• Title/Summary/Keyword: de novo synthesis

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Differential Induction of Pathogenesis-Related Proteins in the Compatible and Incompatible Interactions of Tomato Leaves with Xanthomonas campestris pv. vesicatoria (Xanthomonas campestris pv. vesicatoria와 토마토잎의 친화적, 불친화적 반응에서 병생성관련 단백질의 유도)

  • 김정동;황병국
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.53-60
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    • 1995
  • Inoculation with the compatible strain Ds 1 of Xanthomonas campestris pv. vesicatoria caused brownish ad water-soaked lesions, but incompatible strain Bv5-4a produced hypersensitive symptoms with local necrosis on tomato (cv. Kwangyang) leaves. Bacterial populations of the compatible strains Ds 1 propagated more greatly than the incompatible strain Bv5-4a at the frist onset, but no differences were observed 5 days after inoculation. The bacterial infection induced the synthesis and accumulation of soluble proteins in tomato leaves, especially in the incompatible interaction. Native-polyacrylamide gel electrophoresis distinguished the soluble proteins in the tomato leaves infected by the compatible or incompatible strains. A protein of low molecular weight occurred only in the incompatible interaction. Some pathogenesis-related (PR) proteins, especially the 15, 18, 23, 26 and 54 kDa proteins, were detected only in the infected tomato leaves. In the two-dimensional electrophoresis, some proteins with different molecular weights (Mr. 21∼29 kDa) and the pI 8∼9 appeared more distinctly only in the incompatible interaction. These data suggest that the de novo synthesis of some PR proteins in tomato may be significant in defense against X. c. pv. vesicatoria.

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Utilization of Putrescine by Streptococcus pneumoniae During Growth in Choline-limited Medium

  • Ware D.;Watt J.;Swiatlo E.
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.398-405
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    • 2005
  • Polyamines such as putrescine are small, ubiquitous polycationic molecules that are required for optimal growth of eukaryotic and prokaryotic cells. These molecules have diverse effects on cell physiology and their intracellular content is regulated by de novo synthesis and uptake from the environment. The studies presented here examined the structure of a putative polyamine transporter (Pot) operon in Streptococcus pneumoniae (pneumococcus) and growth of pneumococci in medium containing putrescine substituted for choline. RT-PCR experiments demonstrated that the four genes encoding the Pot system are co-transcribed with murB, a gene involved in an intermediary step of peptidoglycan synthesis. Pneumococci grown in chemically-defined media (CDM) containing putrescine without choline enter logarithmic phase growth after 36-48 hs. However, culture density at stationary phase eventually reaches that of choline-containing medium. Cells grown in CDM-putrescine formed abnormally elongated chains in which the daughter cells failed to separate and the choline-binding protein PspA was no longer cell-associated. Experiments with CDM containing radiolabeled putrescine demonstrated that pneumococci concentrate this polyamine in cell walls. These data suggest that pneumococci can replicate without choline if putrescine is available and this polyamine may substitute for aminoalcohols in the cell wall teichoic acids.

Triclosan Inhibition of Prostaglandin $E_2$ Production in Human Gingival Fibroblast (치은 섬유모세포에서 Triclosan에 의한 Prostaglandin $E_2$ 합성 억제)

  • Park, Seong-Pyu;Chung, Hyun-Ju;Kim, Young-Joon;Kim, Ok-Su
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.345-356
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    • 2004
  • The triclosan was shown to have anti-microbial and anti-inflammatory effect with inhibition of inflammatory mediators such as prostaglandin $E_2(PGE_2)$. The purpose of this study was to elucidate whether and how $PGE_2$ could be inhibited by triclosan in human gingival fibroblast. Human gingival fibroblast-1 cells (ATCC CRL2014) were pre-treated for 1 hour with triclosan (0.001 ${\mu}/ml{\sim}10$ ${\mu}/ml$) and then stimulated with $TNF-{\alpha}$ (1.0 ng/ml). $PGE_2$ synthesis was evaluated by ELISA and gene expression of COX-1 and COX-2 was evaluated by RT-PCR after $TNF-{\alpha}$, triclosan, and NS-398 (COX-2 inhibitor, 5, ${\mu}M$) and/ or cycloheximide (protein synthesis inhibitor, 2 ${\mu}g/ml$). Triclosan was cytotoxic to human gingival fibroblasts in the concentration higher than 1.0 ${\mu}g/ml$ for longer than 24 hours in tissue culture. The $PGE_2$ synthesis was inhibited by triclosan in dose-dependent manner. Greater COX-2 mRNA suppression was observed with triclosan (0.1 ${\mu}g/ml$) than with $TNF-{\alpha}$ alone, without change in COX-1 gene expression. Inhibitory effects of triclosan on $PGE_2$ synthesis disappeared in presence of cycloheximide. This study suggests that triclosan inhibit prostaglandin $E_2$ at the level of COX-2 gene regulation and require de novo protein synthesis.

