Journal ArticleDOI
Overexpression of Escherichia coli genes encoding nucleoside phosphorylases in the pET/Bl21(DE3) system yields active recombinant enzymes.
Roman S. Esipov,Gurevich Ai,Dmitry V. Chuvikovsky,L. A. Chupova,Tatyana I. Muravyova,Anatoly I. Miroshnikov +5 more
TLDR
Optimum conditions for biosynthesis of each enzyme as a soluble protein with intact biological activity were found and the crude preparations are approximately 80% pure and can be used immediately for enzymatic transglycosylation.About:
This article is published in Protein Expression and Purification.The article was published on 2002-02-01. It has received 53 citations till now. The article focuses on the topics: Purine nucleoside phosphorylase & Uridine phosphorylase.read more
Citations
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Journal ArticleDOI
Biologically important nucleosides: modern trends in biotechnology and application☆
TL;DR: Some aspects of the functioning of enzymes of nucleic acid metabolism of interest as biocatalysts, the transglycosylation reaction using sugar modified nucleosides as donors of carbohydrate residues and heterocyclic bases as acceptors catalyzed by nucleoside phosphorylases and N -deoxyribosyltransferases, and the retrosynthesis are considered.
Journal ArticleDOI
Immobilized Biocatalysts for the Production of Nucleosides and Nucleoside Analogues by Enzymatic Transglycosylation Reactions
Gabriele Zuffi,Daniela Ghisotti,Ilaria Oliva,Emanuele Capra,Gianni Frascotti,Giancarlo Tonon,Gaetano Orsini +6 more
TL;DR: The new biocatalysts described in this work are suitable for both laboratory and industrial scale applications due to the maintainance of high catalytic efficiency, thermal and solvent stability, reusability and ease of operation in batch as well as in continuous reactions.
Journal ArticleDOI
Validation of the catalytic mechanism of Escherichia coli purine nucleoside phosphorylase by structural and kinetic studies
Goran Mikleušević,Zoran Štefanić,Marta Narczyk,Beata Wielgus-Kutrowska,Agnieszka Bzowska,Marija Luić +5 more
TL;DR: The catalytic mechanism of Escherichia coli purine nucleoside phosphorylase is revised using site-directed mutagenesis, kinetic studies and structure determinations, which show that previously observed conformational change is a result of the phosphate binding and its interaction with Arg24.
Journal ArticleDOI
The chemoenzymatic synthesis of clofarabine and related 2′-deoxyfluoroarabinosyl nucleosides: the electronic and stereochemical factors determining substrate recognition by E. coli nucleoside phosphorylases
Ilja V. Fateev,Konstantin V. Antonov,Irina D. Konstantinova,Tatyana I. Muravyova,Frank Seela,Roman S. Esipov,Anatoly I. Miroshnikov,Igor A. Mikhailopulo +7 more
TL;DR: Two approaches to the synthesis of 2-chloro-9-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)adenine (1, clofarabine) were studied and differences of the reactivity of various substrates were analyzed by ab initio calculations.
Journal ArticleDOI
Enzymatic transglycosylation of natural and modified nucleosides by immobilized thermostable nucleoside phosphorylases from Geobacillus stearothermophilus
TL;DR: Owing to the high catalytic activity, thermal stability, the ease of application, and the possibility of repeated use, the immobilized preparations of thermostable nucleoside phosphorylases are suitable for the production of pharmacologically important natural and modified nucleosides.
References
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Journal ArticleDOI
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Journal ArticleDOI
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal Article
Cleavage of structural proteins during the assemble of the head of bacterio-phage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.