• Title/Summary/Keyword: lysA

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PHOTOCATALYTIC SYNTHESIS OF L-PIPECOLINIC ACID FROM $N_{varepsilon}$-CARBAMYL-L-LYSINE BY AQUEOUS SUSPENSION OF PLATINIZED TITANIUM(IV) OXIDE

  • Ohtani, Bunsho;Aoki, Eishiro;Iwai, Kunihiro;Nishimoto, Sei-Ichi
    • Journal of Photoscience
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    • v.1 no.1
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    • pp.31-37
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    • 1994
  • Photoirradiation at > 300 nm onto a suspension of platinized TiO$_2$ (TiO$_2$-Pt) particles in an aqueous solution. of N$_{\varepsilon}$-carbamyI-L-lysine (Lys(CONH)$_2$) induced the selective N-cyclization of Lys(CONH$_2$) into almost optically pure L-pipecolinic acid (PCA) under argon atmosphere at ambient temperature. Among various TiO$_2$-Pt catalysts, a P-25 (Degussa) powder platinized via impregnation from chloroplatinic acid followed by hydrogen reduction at 753 K exhibited the highest photocatalytic activity for Lys(CONH$_2$) consumption and L-PCA production. GC-MS analyses of L-PCA obtained photocatalytically from $^{15}$N$\alpha$-Lys(CONH$_2$) revealed the selective formation $^{15}$N-substituted L-PCA. This implies that the mechanism for L-PCA production contains selective cleavage of C$_{\varepsilon}$-N bond and intramolecular alkylation at $\alpha$-amino group. Effect of pH on the rate of this photocatalytic reaction was investigated in detail and compared with the pH-dependent charge distribution in Lys(CONH$_2$) molecule. It is clarified that protonation-deprotonation of $\alpha$-amino group gives marked influence on the rate and selectivity of the photocatalytic reaction. On the basis of these results, it is concluded that the selective production of optically pure L-PCA, especially in an acidic suspension of TiO$_2$-Pt, was attributed to the enhanced protonation of $\alpha$-amino group to prevent undesirable oxidation by photogenerated positive holes and blocking of $\varepsilon$-amino group to yield racemic Schiff base intermediate.

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Evaluation of antibacterial and therapeutic effects of egg-white lysozyme against Salmonella Typhimurium in ICR mice infected with Salmonella Typhimurium (난백 유래 라이소자임의 마우스 살모넬라증에 대한 항균 및 치료 효과)

  • Kim, Hee-Gyu;Hwang, Jae-Seung;Jae, Woo-Young;Son, Song-Ee;Lee, Hu-Jang
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.103-108
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    • 2016
  • Salmonellosis is a major bacterial zoonosis that causes self-limited enteritis in animals and foodborne disease and typhoid fever in humans. Recently, multi-drug-resistant strains of Salmonella spp. have increased and caused more serious problems in public health. The present study investigated the antibacterial effects of egg-white lysozyme (EWL) against Salmonella (S.) Typhimurium and the therapeutic effects of EWL for murine salmonellosis. Evaluation of the antibacterial effects of EWL against S. Typhimurium revealed a minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EWL of 6.25 and $300{\mu}g/mL$, respectively. In the bacterial growth inhibition test, EWL at 300 (p < 0.05) and $600{\mu}g/mL$ (p < 0.01) significantly inhibited the growth of S. Typhimurium at 4 h post-incubation. EWL administration at MIC (LYS-1), MBC (LYS-2) and $2{\times}MBC$ (LYS-3) for 14 days resulted in mortality of mice infected with S. Typhimurium of 70, 40 and 10%, respectively, while that of control mice (CON) was 90%. Counts of S. Typhimurium in murine spleens were significantly lower in LYS-2 and LYS-3 than CON (p < 0.05). The results of this study indicate that EWL has the potential for treatment of ICR mice infected with S. Typhimurium.

Regulatory Mechanism of Lysine Biosynthetic Genes in Escherichia coli

  • Joe, Min-Ho;Mun, Hyo-Young;Hong, Mi-Ju;Kim, Seong-Jun;Park, Young-Hoon;Rhee, Sang-Ki;Kwon, Oh-Suk
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.236-241
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    • 2004
  • In Escherichia coli, L-lysine biosynthetic pathway is composed of nine enzymatic reactions. It has been well established that most of the lysine biosynthetic genes are regulated by the lysine availability, even though they are all scattered around the chromosome without forming any multigenic operon structure. However, no transcriptional regulatory mechanism has been identified except for the activation of lysA gene by the LysR. In this study, changes in transcriptome profiles of wild type cells and lysR deletion mutant cells grown in the absence or presence of lysine were investigated by using DNA microarray technique. Microarray data analysis revealed three groups of genes whose expression varies depending on the availability of lysine or LysR or both. To further examine the regulatory patterns of lysine biosynthetic genes, lacZ operon fusions were constructed and their expression was measured under various conditions. Obtained results strongly suggest that there is an additional regulatory mechanism which senses the lysine availability and coordinates gene expression.