Alpha-Lipoic Acid Inhibits Glycogen Synthesis and Modifies Glucose Metabolism and Signaling Pathways in Soleus Muscles from Healthy Rats

  • Madar, Zecharia;Stark, Aliza H.;Ilan, Erez;Timar, Bracha;Borenshtein, Diana
    • Preventive Nutrition and Food Science
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    • v.7 no.2
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    • pp.113-118
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    • 2002
  • Alpha-lipoic acid is a known hypoglycemic agent that may be useful in the treatment of diabetes. The objective of this study was to investigate the fate of glucose in isolated muscles incubated with lipoic acid by determining its direct effects on specific metabolic and signaling pathways. Soleus muscles from healthy rats were incubated with lipoic acid in the absence or presence of insulin. Glucose transport, glycogen synthesis, glucose oxidation and lipid synthesis were determined and affects on major pathways associated with insulin signaling were evaluated. Glucose transport was not significantly altered by the addition of lipoic acid to the incubation medium. However, lipoic acid decreased glycogen synthesis in comparison to controls. Glucose oxidation was moderately increased while de-novo lipid synthesis from glucose was inhibited. Wortmannin repressed insulin stimulation of glucose incorporation into glycogen, an effect that was augmented by the combined treatment of wortmannin and lipoic acid. Basal and insulin-stimulated serine phosphorylation of Akt was not changed by the addition of lipoic acid to the incubation medium. These data show that in this in vitro model, lipoic acid did not significantly affect glucose uptake but dramatically modified pathways of glucose metabolism within muscle tissue.

Supplementation of Either Conjugated Linoleic Acid or γ-linolenic Acid with or without Carnitine to Pig Diet Affect Flavor of Pork and Neutrophil Phagocytosis

  • Lee, Jun-Yeob;Cha, Keun-Hwan;Chae, Byong-Jo;Ohh, Sang-Jip
    • Journal of Animal Science and Technology
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    • v.53 no.3
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    • pp.237-252
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    • 2011
  • In this study, four different oils containing either CLA, GLA, GLA+Carnitine or corn oil (control) were supplemented to finishing pigs (average 70.8 kg initial BW) diet for 28 d of feeding period. To evaluate the values of the dietary fatty acids, especially in view of sensory and nutritional characteristics of pork; pig performances, carcass characteristics, serum cholesterol, neutrophil phagocytosis, TBARS, electronic nose flavor and fatty acids profile of pork were measured. There were no differences in daily gain and nutrients digestion among treatments, but daily feed intake of CLA enriched diet was lower (P<0.05) than that of other diets. There were no differences in backfat thickness, dressing percentage and carcass grade among pigs fed diets supplemented with different oils. Serum total cholesterol showed a tendency to be lowered in pigs fed GLA enriched diet. TBARS values during storage of pork were higher in belly from pigs fed control diet whereas the values of belly from pigs fed GLA+Carnitine diet were lower than others. However, difference in TBARS was not remarkable in adipose tissue and 4 weeks extended storage regardless of pork parts. Proportion of saturated fatty acids such as C16:0 and C18:0 were higher (P<0.05) in pork loin and thin skirt from pigs fed CLA enriched diet compared to those from other diets. There were no differences in fatty acids profiles of belly and adipose tissue. CLA accumulation in pork was increased by the dietary CLA supplementation and this could be also confirmed by a slight de novo synthesis of CLA in pork from pigs fed CLA free diets. GLA was selectively accumulated to pork adipose tissue and loin from pigs fed GLA enriched diets. There was no accumulation of GLA when GLA was not supplemented, indicating no de novo synthesis of GLA. Phagocytic activity was the highest (p<0.05) in neutrophil of pigs fed GLA+Carnitine supplemented diet, then, followed by pigs fed GLA supplemented diet. There was no difference in phagocytosis between control and CLA treatment although the phagocytosis was numerically lowest in pig fed CLA enriched diet. There were distinct differences in electronic nose flavor pattern among treatments regardless of the parts. This study showed that dietary supplementation of functional fatty acids like CLA or GLA was able to result in characteristic differences in feed intake, TBARS, fatty acids profile and flavor of pork, serum cholesterol regulation and neutrophil phagocytosis.

Beneficial Effect of Collagen Peptide Supplement on Anti-aging Against Photodamage (콜라겐 펩타이드의 피부 광노화 예방 효과)

  • Kim, Jeong-Kee;Lee, Ji-Hae;Yang, Mi-Sook;Seo, Dae-Bang;Lee, Sang-Jun
    • Korean Journal of Food Science and Technology
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    • v.41 no.4
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    • pp.441-445
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    • 2009
  • Recent research has revealed that hydrolyzed collagen peptides have beneficial effects in various diseases such as osteoarthritis and human rheumatoid arthritis and also play a protective role in skin by improving the activity of antioxidants. In this study, we investigated the effects of a novel mixture (AP-CPM01) containing collagen peptides and elastin peptides on photoaged hairless mice skin both in vivo and in vitro. To evaluate the effects of AP-CPM01 on UVBinduced skin wrinkle formation in vivo, the hairless mice were exposed to UVB irradiation and orally administered the AP-CPM01 at 333 mg/kg per day for 10 weeks. The effects on skin appearance and epidermal thickness were measured using bioengineering and histochemical methods. In addition, the influence of AP-CPM01 on collagen metabolism in human skin fibroblasts was also investigated. The skin of mice in the AP-CPM01 treated group had better appearance and less wrinkling than that of mice in the control group. In the human fibroblast cells, the amount of de novo procollagen synthesis was increased after AP-CPM01 treatment, reflecting that AP-CPM01 can induce de novo procollagen synthesis and reduce UVB-induced skin wrinkle formation. These results suggest that AP-CPM01 is a potent candidate for antiphotoaging functions.

