scholarly journals Purification of arginase from Aspergillus nidulans.

1994 ◽  
Vol 41 (4) ◽  
pp. 467-471 ◽  
Author(s):  
A Dzikowska ◽  
J P Le Caer ◽  
P Jonczyk ◽  
P Wëgleński
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Neurospora Crassa ◽  
Molecular Mass ◽  
Nitrogen Source ◽  
Aspergillus Nidulans ◽  
Sole Nitrogen Source ◽  
Mass Ratio ◽  
Conserved Domains ◽  
Native Molecular Mass ◽  
High Degree

Arginase (EC 3.5.3.1) of Aspergillus nidulans, the enzyme which enables the fungus to use arginine as the sole nitrogen source was purified to homogeneity. Molecular mass of the purified arginase subunit is 40 kDa and is similar to that reported for the Neurospora crassa (38.3 kDa) and Saccharomyces cerevisiae (39 kDa) enzymes. The native molecular mass of arginase is 125 kDa. The subunit/native molecular mass ratio suggests a trimeric form of the protein. The arginase protein was cleaved and partially sequenced. Two out of the six polypeptides sequenced show a high degree of homology to conserved domains in arginases from other species.

scholarly journals Improved Anaerobic Use of Arginine by Saccharomyces cerevisiae

2003 ◽  
Vol 69 (3) ◽  
pp. 1623-1628 ◽  
Author(s):  
Olga Martin ◽  
Marjorie C. Brandriss ◽  
Gisbert Schneider ◽  
Alan T. Bakalinsky
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nitrogen Source ◽  
Genetically Modified ◽  
Cell Mass ◽  
Sole Nitrogen Source ◽  
Mitochondrial Targeting ◽  
Arginine Catabolism

ABSTRACT Anaerobic arginine catabolism in Saccharomyces cerevisiae was genetically modified to allow assimilation of all four rather than just three of the nitrogen atoms in arginine. This was accomplished by bypassing normal formation of proline, an unusable nitrogen source in the absence of oxygen, and causing formation of glutamate instead. A pro3 ure2 strain expressing a PGK1 promoter-driven PUT2 allele encoding Δ1-pyrroline-5-carboxylate dehydrogenase lacking a mitochondrial targeting sequence produced significant cytoplasmic activity, accumulated twice as much intracellular glutamate, and produced twice as much cell mass as the parent when grown anaerobically on limiting arginine as sole nitrogen source.

scholarly journals Homologous npdGI Genes in 2,4-Dinitrophenol- and 4-Nitrophenol-Degrading Rhodococcus spp

2003 ◽  
Vol 69 (5) ◽  
pp. 2748-2754 ◽  
Author(s):  
Gesche Heiss ◽  
Natalie Trachtmann ◽  
Yoshikatsu Abe ◽  
Masahiro Takeo ◽  
Hans-Joachim Knackmuss
Keyword(s):  
Heterologous Expression ◽  
Nitrogen Source ◽  
Phylogenetic Analyses ◽  
Sole Nitrogen Source ◽  
Sequence Analyses ◽  
Conserved Domains ◽  
Meisenheimer Complexes ◽  
Additional Group ◽  
The Family ◽  
Sequence Similarities

ABSTRACT Rhodococcus (opacus) erythropolis HL PM-1 grows on 2,4,6-trinitrophenol or 2,4-dinitrophenol (2,4-DNP) as a sole nitrogen source. The NADPH-dependent F420 reductase (NDFR; encoded by npdG) and the hydride transferase II (HTII; encoded by npdI) of the strain were previously shown to convert both nitrophenols to their respective hydride Meisenheimer complexes. In the present study, npdG and npdI were amplified from six 2,4-DNP degrading Rhodococcus spp. The genes showed sequence similarities of 86 to 99% to the respective npd genes of strain HL PM-1. Heterologous expression of the npdG and npdI genes showed that they were involved in 2,4-DNP degradation. Sequence analyses of both the NDFRs and the HTIIs revealed conserved domains which may be involved in binding of NADPH or F420. Phylogenetic analyses of the NDFRs showed that they represent a new group in the family of F420-dependent NADPH reductases. Phylogenetic analyses of the HTIIs revealed that they form an additional group in the family of F420-dependent glucose-6-phosphate dehydrogenases and F420-dependent N 5,N 10-methylenetetrahydromethanopterin reductases. Thus, the NDFRs and the HTIIs may each represent a novel group of F420-dependent enzymes involved in catabolism.