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Thymol and eugenol in essential oils enhance phage endolysin LysECP26-mediated cell wall disruption of Escherichia coli O157:H7

  • Park, Do-Won;Lee, Jong Hun;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.819-822
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    • 2021
  • To enhance phage endolysin-mediated cell wall disruption of Escherichia coli O157:H7, the cells were co-treated with aromatic compounds, namely thymol or eugenol, found in essential oils and endolysin LysECP26. Interestingly, the minimal inhibitory concentrations of LysECP26 was four times lower when used in combination with either of the two compounds than when it was used alone. This synergistic activity was also confirmed by viable cell counting. Within 1 h of LysECP26 and eugenol or thymol co-treatment to the cells, there was a 2.3 or 3.8 log CFU/mL reductions, respectively. Additionally, field emission scanning electron microscopy showed cell wall disruption and severe morphological alterations of the cells in case of the combination treatments. Therefore, endolysin and thymol or eugenol co-treatment can help in developing efficient bio-control strategies against gram-negative pathogen E. coli O157:H7.

Development of Substrate for Carboxypeptidase-B by Employing Thiaarginine Peptides

  • 홍남주;박영애;손기남
    • Bulletin of the Korean Chemical Society
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    • v.19 no.2
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    • pp.189-193
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    • 1998
  • Carboxypeptidase-B (CPB) is involved in the biosynthesis of numerous peptide hormones and neurotransmitters. CPB catalyzes hydrolysis of the basic amino acids from the C-terminal position in polypeptides during posttranslational prohormonal processing. Various peptides containing thiaarginine residue at C-terminal position were synthesized and tested for their hydrolysis by CPB. A colorimetric assay, employing Ellman's reagent to detect the thioguanidine released upon hydrolysis of the dipeptide substrates, showed that thiaarginine is a suitable mimetic for arginine. Kinetic studies on the four substrates, Z-L-Ala-DL-thia-Lys, Z-L-Ala-DL-thia-Arg, Z-L-Lys-DL-thia-Arg, and Z-L-Lys(Boc)-DL-thia-Arg, gave Km (mM) of 0.66, 5.08, 0.024, and 0.006 and kcat (min-1) of 340, 5200, 151 and 335, respectively.

Roles of the Residues Lys115 and Tyr116 in the Binding of an Allosteric Inhibitor AMP to Pea Cytosolic Fructose-1,6-bisphosphatase

  • Jang, Hye-Kyung;Cho, Man-Ho;Kwon, Yong-Kook;Bhoo, Seong-Hee;Jeon, Jong-Seong;Hahn, Tae-Ryong
    • Journal of Applied Biological Chemistry
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    • v.51 no.2
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    • pp.45-49
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    • 2008
  • Cytosolic fructose-1,6-bisphosphatase (cFBPase) in plants is a key regulatory enzyme in the photosynthetic sucrose biosynthesis. Plant cFBPases, like the mammalian FBPases, are inhibited by adenosine 5'-monophosphate (AMP) and fructose-2,6-bisphosphate (Fru-2,6-$P_2$). In the mammalian FBPases, Lys112 and Tyr113 play important roles in the AMP binding. To understand roles of the corresponding residues, Lys115 and Tyr116, in pea cFBPase, the mutant cFBPases were generated by site-directed mutagenesis. The alterations of Lys115 to Gin and Tyr116 to Phe displayed small changes in $K_m$ and $K_i$ for Fru-2,6-$P_2$, indicating that the mutation causes minor effects on the enzyme catalysis and Fru-2,6-$P_2$ binding, whereas resulted in higher than 500-fold increase of $[AMP]_{0.5}$ compared with that of the wild-type enzyme. Results indicate the residues Lys115 and Tyr116 play important roles in the binding of AMP to the allosteric site of the pea cFBPase.

Chromosomal Mapping of the Gene Encoding Deoxycytidine-Cytidine Deaminase in Bacillus subtilis

  • Song, Bang-Ho;Jan Neuhard
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.512.2-512
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    • 1986
  • A mutant of bacillus subtilis with a defective cdd gene encoding deoxycytidine-cytidine deaminase(EC 3.5.4.5.) has been characterized genetically. The genetic lesion causing the altered deoxycytidine-cytidine deaminase, cdd, was mapped at 225 min on the linkage map of B.subtilis by AR9 transduction Transductional analysis of the cdd region established the gene order as trp-lys-dnaE-cdd-aroD. The cdd gene was linked 72% with the aroD and 20% with the lys.

