• Title/Summary/Keyword: aldolase

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Synthesis of L-threo-2,3-Dihydroxyphenylserine (L-threo-DOPS) by Thermostable L-Threonine Aldolase Expressed in Corynebacterium glutamicum R (Corynebacterium glutamicum에서 발현된 L-Threonine Aldolase를 이용한 파킨슨병 치료제 L-threo-2,3-Dihydroxyphenylserine (L-threo-DOPS)의 합성)

  • Baik, Sang-Ho
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.128-134
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    • 2008
  • In order to examine efficient L-threo-2,3-Dihydroxyphenylserine (L-threo-DOPS) synthesis process using whole cell biocatalyst, a thermostable L-threonine aldolase (L-TA), which cloned from Streptomyces coelicolor A3(2) and improved for stability, was expressed in a Corynebacterium glutamicum R strain. The constructed Corynebacterium expression vector, pCG-H44(1) successfully expressed L-TA in C. glutamicum R strain, but showed very low expression level. In order to improve the expression level, the expression vector named pCG-H44(2) was reconstructed by eliminating 1 nucleotide between SD sequence and start codon of L-TA. The pCG-H44(2) vector plasmid was able to overexpress L-TA approximately 3.2 times higher than pCG-H44(1) in C. glutamicum R strain (CGH-2). When the whole cell of CGH-2 was examined in a repeated batch system, L-threo-DOPS was successfully synthesized with a yield of 4.0 mg/ml and maintain synthesis rate constantly after 30 repeated batch reactions for 130 h.

Molecular Cloning and Characterization of Salt-inducible Aldolase from Salicornia herbacea (퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석)

  • Cha, Joon-Yung;Netty Ermawati;Kim, Soon-Gil;Lee, Jeung-Joo;Lim, Chae-Oh;Chung, Woo-Sik;Lee, Kon-Ho;Son, Dae-Young
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.323-328
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    • 2003
  • Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

Effects of Cibotii Rhizoma(狗脊) on the Muscle Atrophy Induced by Hindlimb Suspension in Rats (구척(狗脊)이 흰쥐의 후지체공(後肢滯空)에 의한 근강축(筋萎縮)에 미치는 영향(影響))

  • Kim, Yeong-Gyun;Kim, Jong-Hun
    • The Journal of Internal Korean Medicine
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    • v.17 no.1
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    • pp.51-65
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    • 1996
  • This experimental study was designed to investigate the effects of Cibotii Rhizoma on the muscle atrophy induced by hindlimb suspension in rats. The measurement has been performed on the activity of CK, aldolase, LDH, AST, ALT and quantity of creatine in serum of hindlimb suspension rats. The results were as ; 1. Cibotii Rhizoma significantly inhibited the increase of the activity of CK in serum. 2. Cibotii Rhizoma significantly inhibited the increase of the quantity of creatine in serum. 3. Cibotii Rhizoma significantly inhibited the increase of the activity of aldolase in serum. 4. Cibotii Rhizoma significantly inhibited the increase of the activity of LDH in serum. 5. Cibotii Rhizoma significantly inhibited the increase of the activity of AST in serum. 6. Cibotii Rhizoma significantly inhibited the increase of the activity of ALT in serum.

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Crystallographic snapshots of active site metal shift in E. coli fructose 1,6-bisphosphate aldolase

  • Tran, Huyen-Thi;Lee, Seon-Hwa;Ho, Thien-Hoang;Hong, Seung-Hye;Huynh, Kim-Hung;Ahn, Yeh-Jin;Oh, Deok-Kun;Kang, Lin-Woo
    • BMB Reports
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    • v.49 no.12
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    • pp.681-686
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    • 2016
  • Fructose 1,6-bisphosphate aldolase (FBA) is important for both glycolysis and gluconeogenesis in life. Class II (zinc dependent) FBA is an attractive target for the development of antibiotics against protozoa, bacteria, and fungi, and is also widely used to produce various high-value stereoisomers in the chemical and pharmaceutical industry. In this study, the crystal structures of class II Escherichia coli FBA (EcFBA) were determined from four different crystals, with resolutions between $1.8{\AA}$ and $2.0{\AA}$. Native EcFBA structures showed two separate sites of Zn1 (interior position) and Zn2 (active site surface position) for $Zn^{2+}$ ion. Citrate and TRIS bound EcFBA structures showed $Zn^{2+}$ position exclusively at Zn2. Crystallographic snapshots of EcFBA structures with and without ligand binding proposed the rationale of metal shift at the active site, which might be a hidden mechanism to keep the trace metal cofactor $Zn^{2+}$ within EcFBA without losing it.