scholarly journals The effect of thiourea on ureide metabolism in Neurospora crassa

1973 ◽  
Vol 136 (3) ◽  
pp. 749-755 ◽  
Author(s):  
Jasti Nirmala ◽  
Killampalli Sivarama Sastry
Keyword(s):  
Neurospora Crassa ◽  
Nitrogen Source ◽  
Type Strain ◽  
Wild Type Strain ◽  
Inorganic Nitrogen ◽  
Wild Strain ◽  
Sole Nitrogen Source ◽  
Wild Type ◽  
Selective Toxicity ◽  
In The Wild

The wild-type strain of Neurospora crassa Em 5297a can utilize allantoin as a sole nitrogen source. The pathway of allantoin utilization is via its conversion into allantoic acid and urea, followed by the breakdown of urea to ammonia. This is shown by the inability of the urease-less mutant, N. crassa 1229, to grow on allantoin as a sole nitrogen source and by the formation of allantoate and urea by pre-formed mycelia of this mutant. In the wild strain (Em 5297a) thiourea is tenfold more toxic on an allantoin medium than on an inorganic nitrogen medium; allantoin as well as urea counteract thiourea toxicity in the allantoin nitrogen medium. This selective toxicity of thiourea for the mould utilizing allantoin nitrogen does not, however, result in an impairment of allantoin uptake, allantoinase activity or the formation of urea from allantoin. The only process affected by thiourea is the synthesis of urease; urea antagonizes this effect of thiourea in N. crassa.

scholarly journals CYTOCHEMICAL DETECTION OF HYDROLASES IN FUNGUS CELLS III. ARYL SULFATASE

1974 ◽  
Vol 22 (3) ◽  
pp. 183-188 ◽  
Author(s):  
JÜRGEN REISS
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Enzyme Activity ◽  
Neurospora Crassa ◽  
Aspergillus Nidulans ◽  
Aspergillus Oryzae ◽  
Coupling Agent ◽  
Mycelial Fungi ◽  
Aryl Sulfatase ◽  
Negative Results ◽  
Coupling Procedure

In the cells of Aspergillus oryzae, Aspergillus nidulans, Neurospora crassa and Saccharomyces cerevisiae, aryl sulfatase can be demonstrated by incubation in a medium containing 6-bromo-2-naphthylsulfate as substrate and fast garnet GBC as coupling agent. Controls confirm the specificity of the reaction. Other incubation solutions (two Gomori media and a simultaneous coupling procedure with 8-hydroxyquinoline sulfate as substrate) gave negative results or reaction pictures equally to those in substrate-free control. The possible reasons for this are discussed. In the mycelial fungi the strongest enzyme activity is located in the most intensely metabolizing parts: tips of the hyphae and the differentiating parts of the conidiophores. The reaction granules in all four fungi are possibly identical with lysosomes.