Biosynthesis of the extracellular enzymes in de novo during the differentiation of Aspergillus niger (검정곰팡이의 형태분화에 따른 세포외성효소의 신생적생합성에 관한 연구)

  • Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.6 no.2
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    • pp.1-10
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    • 1978
  • In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.

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Effects of pyrimidine salvage inhibitors on uracil incorporation of Toxoplasma gondii (Toxoplasma gondii의 활성화된 uracil 도입 과정에 미치는 pyrimidine 대사 억제제의 영향)

  • 윤지혜;남호우
    • Parasites, Hosts and Diseases
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    • v.28 no.2
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    • pp.79-84
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    • 1990
  • Metabolic inhibitors which act in the process of pyrimidine salvage influenced on the uracil incorporation into nucleic acids of Toxoplasma. Inhibitors of dihydrofolate reductase, pyrimethamine and methotrexate, and inhibitors of thymidylate synthase, fluoro-uridine, fluoro·dUMP and fluoro-uracil, diminished isotopic uracil uptake in dose-dependent manners. Azauridine which suppresses do novo pyrimidine biosynthesis did not affect the salvage even in a relatively high dose. These results suggested that the activation of uracil salvage should be closely related with the function of TMP biosynthetic enzymes. The pattern of thymidine uptake had no differences between control HL-60 cells and Toxoplasma infected cells, which did not reject the specific proliferation of Texoplasma. It can be exploited to characterize the elects of various compounds related with the proliferation of Toxoplasma, especially its DNA synthesis. Key words: Toxoplasma gondii, uracil salvage, dihydrofolate reductase, thymidylate synthase TMP biosynthesis.

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Sources and Variations of Extracellular Enzymes in a Wetland Soil (습지 토양에서 체외효소의 근원과 변화)

  • Freeman, Chris;Kang, Ho-Jeong
    • Korean Journal of Ecology and Environment
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    • v.35 no.4 s.100
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    • pp.326-330
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    • 2002
  • A wetland soil was sterilised by two methods and changes in microbial enzyme activities were assessed. The short-term effects were determined by toluene addition, while the longer-term effects of elimination was monitored by ${\gamma}$-radiation. The changes in ${\beta}$- glucosidase, ${\beta}$-xylosidase, cellobiohydrolase, phosphatase, arylsulphatase, and N-acetylglucosaminidase activities were determined by using methylumbelliferyl model substrates and comparing with the activities of control samples. Toluene addition induced different responses of enzymes. For example, phosphatase activity increased by the treatment while ${\beta}$-glucosidase and arylsulphatase activities decreased. In contrast, ${\gamma}$-radiation decreased all enzyme activities compared to control by 40-80%. The overall results of the toluene and ${\gamma}$-radiation experiments indicate that the large amounts of enzymes are stabilised outside of living cells, at least in the short term, but that the persistence of enzymes is maintained by de-novo synthesis of microbes.

Impact of High-Level Expression of Heterologous Protein on Lactococcus lactis Host

  • Kim, Mina;Jin, Yerin;An, Hyun-Joo;Kim, Jaehan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1345-1358
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    • 2017
  • The impact of overproduction of a heterologous protein on the metabolic system of host Lactococcus lactis was investigated. The protein expression profiles of L. lactis IL1403 containing two near-identical plasmids that expressed high- and low-level of the green fluorescent protein (GFP) were examined via shotgun proteomics. Analysis of the two strains via high-throughput LC-MS/MS proteomics identified the expression of 294 proteins. The relative amount of each protein in the proteome of both strains was determined by label-free quantification using the spectral counting method. Although expression level of most proteins were similar, several significant alterations in metabolic network were identified in the high GFP-producing strain. These changes include alterations in the pyruvate fermentation pathway, oxidative pentose phosphate pathway, and de novo synthesis pathway for pyrimidine RNA. Expression of enzymes for the synthesis of dTDP-rhamnose and N-acetylglucosamine from glucose was suppressed in the high GFP strain. In addition, enzymes involved in the amino acid synthesis or interconversion pathway were downregulated. The most noticeable changes in the high GFP-producing strain were a 3.4-fold increase in the expression of stress response and chaperone proteins and increase of caseinolytic peptidase family proteins. Characterization of these host expression changes witnessed during overexpression of GFP was might suggested the metabolic requirements and networks that may limit protein expression, and will aid in the future development of lactococcal hosts to produce more heterologous protein.