Expression and characterization of the 42 kDa chitinase of the biocontrol fungusMetarhizium anisopliaeinEscherichia coli

2003 ◽  
Vol 49 (11) ◽  
pp. 723-726 ◽  
Author(s):  
César Milton Baratto ◽  
Marcia Vanusa da Silva ◽  
Lucélia Santi ◽  
Luciane Passaglia ◽  
Irene Silveira Schrank ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Metarhizium Anisopliae ◽  
Entomopathogenic Fungus ◽  
Expression Vector ◽  
Hydrolytic Enzymes ◽  
Active Protein ◽  
Chitinase Genes

Albeit Metarhizium anisopliae is the best-characterized entomopathogenic fungus, the role of some hydrolytic enzymes during host cuticle penetration has not yet been established. Three chitinase genes (chit1, chi2, chi3) from Metarhizium have already been isolated. To characterize the chitinase coded by the chit1 gene, we expressed the active protein (CHIT42) in Escherichia coli using a T7-based promoter expression vector. The recombinant protein, CHIT42, is active against glycol chitin and synthetic N-acetylglucosamine (GlcNAc) dimer and tetramer substrates. These activities suggest that the recombinant CHIT42 acts as an endochitinase.Key words: Metarhizium anisopliae, chitinases, chit genes, recombinant protein, enthomopathogenic fungi.

scholarly journals Characterization of a new toxin from the entomopathogenic fungus Metarhizium anisopliae: the ribotoxin anisoplin

2017 ◽  
Vol 398 (1) ◽  
pp. 135-142 ◽  
Author(s):  
Miriam Olombrada ◽  
Pilar Medina ◽  
Flor Budia ◽  
José G. Gavilanes ◽  
Álvaro Martínez-del-Pozo ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Virulence Factors ◽  
Malaria Vector ◽  
Metarhizium Anisopliae ◽  
Entomopathogenic Fungus ◽  
Insect Cells ◽  
Malaria Vector Control ◽  
Ribonucleolytic Activity ◽  
Hirsutella Thompsonii

Abstract Metarhizium anisopliae is an entomopathogenic fungus relevant in biotechnology with applications like malaria vector control. Studies of its virulence factors are therefore of great interest. Fungal ribotoxins are toxic ribonucleases with extraordinary efficiency against ribosomes and suggested as potential insecticides. Here we describe this ribotoxin characteristic activity in M. anisopliae cultures. Anisoplin has been obtained as a recombinant protein and further characterized. It is structurally similar to hirsutellin A, the ribotoxin from the entomopathogen Hirsutella thompsonii. Moreover, anisoplin shows the ribonucleolytic activity typical of ribotoxins and cytotoxicity against insect cells. How Metarhizium uses this toxin and possible applications are of interest.

scholarly journals Characterization of a Dye-Decolorizing Peroxidase from Irpex lacteus Expressed in Escherichia coli: An Enzyme with Wide Substrate Specificity Able to Transform Lignosulfonates

2021 ◽  
Vol 7 (5) ◽  
pp. 325
Author(s):  
Laura Isabel de de Eugenio ◽  
Rosa Peces-Pérez ◽  
Dolores Linde ◽  
Alicia Prieto ◽  
Jorge Barriuso ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Inclusion Bodies ◽  
Veratryl Alcohol ◽  
Dye Decolorization ◽  
Irpex Lacteus ◽  
Type D ◽  
Lignin Peroxidases

A dye-decolorizing peroxidase (DyP) from Irpex lacteus was cloned and heterologously expressed as inclusion bodies in Escherichia coli. The protein was purified in one chromatographic step after its in vitro activation. It was active on ABTS, 2,6-dimethoxyphenol (DMP), and anthraquinoid and azo dyes as reported for other fungal DyPs, but it was also able to oxidize Mn2+ (as manganese peroxidases and versatile peroxidases) and veratryl alcohol (VA) (as lignin peroxidases and versatile peroxidases). This corroborated that I. lacteus DyPs are the only enzymes able to oxidize high redox potential dyes, VA and Mn+2. Phylogenetic analysis grouped this enzyme with other type D-DyPs from basidiomycetes. In addition to its interest for dye decolorization, the results of the transformation of softwood and hardwood lignosulfonates suggest a putative biological role of this enzyme in the degradation of phenolic lignin.

scholarly journals Characterization of Epidemic IncI1-Iγ Plasmids Harboring Ambler Class A and C Genes in Escherichia coli and Salmonella enterica from Animals and Humans

2015 ◽  
Vol 59 (9) ◽  
pp. 5357-5365 ◽  
Author(s):  
Hilde Smith ◽  
Alex Bossers ◽  
Frank Harders ◽  
Guanghui Wu ◽  
Neil Woodford ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enterica ◽  
Phylogenetic Trees ◽  
Functional Study ◽  
Content Type ◽  
Gene Associations ◽  
Genes Encoding ◽  
Marked Resistance