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Sequence Analysis, Molecular Cloning and Restriction Mapping of Mitochondreal Genome of Domesticated Silkworm, Bombyx mori (누에 미토콘드리아 유전체의 제한효소 지도작성, 클로닝 및 염기서열 분석)

  • 이진성;성승현;김용성;서동상
    • Journal of Sericultural and Entomological Science
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    • v.42 no.1
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    • pp.14-23
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    • 2000
  • The mitochondrial genome of domesticated silkworm (Bombyx mori) was mapped with five restriction endonucleases (BamHI, EcoRI, HindIII, PstI and XbaI), the entire genome was cloned with HindIII and EcoRI. From the end sequencing results of 5$^1$and 3$^1$region for full genome set of eleven mitochondrial clones, the seven mitochondrial genes (NADH dehydrogenase 6, ATPase 6, ATPase 8, tRN $A^{Lys}$, tRN $A^{Asp}$, tRN $A^{Thr}$ and tRN $A^{Phe}$ of mori were identified on the basis of their nucleotide sequence homology. The nucleotide composition of NADH dehydrogenase 6 was heavily biased towards adenine and thymine, which accounted for 87.76%. On basis of the sequence similarity with published tRNA genes from six insect species, the tRN $A^{Lys}$, tRN $A^{Asp}$ and tRN $A^{Thr}$ were showed stable canonical clover-leaf tRNA structures with acceptible anticodons. However, both the DHU and T$\psi$C arms of tRN $A^{Phe}$ could not form any stable stem-loop structure. The two overlapping gene pairs (tRN $A^{Lys}$ -tRN $A^{ASP}$ and ATPase8-ATPase6) were found from our sequencing results. The genes are encoded on the same strad. ATPase8 and ATPase6 overlaps (ATGATAA) which are a single example of overlapping events between abutted protein-coding genes are common, and there is evidence that the two proteins are transcribed from a single bicistronic message by initiation at 5$^1$terminal start site for ATPase8 and at an internal start site for ATPase6. Ultimately, this result will provide assistance in designing oligo-nucleotides for PCR amplification, and sequencing the specific mitochondrial genes for phylogenetics of geographic races, genetically improved silkworm strains and wild silkworm (mandarina) which is estimated as ancestal of domesticated silkworm.sticated silkworm.

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Effect of In ovo Injection of Critical Amino Acids on Pre- and Post-hatch Growth, Immunocompetence and Development of Digestive Organs in Broiler Chickens

  • Bhanja, S.K.;Mandal, A.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.524-531
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    • 2005
  • Two experiments were conducted to standardize in ovo injection of amino acids (AA) and to evaluate the effect of in ovo injection of limiting AA(s) on pre and post hatch growth performance, immune response and development of digestive organs. Combinations of essential and non-essential amino acids (Lys+Arg, Lys+Met+Cys, Thr+Gly+Ser, Ile+Leu+Val and Gly+Pro) were injected into 50 eggs in each treatment group at 14 d of embryonic age. Standardization of injection site, needle length and embryonic age revealed that when AA were injected in to the broad end of the egg with a 11 mm needle and at the narrow end with a 24 mm needle both at the 7$^{th}$ and 14$^{th}$ d of incubation there was poor hatchability. However, better hatchability was recorded when the AA were injected in the narrow end of the egg with a 11 mm needle and in the broad end with a 24 mm needle on the 14$^{th}$ d of incubation. The chick to egg weight ratio was higher (p<0.018) when AA were injected on the 14$^{th}$ d of incubation. When a combination of amino acids were injected a 63.6 or 63.2 g difference in body weight of bird at 21 d was recorded between uninjected control and Ile+Leu+Val or Gly+Pro group, respectively. Higher feed intake (p<0.047) was recorded in the AA injected groups and feed conversion ratio (FCR) was numerically better in Gly+Pro, Lys+Met+Cys AA injected groups than in the uninjected control. Significantly higher immune response to cell mediated (p<0.033) and humoral (p<0.002) immunity was observed in in ovo amino acid injected birds, especially in Lys+Met+Cys, Thr+Gly+Ser or Ile+leu+Val groups. The digestive organ weights at 21 d did not differ between specific AA injected groups and the uninjected control. In ovo injected amino acids may act as immunomodulators and their role in gastrointestinal development needs further research.

DNA Repair Gene ERCC1 and XPD Polymorphisms Predict Glioma Susceptibility and Prognosis

  • Chen, Da-Qing;Yao, Dong-Xiao;Zhao, Hong-Yang;Yang, Shu-Juan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2791-2794
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    • 2012
  • Aims: We conducted a case-control study in a Chinese population to clarify the association between polymorphisms in ERCC1 and XPD and susceptibility and survival of glioma. Methods: A total of 393 cases and 410 controls were selected from March 2007 to December 2011. Genotyping of ERCC1 and XPD was conducted by TaqMan assays using the ABI Prism 7911HT Sequence Detection System. All analyses were performed using the STATA statistical package. Results: Polymorphisms in ERCC1 118C/T, ERCC1 8092C/A and XPD Asp312Asn showed no statistically significant difference between glioma cases and controls. However, individuals with the XPD 751Gln/Gln genotype had an increased risk of developing glioma compared with those with the Lys/Lys genotype (adjusted OR=1.64, 95% CI: 1.06-2.89). The ERCC1 118T/T genotype was associated with significantly higher median survival than the ERCC1 C/C genotype (HR=0.67, 95%CI=0.35-0.96). In addition, individuals with XPD 751Gln/Gln had a lower median survival time than XPD Lys/Lys carriers (HR=0.54, 95%CI=0.37-0.93). Conclusion: In conclusion, we observed that the XPD 751Gln/Gln genotype is associated with glioma susceptibility, and ERCC1 118 T/T and XPD 751Gln/Gln genotypes confer a significantly better prognosis.