Regulation of mannitol-1-phosphate dehydrogenase in Aspergillus nidulans

1975 ◽  
Vol 21 (1) ◽  
pp. 99-101 ◽  
Author(s):  
Oliver Hankinson ◽  
David J. Cove
Keyword(s):  
Nitrogen Source ◽  
Aspergillus Nidulans ◽  
Sodium Nitrate ◽  
Sole Nitrogen Source

When Aspergillus nidulans is grown with urea as sole nitrogen source it possesses about sixtold higher activity of mannitol-1-phosphate dehydrogenase than when it is grown with both urea and sodium nitrate.

scholarly journals Inositol trisphosphate metabolism in Saccharomyces cerevisiae: identification, purification and properties of inositol 1,4,5-trisphosphate 6-kinase

1994 ◽  
Vol 302 (3) ◽  
pp. 709-716 ◽  
Author(s):  
F Estevez ◽  
D Pulford ◽  
M J Stark ◽  
A N Carter ◽  
C P Downes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Mass ◽  
Human Erythrocyte ◽  
Kinase Activity ◽  
Erythrocyte Ghosts ◽  
Inositol Polyphosphates ◽  
Step Process ◽  
Native Molecular Mass ◽  
Purification And Properties ◽  
Inositol 1,4,5 Trisphosphate

Ins(1,4,5)P3 metabolism was examined in Saccharomyces cerevisiae extracts. S. cerevisiae contains readily detectable Ins(1,4,5)P3 kinase activity that is predominantly soluble, but phosphomonoesterase activity acting on Ins(1,4,5)P3 was not detected in either soluble or particulate preparations from this organism. We have purified the kinase activity approximately 685-fold in a rapid four-step process, and obtained a stable preparation. The enzyme has an apparent native molecular mass of approximately 40 kDa, and displays Michaelis-Menten kinetics with respect to its two substrates, ATP and Ins(1,4,5)P3. The Km for ATP was 2.1 mM, and that for Ins(1,4,5)P3 was 7.1 microM. The enzyme appeared to be the first step in the conversion of Ins(1,4,5)P3 into an InsP5, and the partially purified preparation contained another activity that converted the InsP4 product into an InsP5. The InsP4 product of the partially purified kinase was not metabolized by human erythrocyte ghosts and co-chromatographed with an Ins(3,4,5,6)P4 [L-Ins(1,4,5,6)P4] standard, identifying it as D-Ins(1,4,5,6)P4. The yeast enzyme is thus an Ins(1,4,5)P3 6-kinase. This activity may be an important step in the production of inositol polyphosphates such as InsP5 and InsP6 in S. cerevisiae.

scholarly journals Genetics of the synthesis of serine from glycine and the utilization of glycine as sole nitrogen source by Saccharomyces cerevisiae.

Genetics ◽  
1995 ◽  
Vol 140 (4) ◽  
pp. 1213-1222 ◽  
Author(s):  
D A Sinclair ◽  
I W Dawes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nitrogen Source ◽  
Single Gene ◽  
Glycine Decarboxylase ◽  
Multienzyme Complex ◽  
Sole Nitrogen Source ◽  
Yeast Strains ◽  
Complementation Groups ◽  
Lipoamide Dehydrogenase ◽  
Serine Biosynthesis

Abstract Saccharomyces cerevisiae can grow on glycine as sole nitrogen source and can convert glycine to serine via the reaction catalyzed by the glycine decarboxylase multienzyme complex (GDC). Yeast strains with mutations in the single gene for lipoamide dehydrogenase (lpd1) lack GDC activity, as well as the other three 2-oxoacid dehydrogenases dependent on this enzyme. The LPD1 gene product is also required for cells to utilize glycine as sole nitrogen source. The effect of mutations in LPD1 (L-subunit of GDC), SER1 (synthesis of serine from 3-phosphoglycerate), ADE3 (cytoplasmic synthesis of one-carbon units for the serine synthesis from glycine), and all combinations of each has been determined. The results were used to devise methods for isolating mutants affected either in the generation of one-carbon units from glycine (via GDC) or subsequent steps in serine biosynthesis. The mutants fell into six complementation groups (gsd1-6 for defects in conversion of glycine to serine). Representatives from three complementation groups were also unable to grow on glycine as sole nitrogen source (gsd1-3). Assays of the rate of glycine uptake and decarboxylation have provided insights into the nature of the mutations.

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