The dimorphic yeast Yarrowia lipolytica possesses an atypical phosphofructokinase: characterization of the enzyme and its encoding gene

Microbiology ◽  
2005 ◽  
Vol 151 (5) ◽  
pp. 1465-1474 ◽  
Author(s):  
Carmen-Lisset Flores ◽  
Oscar H. Martínez-Costa ◽  
Valentina Sánchez ◽  
Carlos Gancedo ◽  
Juan J. Aragón
Keyword(s):  
Yarrowia Lipolytica ◽  
Carbon Sources ◽  
Hill Coefficient ◽  
Single Type ◽  
Kinetic Characteristics ◽  
Gene Encoding ◽  
Dimorphic Yeast ◽  
Encoding Gene ◽  
Cooperative Kinetics

The phosphofructokinase from the non-conventional yeast Yarrowia lipolytica (YlPfk) was purified to homogeneity, and its encoding gene isolated. YlPfk is an octamer of 869 kDa composed of a single type of subunit, and shows atypical kinetic characteristics. It did not exhibit cooperative kinetics for fructose 6-phosphate (Hill coefficient, h 1·1; S 0·5 52 μM), it was inhibited moderately by MgATP (K i 3·5 mM), and it was strongly inhibited by phosphoenolpyruvate (K i 61 μM). Fructose 2,6-bisphosphate did not activate the enzyme, and AMP and ADP were also without effect. The gene YlPFK1 has no introns, and encodes a putative protein of 953 aa, with a molecular mass consistent with the subunit size found after purification. Disruption of the gene abolished growth in glucose and Pfk activity, while reintroduction of the gene restored both properties. This indicates that Y. lipolytica has only one gene encoding Pfk, and supports the finding that the enzyme consists of identical subunits. Glucose did not interfere with growth of the Ylpfk1 disruptant in permissive carbon sources. The unusual kinetic characteristics of YlPfk, and the intracellular concentrations of glycolytic intermediates during growth in glucose, suggest that YlPfk may play an important role in the regulation of glucose metabolism in Y. lipolytica, different from the role played by the enzyme in Saccharomyces cerevisiae.

Functional characterization of extracellular chitinase encoded by the YlCTS1 gene in a dimorphic yeast Yarrowia lipolytica

2014 ◽  
Vol 52 (4) ◽  
pp. 284-291 ◽  
Author(s):  
Jeong-Nam Park ◽  
Chang Pyo Han ◽  
Dong-Jik Lee ◽  
Seon Ah Cheon ◽  
Hyun Ah Kang
Keyword(s):  
Yarrowia Lipolytica ◽  
Functional Characterization ◽  
Dimorphic Yeast ◽  
Yeast Yarrowia Lipolytica

scholarly journals Genetic and Biochemical Characterization of FRI-1, a Carbapenem-Hydrolyzing Class A β-Lactamase from Enterobacter cloacae

2015 ◽  
Vol 59 (12) ◽  
pp. 7420-7425 ◽  
Author(s):  
Laurent Dortet ◽  
Laurent Poirel ◽  
Samia Abbas ◽  
Saoussen Oueslati ◽  
Patrice Nordmann
Keyword(s):  
Biochemical Characterization ◽  
Enterobacter Cloacae ◽  
Conjugative Plasmid ◽  
Lower Sensitivity ◽  
Gene Encoding ◽  
Content Type ◽  
Class A ◽  
Encoding Gene ◽  
Lactamase Gene

ABSTRACTAnEnterobacter cloacaeisolate was recovered from a rectal swab from a patient hospitalized in France with previous travel to Switzerland. It was resistant to penicillins, narrow- and broad-spectrum cephalosporins, aztreonam, and carbapenems but remained susceptible to expanded-spectrum cephalosporins. Whereas PCR-based identification of the most common carbapenemase genes failed, the biochemical Carba NP test II identified an Ambler class A carbapenemase. Cloning experiments followed by sequencing identified a gene encoding a totally novel class A carbapenemase, FRI-1, sharing 51 to 55% amino acid sequence identity with the closest carbapenemase sequences. However, it shared conserved residues as a source of carbapenemase activity. Purified β-lactamase FRI-1 hydrolyzed penicillins, aztreonam, and carbapenems but spared expanded-spectrum cephalosporins. The 50% inhibitory concentrations (IC50s) of clavulanic acid and tazobactam were 10-fold higher than those found forKlebsiella pneumoniaecarbapenemase (KPC), IMI, and SME, leading to lower sensitivity of FRI-1 activity to β-lactamase inhibitors. TheblaFRI-1gene was located on a ca. 110-kb untypeable, transferable, and non-self-conjugative plasmid. A putative LysR family regulator-encoding gene at the 5′ end of the β-lactamase gene was identified, leading to inducible expression of theblaFRI-1gene.

