The biosynthesis of glucosamine

Leloir, Luis F.; Cardini, Carlos E.

doi:10.1016/0006-3002(53)90119-x

Cited by 186 publications

(35 citation statements)

References 16 publications

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“…Tubes were incubated at 30°C for 30 min, and heating at 100°C for 2 min terminated the reaction. Amounts of GlcN-6-P formed were determined by a modified MorganElson procedure (29). Portions (10 l) of the reaction mixture were transferred to PCR strip tubes, and 4 l of 5% acetic anhydride in acetone was added to each tube and incubated for 3 min at room temperature, followed by addition of 14 l of 0.33 M potassium tetraborate, pH 9.0, and incubation at 99.9°C for 10 min in a thermocycler (Techne Inc.).…”

Section: Methodsmentioning

confidence: 99%

Chitin Synthesis in Saccharomyces cerevisiae in Response to Supplementation of Growth Medium withGlucosamine and Cell WallStress

et al. 2003

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In Saccharomyces cerevisiae most chitin is synthesized by Chs3p, which deposits chitin in the lateral cell wall and in the bud-neck region during cell division. We have recently found that addition of glucosamine (GlcN) to the growth medium leads to a three-to fourfold increase in cell wall chitin levels. We compared this result to the increases in cellular chitin levels associated with cell wall stress and with treatment of yeast with mating pheromone. Since all three phenomena lead to increases in precursors of chitin, we hypothesized that chitin synthesis is at least in part directly regulated by the size of this pool. This hypothesis was strengthened by our finding that addition of GlcN to the growth medium causes a rapid increase in chitin synthesis without any pronounced change in the expression of more than 6,000 genes monitored with Affymetrix gene expression chips. In other studies we found that the specific activity of Chs3p is higher in the total membrane fractions from cells grown in GlcN and from mutants with weakened cell walls. Sucrose gradient analysis shows that Chs3p is present in an inactive form in what may be Golgi compartments but as an active enzyme in other intracellular membrane-bound vesicles, as well as in the plasma membrane. We conclude that Chs3p-dependent chitin synthesis in S. cerevisiae is regulated both by the levels of intermediates of the UDP-GlcNAc biosynthetic pathway and by an increase in the activity of the enzyme in the plasma membrane.

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Section: Methodsmentioning

confidence: 99%

Chitin Synthesis in Saccharomyces cerevisiae in Response to Supplementation of Growth Medium withGlucosamine and Cell WallStress

et al. 2003

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“…The substrate for each chitin synthase isozyme is UDP-N-acetylglucosamine (UDP-GlcNAc), which is synthesized from fructose-6-phosphate by the Leloir pathway ( Fig. 17; Table 51) (466,529). The first pathway-specific enzyme is a ketol-isomerase that synthesizes glucosamine-6-phosphate (GlcN-6-P) and glutamate from glutamine and fructose-6-phosphate.…”

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confidence: 99%

Lessons from the Genome Sequence ofNeurospora crassa: Tracing the Path from Genomic Blueprint to Multicellular Organism

Borkovich

Alex

Yarden

et al. 2004

Microbiol Mol Biol Rev

598

618

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We present an analysis of over 1,100 of the ∼10,000 predicted proteins encoded by the genome sequence of the filamentous fungus Neurospora crassa. Seven major areas of Neurospora genomics and biology are covered. First, the basic features of the genome, including the automated assembly, gene calls, and global gene analyses are summarized. The second section covers components of the centromere and kinetochore complexes, chromatin assembly and modification, and transcription and translation initiation factors. The third area discusses genome defense mechanisms, including repeat induced point mutation, quelling and meiotic silencing, and DNA repair and recombination. In the fourth section, topics relevant to metabolism and transport include extracellular digestion; membrane transporters; aspects of carbon, sulfur, nitrogen, and lipid metabolism; the mitochondrion and energy metabolism; the proteasome; and protein glycosylation, secretion, and endocytosis. Environmental sensing is the focus of the fifth section with a treatment of two-component systems; GTP-binding proteins; mitogen-activated protein, p21-activated, and germinal center kinases; calcium signaling; protein phosphatases; photobiology; circadian rhythms; and heat shock and stress responses. The sixth area of analysis is growth and development; it encompasses cell wall synthesis, proteins important for hyphal polarity, cytoskeletal components, the cyclin/cyclin-dependent kinase machinery, macroconidiation, meiosis, and the sexual cycle. The seventh section covers topics relevant to animal and plant pathogenesis and human disease. The results demonstrate that a large proportion of Neurospora genes do not have homologues in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. The group of unshared genes includes potential new targets for antifungals as well as loci implicated in human and plant physiology and disease