Expression and Characterization of a Novel Deoxyribose 5-Phosphate Aldolase from Paenibacillus sp. EA001

  • Kim, Yong-Mo;Choi, Nack-Shick;Kim, Yong-Ook;Son, Dong-Ho;Chang, Young-Hyo;Song, Jae-Jun;Kim, Joong-Su
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.995-1000
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    • 2010
  • A novel deoC gene was identified from Paenibacillus sp. EA001 isolated from soil. The gene had an open reading frame (ORF) of 663 base pairs encoding a protein of 220 amino acids with a molecular mass of 24.5 kDa. The amino acid sequence was 79% identical to that of deoxyribose 5-phosphate aldolase (DERA) from Geobacillus sp. Y412MC10. The deoC gene encoding DERA was cloned into an expression vector and the protein was expressed in Escherichia coli. The recombinant DERA was purified using Ni-NTA affinity chromatography and then characterized. The optimum temperature and pH of the enzyme were $50^{\circ}C$ and 6.0, respectively. The specific activity for the substrate deoxyribose 5-phosphate (DR5P) was $62\;{\mu}mol/min/mg$. The $K_m$ value for DR5P was determined to be 145 mM with the $k_{cat}$ value of $3.2{\times}10^2/s$ from Lineweaver-Burk plots. The EA001 DERA showed stability toward a high concentration of acetaldehyde (100 mM).

Protective and Anti-Pathology Effects of Sm Fructose-1,6-Bisphosphate Aldolase-Based DNA Vaccine against Schistosoma mansoni by Changing Route of Injection

  • Saber, Mohamed;Diab, Tarek;Hammam, Olft;Karim, Amr;Medhat, Amina;Khela, Mamdouh;El-Dabaa, Ehab
    • Parasites, Hosts and Diseases
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    • v.51 no.2
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    • pp.155-163
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    • 2013
  • This study aimed to evaluate the efficacy of fructose-1,6-bis phosphate aldolase (SMALDO) DNA vaccination against Schistosoma mansoni infection using different routes of injection. The SMALDO has been cloned into the eukaryotic expression vector pcDNA3.1/V5-His TOPO-TA and was used in injecting Swiss albino mice intramuscularly (IM), subcutaneously (SC), or intraperitoneally (IP) ($50{\mu}g/mouse$). Mice vaccinated with non-recombinant pcDNA3.1 served as controls. Each group was immunized 4 times at weeks 0, 2, 4, and 6. Two weeks after the last booster dose, all mice groups were infected with 80 S. mansoni cercariae via tail immersion. At week 8 post-infection, animals were sacrificed for assessment of parasitological and histopathological parameters. High anti-SMALDO IgG antibody titers were detected in sera of all vaccinated groups (P<0.01) compared to the control group. Both the IP and SC vaccination routes resulted in a significant reduction in worm burden (46.2% and 28.9%, respectively, P<0.01). This was accompanied by a significant reduction in hepatic and intestinal egg counts (41.7% and 40.2%, respectively, P<0.01) in the IP group only. The number of dead eggs was significantly increased in both IP and IM groups (P<0.01). IP vaccination recorded the highest significant reduction in granuloma number and diameter (54.7% and 29.2%, respectively, P<0.01) and significant increase in dead miracidia (P<0.01). In conclusion, changing the injection route of SMALDO DNA vaccination significantly influenced the efficacy of vaccination. SMALDO DNA vaccination via IP route could be a promising protective and antipathology vaccine candidate against S. mansoni infection.