scholarly journals Cloning and Expression of the algL Gene, Encoding the Azotobacter chroococcum Alginate Lyase: Purification and Characterization of the Enzyme

1999 ◽  
Vol 181 (5) ◽  
pp. 1409-1414 ◽  
Author(s):  
Ana Peciña ◽  
Alberto Pascual ◽  
Antonio Paneque
Keyword(s):  
Gene Sequence ◽  
Azotobacter Vinelandii ◽  
Alginate Lyase ◽  
Azotobacter Chroococcum ◽  
Open Reading Frame ◽  
Cloning And Expression ◽  
Gene Encoding ◽  
Reading Frame ◽  
Encoding Gene

ABSTRACT The alginate lyase-encoding gene (algL) ofAzotobacter chroococcum was localized to a 3.1-kbEcoRI DNA fragment that revealed an open reading frame of 1,116 bp. This open reading frame encodes a protein of 42.98 kDa, in agreement with the value previously reported by us for this protein. The deduced protein has a potential N-terminal signal peptide that is consistent with its proposed periplasmic location. The analysis of the deduced amino acid sequence indicated that the gene sequence has a high homology (90% identity) to the Azotobacter vinelandii gene sequence, which has very recently been deposited in the GenBank database, and that it has 64% identity to the Pseudomonas aeruginosa gene sequence but that it has rather low homology (15 to 22% identity) to the gene sequences encoding alginate lyase in other bacteria. The A. chroococcum AlgL protein was overproduced in Escherichia coli and purified to electrophoretic homogeneity in a two-step chromatography procedure on hydroxyapatite and phenyl-Sepharose. The kinetic and molecular parameters of the recombinant alginate lyase are similar to those found for the native enzyme.

scholarly journals A Rac Homolog Is Required for Induction of Hyphal Growth in the Dimorphic Yeast Yarrowia lipolytica

2000 ◽  
Vol 182 (9) ◽  
pp. 2376-2386 ◽  
Author(s):  
Cleofe A. R. Hurtado ◽  
Jean-Marie Beckerich ◽  
Claude Gaillardin ◽  
Richard A. Rachubinski
Keyword(s):  
Yarrowia Lipolytica ◽  
Blot Analysis ◽  
Zinc Finger Protein ◽  
Essential Gene ◽  
Hyphal Growth ◽  
Fungal Species ◽  
Gene Encoding ◽  
Adverse Conditions ◽  
Acid Protein ◽  
Dimorphic Yeast

ABSTRACT Dimorphism in fungi is believed to constitute a mechanism of response to adverse conditions and represents an important attribute for the development of virulence by a number of pathogenic fungal species. We have isolated YlRAC1, a gene encoding a 192-amino-acid protein that is essential for hyphal growth in the dimorphic yeast Yarrowia lipolytica and which represents the first Rac homolog described for fungi. YlRAC1 is not an essential gene, and its deletion does not affect the ability to mate or impair actin polarization in Y. lipolytica. However, strains lacking functional YlRAC1 show alterations in cell morphology, suggesting that the function of YlRAC1 may be related to some aspect of the polarization of cell growth. Northern blot analysis showed that transcription of YlRAC1 increases steadily during the yeast-to-hypha transition, while Southern blot analysis of genomic DNA suggested the presence of severalRAC family members in Y. lipolytica. Interestingly, strains lacking functional YlRAC1 are still able to grow as the pseudohyphal form and to invade agar, thus pointing to a function for YlRAC1 downstream of MHY1, a previously isolated gene encoding a C2H2-type zinc finger protein with the ability to bind putative stress response elements and whose activity is essential for both hyphal and pseudohyphal growth in Y. lipolytica.