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“…The following chemical compounds were obtained from Sigma Chemical Co.: sodium salt of Gm-6-P (98% pure); trilithium salt of CoA (85% pure); protamine sulfate (Grade I); PHMB (sodium salt); and Gm hydrochloride. Ellman's reagent [5,5'-dithiobis-(2-nitrobenzoic acid)] for the determination of sulfhydryl groups was purchased from Aldrich Chemical Co. All other chemicals were obtained through other commercial sources.…”

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confidence: 99%

Partial Purification and Properties of d-Glucosamine 6-Phosphate N-Acetyltransferase from Phaseolus aureus

Vessal

Hassid²

1973

Plant Physiol.

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D-Glucosamine-6-P N-acetyltransferase (EC 2.3.1.4) from mung bean seeds (Phaseolus aureus) was purified 313-fold by protamine sulfate and isoelectric precipitation, ammonium sulfate and acetone fractionation, and CM Sephadex column chromatography. The partially purified enzyme was highly specific for D-glucosamine-6-P. Neither D-glucosamine nor Dgalactosamine could replace this substrate. The partially purified enzyme preparation was inhibited up to 50% by 2 X 102 M EDTA, indicating the requirement of a divalent cation. Among divalent metal ions tested, Mg' was required for maximum activity of the enzyme. Mn'+ and Zn'+ were inhibitory, while Co+ had no effect on the enzyme activity. The pH optimum of the enzyme in sodium acetate and sodium citrate buffers was found to be 5.2. The effect of Mg2' on the enzyme in sodium acetate and sodium citrate buffers was particularly noticeable in the range of optimum pH. Km values of 15.1 X 10-4 M and 7.1 X 10-4 M were obtained for D-glucosamine-6-P and acetyl CoA, respectively. The enzyme was completely inhibited by 1 X 10-4 M p-hydroxymercuribenzoate, and this inhibition was partially reversed by L-cysteine; indicating the presence of sulfhydryl groups at or near the active site of the enzyme.D-Glucosamine-6-P N-acetyltransferase (EC 2.3.1.4), which is on the pathway for the biosynthesis of uridine diphosphate N-acetyl-D-glucosamine, catalyzes the following reversible reaction:Gm-6-P' + acetyl-CoA --AcGm-6-P + CoA Leloir and Cardini (5) demonstrated the presence of an enzyme in the crude extracts of Neurospora crassa capable of acetylating either Gm or Gm-6-P in the presence of acetyl-CoA. Later, Brown (2) demonstrated that an extract of bakers' yeast was

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The biosynthesis of glucosamine

Cited by 186 publications

References 16 publications

Chitin Synthesis in Saccharomyces cerevisiae in Response to Supplementation of Growth Medium withGlucosamine and Cell WallStress

Chitin Synthesis in Saccharomyces cerevisiae in Response to Supplementation of Growth Medium withGlucosamine and Cell WallStress

Lessons from the Genome Sequence ofNeurospora crassa: Tracing the Path from Genomic Blueprint to Multicellular Organism

Partial Purification and Properties of d-Glucosamine 6-Phosphate N-Acetyltransferase from Phaseolus aureus

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