Glyceraldehyde-3-Phosphate, a Glycolytic Intermediate, Plays a Key Role in Controlling Cell Fate Via Inhibition of Caspase Activity

  • Jang, Mi;Kang, Hyo Jin;Lee, Sun Young;Chung, Sang J.;Kang, Sunghyun;Chi, Seung Wook;Cho, Sayeon;Lee, Sang Chul;Lee, Chong-Kil;Park, Byoung Chul;Bae, Kwang-Hee;Park, Sung Goo
    • Molecules and Cells
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    • v.28 no.6
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    • pp.559-563
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    • 2009
  • Glyceraldehyde-3-phosphate is a key intermediate in several central metabolic pathways of all organisms. Aldolase and glyceraldehyde-3-phosphate dehydrogenase are involved in the production or elimination of glyceraldehyde-3-phosphate during glycolysis or gluconeogenesis, and are differentially expressed under various physiological conditions, including cancer, hypoxia, and apoptosis. In this study, we examine the effects of glyceraldehyde-3-phosphate on cell survival and apoptosis. Overexpression of aldolase protected cells against apoptosis, and addition of glyceraldehyde-3-phosphate to cells delayed apoptosis. Additionally, delayed apoptotic phenomena were observed when glyceraldehyde-3-phosphate was added to a cell-free system, in which artificial apoptotic process was induced by adding dATP and cytochrome c. Surprisingly, glyceraldehyde-3-phosphate directly suppressed caspase-3 activity in a reversible noncompetitive mode, preventing caspase-dependent proteolysis. Based on these results, we suggest that glyceraldehyde-3-phosphate, a key molecule in several central metabolic pathways, functions as a molecule switch between cell survival and apoptosis.

Effect of Honey on the Change of Enzyme Activity in Rats (벌꿀이 흰쥐의 효소활성에 미치는 영향)

  • 정동현;백승화;박성수
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.65-73
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    • 1997
  • The purpose of the study was to find an effect of honey on the enzyme activity of Sprague Dawley rats. All experimental rats were fed ad libitum for seven weeks with 68% saccharide diet and at same time fed administratively with 10% and 20% water solution of acacia, sumac, polyflower honey, and sucrose, respectively. The level of LDH activity in serum of rat taken diet with acasia, sumac, and polyflower honey were increased in comparison with the control group. The level of $\alpha$-HBDH activity in serum of rat taken diet with acasia, sumac, polyflower honey, and sugar solution were increased than that other honey solution. The level of GOT and GPT activity in serum was increased by the feeding of solution of 20% acacia honey. The level of ICD activity in serum of rat taken diet with sumac honey was increased but was decreased notably by the feeding of polyflower honey. The level of G-6-P DH activity in whole blood of rat taken diet with honey solutions were decreased, but the level of aldolase activity in serum of rat taken diet with honey solutions were increased.

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Production of glycerol from glucose by dunaliella tertiolecta cell-free systems (Dunaliella tertiolecta cell-free system에 의한 글리세롤의 성장)

  • 권영명
    • Korean Journal of Microbiology
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    • v.8 no.1
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    • pp.35-40
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    • 1970
  • In the cell-free systems of Dunaliella tertiolecta, fructosediphosphate aldolase hardly contribute to synthesize hexosephosphate from triosephosphate derived from pentosphosphate pathway, and it could be considered that glycerol synthesized from added glucose was synthesized but via 3-phosphoglyceraldehyde as an intermediate not hydroxypyruvate.

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Glycosylation of Protein by Conjugation of Periodate-Oxidized Sugars (과요오드산 산화당에 의한 인공 당단백질의 조제)

  • Ann, Yong-Geun
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.62-67
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    • 1999
  • Periodate-oxidized soluble starch and maltohexaose, maltotetraose, maltose, and glyceraldehyde reacted with sweet potato ${\beta}-amylase$, wheat ${\beta}-amylase$, aldolase, bovine serum albumin, catalase, carboxypeptidase, ferritin and pronase. Electrophoretical mobility of modified proteins was different from that of native proteins, and modified proteins were stained with periodic acid-Schiff while native proteins did not stain. This results means that oxidized sugars attached on proteins. This bond is based on the Schiffs base between CHO group of oxidized sugar and ${\varepsilon}-NH_2$ group of lysine of protein. There is no changed UV absorption spectrum of sweet potato ${\beta}-amylase$ modified with oxidized soluble starch, in comparison with native enzyme.

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