Molecular characterization of acidic peptide:N-glycanase from the dimorphic yeast Yarrowia lipolytica

2014 ◽  
Vol 157 (1) ◽  
pp. 35-43 ◽  
Author(s):  
Kyung Jin Lee ◽  
Jin Young Gil ◽  
Sang-Yoon Kim ◽  
Ohsuk Kwon ◽  
Kisung Ko ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Yarrowia Lipolytica ◽  
Dimorphic Yeast ◽  
Yeast Yarrowia Lipolytica

The isolation and characterization of the pyruvate kinase-encoding gene from the yeast Yarrowia lipolytica

Gene ◽  
1994 ◽  
Vol 140 (1) ◽  
pp. 141-143 ◽  
Author(s):  
C.A. Strick ◽  
L.C. James ◽  
M.M. O'Donnell ◽  
M.G. Gollaher ◽  
A.E. Franke
Keyword(s):  
Pyruvate Kinase ◽  
Yarrowia Lipolytica ◽  
Isolation And Characterization ◽  
Encoding Gene ◽  
Yeast Yarrowia Lipolytica

scholarly journals Molecular Characterization of a TEM-21 β-Lactamase in a Clinical Isolate of Morganella morganii

1998 ◽  
Vol 42 (8) ◽  
pp. 2125-2127 ◽  
Author(s):  
F. Tessier ◽  
C. Arpin ◽  
A. Allery ◽  
C. Quentin
Keyword(s):  
Nucleotide Sequence ◽  
Molecular Characterization ◽  
Clinical Isolate ◽  
Morganella Morganii ◽  
Gene Encoding ◽  
Extended Spectrum ◽  
Encoding Gene

ABSTRACT A clinical isolate of Morganella morganii, with reduced susceptibility to expanded-spectrum cephalosporins and aztreonam, was found to produce an extended-spectrum β-lactamase with a pI of 6.4. The nucleotide sequence of the encoding gene was that of the gene encoding TEM-21. This is the first molecular characterization of an extended-spectrum β-lactamase in M. morganii.

Expression regulation of the endochitinase-encoding genesechi44from the mycoparasiteStachybotrys elegans

2006 ◽  
Vol 52 (11) ◽  
pp. 1103-1109 ◽  
Author(s):  
D C Morissette ◽  
P Seguin ◽  
S H Jabaji-Hare
Keyword(s):  
Real Time ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Rt Pcr ◽  
Gene Encoding ◽  
Carbon And Nitrogen ◽  
Cyclical Pattern ◽  
Polymerase Chain ◽  
Encoding Gene ◽  
High Concentrations

The regulation of the gene encoding the extracellular chitinase sechi44 produced by the mycoparasite Stachybotrys elegans was studied using real-time quantitative reverse-transcription polymerase chain reaction. Alteration of sechi44 expression was observed when S. elegans was in interaction with its host, Rhizoctonia solani, and also when the mycoparasite was grown on minimal media amended with different carbon and nitrogen sources. Direct contact with R. solani leading to mycoparasitism significantly up-regulated the expression of sechi44, although the analysis showed that sechi44 was constitutively expressed but at substantially lower levels. In addition, the study of sechi44 over 12 days showed that its expression followed a cyclical pattern with peaks every 2 days, which suggests that this gene has a role not only in mycoparasitism but also in growth. The addition of external carbon sources, such as N-acetylglucosamine, chitin, and R. solani cell wall (simulated mycoparasitism), triggered an increase in the expression of sechi44, which varied with time and carbon source. Among the carbon sources examined, N-acetylglucosamine induced the highest increase in sechi44 transcript levels. The addition of high concentrations of glucose and ammonium triggered a decrease of sechi44 expression, suggesting that sechi44 is subject to glucose and ammonium repression.Key words: mycoparasitism, Stachybotrys elegans, endochitinase-encoding gene, sechi44, real-time RT–PCR.

scholarly journals Characterization of Saccharomyces cerevisiae deficient in expression of phospholipase D

1996 ◽  
Vol 314 (1) ◽  
pp. 15-19 ◽  
Author(s):  
Krishna M. ELLA ◽  
Joseph W. DOLAN ◽  
Chen QI ◽  
Kathryn E. MEIER
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Phospholipase D ◽  
Gene Disruption ◽  
Carbon Sources ◽  
Gene Encoding ◽  
Sequence Identity ◽  
One Step

A gene encoding phospholipase D (PLD) in Saccharomyces cerevisiae was identified. The 195 kDa product of PLD1 has 24% overall sequence identity with a plant PLD. Expression of yeast PLD activity was eliminated by one-step gene disruption. Yeast haploids lacking PLD activity were deficient in growth on non-fermentable carbon sources. Diploids lacking expression of PLD1 were unable to sporulate.

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