• Title/Summary/Keyword: nucleotide-sugar

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Biochemical Characterization of a Glycosyltransferase Homolog from an Oral Pathogen Fusobacterium nucleatum as a Human Glycan-Modifying Enzyme

  • Kim, Seong-Hun;Oh, Doo-Byoung;Kwon, Oh-Suk;Jung, Jae-Kap;Lee, Yun-Mi;Ko, Ki-Sung;Ko, Jeong-Heon;Kang, Hyun-Ah
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.859-865
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    • 2008
  • Bacterial glycosyltransferases have drawn growing attention as economical enzymes for oligosaccharide synthesis, with their easy expression and relatively broad substrate specificity. Here, we characterized a glycosyltransferase homolog (Fnu_GT) from a human oral pathogen, Fusobacterium nucleatum. Bioinformatic analysis showed that Fnu_GT belongs to the glycosyltransferases family II. The recombinant Fnu_GT (rFnu_GT) expressed in Escherichia coli displayed the highest glycosylation activity when UDP-galactose (Gal) was used as a donor nucleotide-sugar with heptose or N-acetylglucosamine (GlcNAc) as an acceptor sugar. Interestingly, rFnu_GT transferred the galactose moiety of UDP-Gal to a nonreducing terminal GlcNAc attached to the trimannosyl core glycan, indicating its potential as an enzyme for human-type N-glycan synthesis.

Candidate Genes Related to Sugar Content in Sweetpotato using GWAS

  • Tae Hwa Kim;Mi Nam Chung;Hyeong Un Lee;Won Park;Sang Sik Nam
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.192-192
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    • 2022
  • Sweetpotato is rich in starch, which is converted to sugar during storage due to enzymatic hydrolysis. The sugar content of sweetpotato is a component related to taste and storability. In this study, the sugar content (fructose, glucose, maltose, sucrose and total sugar content) of 94 genotypes was evaluated and the GWAS (Genome-Wide Association Study) was conducted to search for candidate genes for sugar content. The fructose and glucose content were 0.2 ~ 8.8 and 0.2 ~ 9.4 g/100g, respectively. The maltose, sucrose and total sugar content were 0.2 ~ 9.1,3.2 - 30.0 and 7.9 ~ 40.2 g/100g, respectively. The fructose and glucose showed a positive correlation (0.98). The 94 genotypes were genotyped with genotyping-by-sequencing (GBS) and aligned against the reference genome sequences of sweetpotato. The GBS libraries from 94 genotypes were sequenced on an Illumina HiSeqXten system, and 1,339,892 SNPs (Single Nucleotide Polymorphism) were generated. Filtering for < 60% missing rate and > 0.05 minor allele frequency resulted in a total of 44,255 SNPs used in GWAS. The GAPIT (Genome Association and Prediction Integrated Tool) was used to conduct based on the mean of sugar content with a Bonferroni-corrected chromosome-wide significance threshold with a -logio(P) of 5.95. The significant SNPs were obtained with fructose (seven), glucose (six), maltose (four) and sucrose (nine). There were several genes related to sugar content around the significant SNPs such as sugar transport protein 8-like, probable galactose-1 -phosphate uridyltransferase-like and beta-amylase. These results will contribute to understanding of sugar content and conversion in sweetpotato.

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Studies on the Constituents of the Chicory Root (치코리뿌리 성분(成分)에 관(關)한 연구(硏究))

  • Kim, Taik-Young;Yoon, Young-Jin;Lee, Kyung-Woong
    • Korean Journal of Food Science and Technology
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    • v.10 no.2
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    • pp.258-262
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    • 1978
  • Proximate composition, minerals and fatty acid in dried chicory root (moisture content 7.0%) are analyzed and subsequent results are as follows: Crude protein, crude fat, crude fiber, total sugar and ash content in chicory root are 8.6%, 1.6 %, 6.9%, 58.5% and 4.2%, respectively. Mineral content of Ca, P, Fe, Mg and Si in the root are 1,560, 180, 10,600 and 180 mg%, respectively. Other minerals such as K, Na, Al, Zn, Ag, Cu and Ti are also determined. Unsaturated fatty acid content in total fat of the root is 65.4%, Particularly high in linoleic acid. Uridine-5'-diphospho-glucose, as sole nucleotide-sugar in the root, was detected.

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Chemical Components of Wid and Cultivated Horned Rampion, Phyteuma japonicum Miq. (영아자 (Phyteuma japonicum Miq)의 성분 조성)

  • 정미자;신정혜;이수정;홍성국;강호중;성낙주
    • The Korean Journal of Food And Nutrition
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    • v.11 no.4
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    • pp.437-443
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    • 1998
  • This research is to establish the basic data of the nutritive value and improve our diet. In the part of the leaf and stem of the wild and cultivated horned rampion (Phyteuma japonicum Mig), the components such as chemical compositon, vitamin C, free sugar, mineral, nucleotide and its related compounds, composition and free amino acid were analyzed one after another. Content of the crude lipids and proteins was determined much higher in its wildness than in its cultivated horned rampion; while, that of carbohydrates was higher in the former than in the latter. The content of vitamin C was retained higher in the leaf than in the stem horned rampion. And the content of calcium among the detected minerals was outstanding in all of the samples collected, and potassium and magnesium was the next ones in its order. The main components of free sugars in both the wild and cultivated horned rampion were glucose and fructose, and their content was higher in the stem than in the leaf. Nucleotide and its related compounds were identified with 5 kinds of nucieotides such as CMP, UMP, IMP, AMP and hypoxanthine (Hx), and the content of Hx and AMP was the highest I the wild and cultivated samples, respectively. In the composition amino acid of the wild horned rampion, glutamic acid, aspartic acid and phenylalanine was outstandingly abundant; while, such amino acid as methinone and proline was small and besides cysteine couldn't be detected in the stem. Total amounts of composition amino acid in the leaf was 2118.0 and 1120.1mg% in the wild and cultivated sample, respectively. In the free amino acid of horned rampion, the total amount ranged from 8.5 to 50.1mg.%, which were lower level than that of composition amino acid. But the number of free amino acid was 29 kinds, which was bigger in its number than that of composition amino acid detected 17 kinds.

