationship between fluoride levels of serum enzyme activity such as alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) in rats fed experimental diets for 5 weeks. All groups increased the activity of serum ALP, AST, ALT, and LDH levels with increasing NaF. In addition the fluoride levels of serum and organ tissues (liver, brain, heart, lung, kidney) in oral NaF groups (NF3~NF50) were significantly increased by adding sodium fluoride in comparison with normal diet group (ND) (p<0.05). These results, a high concentration of sodium fluoride was determined that the toxicity to various organ tissues.

Biosynthesis of Chondroitin in Engineered Corynebacterium glutamicum

  • Cheng, Fangyu;Luozhong, Sijin;Yu, Huimin;Guo, Zhigang
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.392-400
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    • 2019
  • Chondroitin, the precursor of chondroitin sulfate, which is an important polysaccharide, has drawn significant attention due to its applications in many fields. In the present study, a heterologous biosynthesis pathway of chondroitin was designed in a GRAS (generally recognized as safe) strain C. glutamicum. CgkfoC and CgkfoA genes with host codon preference were synthesized and driven by promoter Ptac, which was confirmed as a strong promoter via GFPuv reporter assessment. In a lactate dehydrogenase (ldh) deficient host, intracellular chondroitin titer increased from 0.25 to 0.88 g/l compared with that in a wild-type host. Moreover, precursor enhancement via overexpressing precursor synthesizing gene ugdA further improved chondroitin titers to 1.09 g/l. Chondroitin production reached 1.91 g/l with the engineered strain C. glutamicum ${\Delta}L-CgCAU$ in a 5-L fed-batch fermentation with a single distribution $M_w$ of 186 kDa. This work provides an alternative, safe and novel means of producing chondroitin for industrial applications.