• Title/Summary/Keyword: Chitin synthase

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Characterization and Phylogenetic Analysis of Chitin Synthase Genes from the Genera Sporobolomyces and Bensingtonia subrorea

  • Nam, Jin-Sik
    • Korean Journal of Environmental Biology
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    • v.23 no.4
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    • pp.335-342
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    • 2005
  • We cloned seven genes encoding chitin synthases (CHSs) by PCR amplification from genomic DNAs of four strains of the genus Sporobolomyces and of Bensingtonia subrosea using degenerated primers based on conserved regions of the CHS genes. Though amino acid sequences of these genes were shown similar as 176 to 189 amino acids except SgCHS2, DNA sequences were different in size, which was due to various introns present in seven fragments. Alignment and phylogenetic analysis of their deduced amino acid sequences together with the reported CHS genes of basidiomycetes separated the sequences into classes I, II and III. This analysis also permitted the classification of isolated CHSs; SgCHS1 belongs to class I, BsCHS1, SaCHS1, SgCHS2, SpgCHS1, and SsCHS1 belong to class II, and BsCHS2 belongs to class III. The deduced amino acid sequences involving in class II that were discovered from five strains were also compared with those of other basidiomycetes by CLUSTAL X program. The bootstrap analysis and phylogenetic tree by neighbor-joining method revealed the taxonomic and evolutionary position for four strains of the genus Sporobolomyces and for Bensingtonia subrosea which agreed with the previous classification. The results clearly showed that CHS fragments could be used as a valuable key for the molecular taxonomic and phylogenetic studies of basidiomycetes.

Role of LAMMER Kinase in Cell Wall Biogenesis during Vegetative Growth of Aspergillus nidulans

  • Choi, Yu Kyung;Kang, Eun-Hye;Park, Hee-Moon
    • Mycobiology
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    • v.42 no.4
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    • pp.422-426
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    • 2014
  • Depending on the acquisition of developmental competence, the expression of genes for ${\beta}$-1,3-glucan synthase and chitin synthase was affected in different ways by Aspergillus nidulans LAMMER kinase. LAMMER kinase deletion, ${\Delta}lkhA$, led to decrease in ${\beta}$-1,3-glucan, but increase in chitin content. The ${\Delta}lkhA$ strain was also resistant to nikkomycin Z.

Phenylpropanoids from Myristica fragrans Houtt (육두구(Myristica fragrans Houtt)로부터 Phenylpropanoid의 분리)

  • Song, Myoung-Chong;Ahn, Eun-Mi;Bang, Myun-Ho;Kim, Se-Young;Rho, Yeong-Deok;Kwon, Byuong-Mog;Lee, Hyun-Sun;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.47 no.3
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    • pp.366-369
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    • 2004
  • Myristica fragrans Houtt were extracted in 80% aq. MeOH and solvent fractionated sing $CHCl_3$, EtOAc, n-BuOH and water, successively. The n-BuOH fraction gave three phenylpropanoids through application of silica gel column chromatographies. The chemical structures of the phenylpropanoids were determined by the interpretation of several spectral data, including NMR and MS as meso-dihydroguaiaretic acid (1), nectandrin B (2) and syringin methyl ether (3). Compound 1, which was first isolated from this plant by authors, showed inhibitory activities with $60.0{\pm}2.1%\;(100\;{\mu}g/ml),\;42.6{\pm}0.9%\;(140\;{\mu}g/ml)\;and\;12.2{\pm}0.2%\;(200\;{\mu}g/ml)$ on ACAT(acyl-CoA:Cholesterol Acyltransferase), chitin synthase III and HMG-CoA reductase (3-hydroxy-3-methylglutaryl coenzyme A reductase), respectively. Compound 3 showed inhibitory activities with $27.2{\pm}0.9%\;(100\;{\mu}g/ml),\;45.5{\pm}0.8%\;(200\;{\mu}g/ml)$ on ACAT and chitin synthase III.

