Preparation and characterization of recombinant protein ScFv(CD11c)-TRP2 for tumor therapy from inclusion bodies in Escherichia coli

2007 ◽  
Vol 52 (1) ◽  
pp. 131-138 ◽  
Author(s):  
Geng Kou ◽  
Shu Shi ◽  
Hao Wang ◽  
Min Tan ◽  
Jingya Xue ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Inclusion Bodies ◽  
Tumor Therapy

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.

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 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

Isolation and partial characterization of the Asp-B10, Tyr-B25-des-(B26-B30)-proinsulin analog from inclusion bodies in Escherichia coli

1996 ◽  
Vol 10 (10) ◽  
Author(s):  
A.M.A. Nascimento ◽  
H.R.T. Souza ◽  
M.S. Xavier ◽  
J.E. Thiemann ◽  
L. Vilela ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Inclusion Bodies ◽  
Partial Characterization

Expression of a Brassic napus glutamate 1-semialdehyde aminotransferase in Escherichia coli and characterization of the recombinant protein

2003 ◽  
Vol 29 (2) ◽  
pp. 193-201 ◽  
Author(s):  
Edward W.T Tsang ◽  
Zhiyuan Hu ◽  
Qing Chang ◽  
D.Ian McGregor ◽  
Wilfred A Keller
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein

scholarly journals Production and Characterization of ZFP36L1 Antiserum against Recombinant Protein from Escherichia coli

2008 ◽  
Vol 24 (2) ◽  
pp. 326-333 ◽  
Author(s):  
H. Cao ◽  
R. Lin ◽  
S. Ghosh ◽  
R.A. Anderson ◽  
J.F. Urban
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein

scholarly journals Nucleic Acids in Inclusion Bodies obtained from E. coli Cells Expressing Human Interferon-Gamma

2020 ◽  
Author(s):  
Elena Krachmarova ◽  
Ivan Ivanov ◽  
Genoveva Nacheva
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Nucleic Acids ◽  
Interferon Gamma ◽  
Recombinant Proteins ◽  
Gel Electrophoresis ◽  
Inclusion Bodies ◽  
Protein Aggregates ◽  
Human Interferon ◽  
Molecular Heterogeneity

Abstract Background Inclusion bodies (IBs) are protein aggregates in recombinant bacterial cells containing mainly the target recombinant protein. Although it has been shown that IBs contain functional proteins along with protein aggregates, their direct application as pharmaceuticals is hindered by their heterogeneity and hazardous contaminants with bacterial origin. Therefore, together with the production of soluble species, IBs remain the main source for manufacture of recombinant proteins with medical application. The quality and composition of the IBs affect the refolding yield and further purification of the recombinant protein. The knowledge whether nucleic acids are genuine components or concomitant impurities of the IBs is a prerequisite for the understanding of the IBs formation and for development of optimized protocols for recombinant protein refolding and purification. IBs isolated from Escherichia coli overexpressing human interferon-gamma (hIFNγ), a protein with therapeutic application, were used as a model. Results IBs were isolated from Escherichia coli LE392 cells transformed with a hIFNγ expressing plasmid under standard conditions and further purified by centrifugation on a sucrose cushion, followed by several steps of sonication and washings with non-denaturing concentrations of urea. The efficiency of the purification was estimated by SDS-PAGE gel electrophoresis and parallel microbiological testing for the presence of residual intact bacteria. Phenol/chloroform extraction showed that the highly purified IBs contain both DNA and RNA. The latter were studied by UV spectroscopy and agarose gel electrophoresis combined with enzymatic treatment and hybridization. DNA was observed as a diffuse fraction mainly in the range of 250 to 1000 bp. RNA isolated by TRIzol® also demonstrated a substantial molecular heterogeneity. Hybridization with 32 P-labelled oligonucleotides showed that the IBs contain rRNA and are enriched of hIFNγ mRNA. Conclusions The results presented in this study indicate that the nucleic acids are intrinsic components rather than co-precipitated impurities in the IBs. We assume that the nucleic acids are active participants in the aggregation of recombinant proteins and formation of the IBs that originate from the transcription and translation machinery of the microbial cell factory.

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