ABSTRACTThe aim of the study was to identify the plasmid-encoded factors contributing to the emergence and spread of epidemic IncI1-Iγ plasmids obtained fromEscherichia coliandSalmonella entericaisolates from animal and human reservoirs. For this, 251 IncI1-Iγ plasmids carrying various extended-spectrum β-lactamase (ESBL) or AmpC β-lactamase genes were compared using plasmid multilocus sequence typing (pMLST). Thirty-two of these plasmids belonging to different pMLST types were sequenced using Roche 454 and Illumina platforms. Epidemic IncI1-Iγ plasmids could be assigned to various dominant clades, whereas rarely detected plasmids clustered together as a distinct clade. Similar phylogenetic trees were obtained using only the plasmid backbone sequences, showing that the differences observed between the plasmids belonging to distinct clades resulted mainly from differences between their backbone sequences. Plasmids belonging to the various clades differed particularly in the presence/absence of genes encoding partitioning and addiction systems, which contribute to stable inheritance during cell division and plasmid maintenance. Despite this, plasmids belonging to the various phylogenetic clades also showed marked resistance gene associations, indicating the circulation of successful plasmid-gene combinations. The variation intraYandexcAgenes found in IncI1-Iγ plasmids is conserved within pMLST sequence types and plays a role in incompatibility, although functional study is needed to elucidate the role of these genes in plasmid epidemiology.

scholarly journals Molecular Characterization of Commensal Escherichia coli Adapted to Different Compartments of the Porcine Gastrointestinal Tract

2012 ◽  
Vol 78 (19) ◽  
pp. 6799-6803 ◽  
Author(s):  
Sam Abraham ◽  
David M. Gordon ◽  
James Chin ◽  
Huub J. M. Brouwers ◽  
Peter Njuguna ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gastrointestinal Tract ◽  
Random Amplified Polymorphic Dna ◽  
Content Type ◽  
E Coli ◽  
Plasmid Replicon ◽  
Different Strains ◽  
Different Characteristics

ABSTRACTThe role ofEscherichia colias a pathogen has been the focus of considerable study, while much less is known about it as a commensal and how it adapts to and colonizes different environmental niches within the mammalian gut. In this study, we characterizeEscherichia coliorganisms (n= 146) isolated from different regions of the intestinal tracts of eight pigs (dueodenum, ileum, colon, and feces). The isolates were typed using the method of random amplified polymorphic DNA (RAPD) and screened for the presence of bacteriocin genes and plasmid replicon types. Molecular analysis of variance using the RAPD data showed thatE. coliisolates are nonrandomly distributed among different gut regions, and that gut region accounted for 25% (P< 0.001) of the observed variation among strains. Bacteriocin screening revealed that a bacteriocin gene was detected in 45% of the isolates, with 43% carrying colicin genes and 3% carrying microcin genes. Of the bacteriocins observed (H47, E3, E1, E2, E7, Ia/Ib, and B/M), the frequency with which they were detected varied with respect to gut region for the colicins E2, E7, Ia/Ib, and B/M. The plasmid replicon typing gave rise to 25 profiles from the 13 Inc types detected. Inc F types were detected most frequently, followed by Inc HI1 and N types. Of the Inc types detected, 7 were nonrandomly distributed among isolates from the different regions of the gut. The results of this study indicate that not only may the different regions of the gastrointestinal tract harbor different strains ofE. colibut also that strains from different regions have different characteristics.

scholarly journals Cloning and expression of Saccharomyces cerevisiae copper-metallothionein gene in Escherichia coli and characterization of the recombinant protein

1993 ◽  
Vol 212 (2) ◽  
pp. 521-528 ◽  
Author(s):  
Zehra SAYERS ◽  
Patricia BROUILLON ◽  
Constantin E. VORGIAS ◽  
Hans F. NOLTING ◽  
Christoph HERMES ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Recombinant Protein ◽  
Cloning And Expression ◽  
Metallothionein Gene

scholarly journals Kinetic characterization of the Escherichia coli oligopeptidase A (OpdA) and the role of the Tyr607 residue

2010 ◽  
Vol 500 (2) ◽  
pp. 131-136 ◽  
Author(s):  
Ricardo Z. Lorenzon ◽  
Carlos E.L. Cunha ◽  
Marcelo F. Marcondes ◽  
Maurício F.M. Machado ◽  
Maria A. Juliano ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Kinetic Characterization

scholarly journals Cloning of the fabF gene in an expression vector and in vitro characterization of recombinant fabF and fabB encoded enzymes from Escherichia coli

FEBS Letters ◽  
1997 ◽  
Vol 402 (1) ◽  
pp. 62-66 ◽  
Author(s):  
Patricia Edwards ◽  
Janet Sabo Nelsen ◽  
James G Metz ◽  
Katayoon Dehesh
Keyword(s):  
Escherichia Coli ◽  
Expression Vector ◽  
In Vitro Characterization

scholarly journals Polyamine Transport and Role of potE in Response to Osmotic Stress in Escherichia coli

2000 ◽  
Vol 182 (21) ◽  
pp. 6247-6249 ◽  
Author(s):  
Dirk Schiller ◽  
Daniela Kruse ◽  
Helmut Kneifel ◽  
Reinhard Krämer ◽  
Andreas Burkovski
Keyword(s):  
Escherichia Coli ◽  
Osmotic Stress ◽  
Deletion Mutant ◽  
Transport Processes ◽  
Hyperosmotic Shock ◽  
Physiological Conditions ◽  
Polyamine Transport

ABSTRACT When transport of polyamines in Escherichia coli was examined, putrescine excretion was observed under two different physiological conditions: (i) strictly correlated to growth and (ii) following a hyperosmotic shock. Spermidine was not excreted. Characterization of a deletion mutant showed that PotE is not involved in these transport processes.

scholarly journals Characterization of the Induction and Cellular Role of the BaeSR Two-Component Envelope Stress Response of Escherichia coli

2011 ◽  
Vol 193 (13) ◽  
pp. 3367-3375 ◽  
Author(s):  
S. K. D. Leblanc ◽  
C. W. Oates ◽  
T. L. Raivio
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Envelope Stress ◽  
Two Component
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