Effect of KGD1 Deletion on Cell Wall Biogenesis in Saccharomyces cerevisiae (Saccharomyces cerevisiae의 KGD1 유전자 결손이 세포벽 생합성에 미치는 영향)

  • Kim, Sung-Woo;Ahn, Ki-Woong;Park, Yun-Hee;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.29-33
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    • 2010
  • KGD1 gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. We performed the gene disruption experiment to characterize the function of KGD1 gene, which encodes $\beta$-ketoglutarate dehydrogenase, in cell wall biosynthesis. The disruption of KGD1 showed the decreased growth rate, the increase of chitin synthases activity, alterations in cell wall composition, and increase of susceptibility to cell wall inhibitors such as Calcofluor white and Nikkomycin Z. These results suggested that KGD1 might be involved in cell wall biogenesis, especially the biosynthesis of $\beta$-1,6-glucan and chitin in S. cerevisaie.

Identification of a Domain in Yeast Chitin Synthase 3 Required for Biogenesis of Chitin Ring, But Not Cellular Chitin Synthesis

  • Park Hyun-Sook;Park Mee-Hyun;Kim Chi-Hwa;Woo Jeeun;Lee Jee-Yeon;Kim Sung-Uk;Choi Wonja
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.39-45
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    • 2000
  • It hab been proposed that CHS3-mediated chitin synthesis during the vegitative cell cycle is regulated by CHS4. To investigate direct protein-protein interaction between their coding products, we used yeast two hybrid system and found that a domain of Chs3p was responsible for interaction with Chs4p. This domain, termed MIRC3-4 (maximum interacting region of chs3p with chs4p), spans from 647 to 700 residues. It is well conserved among CHS3 homologs of various fungi such as Candida albicans, Emericella nidulans, Neurospora crassa, Magnaporthe grisea, Ustilago maydis, Glomus versiforme, Exophiala dermatitidis, Rhizopus microsporus. A series of mutaion in the MIRC3-4 resulted in no appearance of chitin ring at the early G 1 phase but did not affect chitin synthesis in the cell wall after cytokinesis. Absence of chitin ring could be caused either by delocalization of Chs3p to the septum or by improper interaction with Chs4p. To discriminate those two, not mutually exclusive, alternatives, mutants cells were immunostained with Chs3p-specific antibody. Some exhibited localization of chs3p to the septum, while others failed. These results indicate that simultaneous localization and activation Chs3p by Chs4p is required for chitin ring synthesis.

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The Determination of Chitin Synthases by Varying pH and Divalent Cations in Candida albicans

  • Choi, Won-Ja
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.613-617
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    • 1998
  • The Chsl, Chs2, and Chs3 activities of a pathogenic fungus, Candida albicans, perform the same biochemical reactions, but exert different functions. Therefore, the determination of each enzyme activity is important. The three chitin synthases differ in their optimal pH and the effect of divalent cations as either stimulatory or inhibitory factors. The CAChsl, CAChs2, and CAChs3 activities are optimal at pH 7.5, 6.5, and 8.5, respectively. $Co^{2+} stimulates CAChsl and CAChs3, but inhibits CAChs2. $Ni^{2+}$ inhibits CAChsl and CAChs2 with little effect on CAChs3. $Mg^{2+}$ stimulates CAChs2 and CAChs3, but hardly affects CAChsl. These characteristics are similar to those of the Saccharomyces cerevisiae enzymes except in degree. The sensitivity against $Ni^{2+}$ of CAChsl is higher than that of CAChs2, whereas the reverse is true in S. cerevisiae. Metal dependence of chitin synthases in C. albicans is less marked than that in S. cerevisiae, except for CAChs2. The activities of CAChsl and CAChs3 from EDTA-treated membranes were increased 1.5 fold, while that of CAChs2 was stimulated 7 fold in the presence of divalent cations. These results could provide new criteria for screening systems of antifungal agents.

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