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Newly Designed Six-membered Azasugar-Containing Phosphorothioate Oligonucleotide as a Potent AIDS Therapeutic Drug

  • Bae, Yong-Soo
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.155-160
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    • 2002
  • A series of modified oligonucleotides containing a phosphorothioate (P=S) backbone and a six-membered azasugar (6-AZS) as a sugar substitute in a nucleotide were synthesized and tested for their ability to inhibit the human immunodeficiency virus type I(HIV-l) in vitro without the aid of any transfecting agents. While P=S oligonucleotides with natural nucleotides had little anti-HIV-l activity, the six-membered azasugar nucleotide (6-AZN)-containing P=S oligonucleotides (AZPSONs) potently inhibited the HIV-l/SHIV replication and syncytium formation (ECso = 0.02-0.2 /lM) without cytotoxicity up to 100 /lM. DBM-2198, the most effective in anti-HIV-l activity among the AZPSONs, consists of random sequence and five 6¬AZNs evenly distributed in 18 nucleotides. DBM-2198 showed strong antiviral activity against, not only laboratory strains, but also primary isolates and even drug-resistant strains of HIV-I. DBM-2198 was much more effective than ddI or ddC in its anti-HIV-l activity in vitro. Particularly noteworthy is that the anti-HIV-l activity of DBM-2198 was better than that of AZT with respect to its long-lasting efficacy after a single treatment. Nevertheless, the antiviral activity of the AZPSONs was very specific to HIV-I. Poliovirus, or even simian immunodeficiency virus (SIV), was not inhibited by the AZPSONs. Taken together, our results strongly suggest that AZPSON can be used as a safe and effective AIDS-therapeutic drug against a broad spectrum of HIV -1 strains.

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Diagnosis of Salmonella dubin in Korean Native Calves using PCR and Nucleotide Sequences of rfb5 Gene (송아지에 감염된 Salmonella dublin의 PCR 진단과 rfbS 항원단백 유전자의 염기서열분석)

  • 김철민;이영준;박명규;최경성;김민석
    • Journal of Veterinary Clinics
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    • v.17 no.2
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    • pp.464-469
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    • 2000
  • An epizootic of calf diarrhea occurred in a Korean native cattle farm located in Chonbuk province. The calves that had either bloody or watery diarrhea were 1 to 30 days old. Some of these animals died during the acute course of the disease. Five calves with predominant clinical signs were examined in more detail. Hematological and serum chemical findings were suggestive of dehydration and nutritional insufficiency. Fecal material from the calve was cultured on/in brilliant green agar (BGA), xylose-lysine deoxycholate (XLD) medium, MacConkey agar, eosin methylene blue (EMB) agar and triple sugar iorn (TSI) A bacteria was isolated. which was subsequently identificed as belonging to Salmonella spp. To differentiate Salmoenlla serotype, rfbs gene of S. dublin was amli- find (720 bp) by multiplex (PCR). The rfbS gene sequences of S, dublin ficld isolate(SDC-1) was com- pared with that off S. dublin(S-37) S, dublin(Ahn et al, 1996), S enteritidis(Ahn et al 1996)and S. typhi (Generbak accession No M29682). The identities of nucleotide sequences were 100%. 99.6%, 99.6%, 97.5% respectively.

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Nucleotide Sequence and Cloning of sfs4, One of the Genes Involved in the CRP-Dependent Expression of E. coli mal Genes. (CRP 의존성 maltose 대사 촉진 유전자 sfs4의 클로닝 및 염기배열 결정)

  • Chung, Soo-Yeol;Cho, Moo-Je;Jeong, Hee-Tae;Choi, Yong-Lark
    • Applied Biological Chemistry
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    • v.38 no.2
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    • pp.111-117
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    • 1995
  • In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

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Biochemical Characterization of a Putative Calcium Influx Factor as a Diffusible Messenger in Jurkat Cells, Xenopis Oocytes, and Yeast

  • Kim, Hak-Yong
    • Animal cells and systems
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    • v.7 no.1
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    • pp.75-79
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    • 2003
  • Highly purified high performance thin layer chromatography (HPTLC) fractions containing a putative calcium influx factor (CIF) were prepared from the Jurkat cells and Xenopus oocytes in which $Ca^{2+}$ stores were depleted by thapsigargin treatment and from the yeast in which intracellular $Ca^{2+}$ stores were also depleted by genetic means. Microinjection of the fractions has been shown to elicit $Ca^{2+}$ dependent currents in Xenopus oocytes. The nature of the membrane currents evoked by the putative CIF appeared to be carried by chloride ions since the current was blocked by the selective chloride channel blocker 1 mM niflumic acid and its reversal potential was about -24 mV. Injection of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid (BAPTA) eradicated the current activities, suggesting the current responses are entirely $Ca^2$-dependent. Moreover, the currents were sensitive to the removal of extracellular calcium, indicating the dependence on calcium entry through the plasma membrane calcium entry channels. CIF activities were insensitive to protease, heat, and acid treatments and to Dische-reaction whereas the activities were sensitive to nucleotide pyrophosphatase and hydrazynolysis. The fraction might have a sugar because it was sensitive to Molisch test and Seliwaniff's resorcinol reaction. From the above results, CIF as a small and stable molecule seems to have pyrimidine, pyrophosphate, and a sugar moiety.oiety.