Showing papers by "Anatoly I. Miroshnikov published in 2008"
TL;DR: It was necessary to work out optimal scheme for refolding and purification of the recombinant hEGF, a hybrid protein that had several cysteins in its sequence and was capable under certain conditions of autocatalytic cleavage resulting in separation of the target protein.
40 citations
TL;DR: Cladribine (2-chloro-2′-deoxyadenosine) was synthesized using intact cells of the recombinant Escherichia coli strain producing Geobacillus stearothermophilus B-2194 thermostable purine-nucleoside phosphorylase II (EC 2.4.2.1), which provided the maximal concentrations of sparingly soluble substrates.
Abstract: Cladribine (2-chloro-2′-deoxyadenosine) was synthesized using intact cells of the recombinant Escherichia coli strain producing Geobacillus stearothermophilus B-2194 thermostable purine-nucleoside phosphorylase II (EC 2.4.2.1). Use of the cells containing this thermostable enzyme allowed the process to be conducted at a temperature of 70°C, which provided the maximal concentrations of sparingly soluble substrates. The best results were obtained with 2-chloroadenine as a modified base. The highest yield of the target 2-chloro-2′-deoxyadenosine (up to 95% in the case of deoxyguanosine) was reached when using 2′-deoxypurines as donors of deoxyribose. Use of thymidine for these purposes required its considerable molar excess over 2-chloroadenine (up to 6:1), which is connected with a nonoptimal amount of endogenous thymidine phosphorylase, necessary for synthesis of deoxyribose-1-phosphate, in the transglycosylation reaction.
13 citations
01 Sep 2008
8 citations
TL;DR: The stable strain of methylotrophic yeast Pichia pastoris secreting human serum albumin into cultural medium was obtained and recombinant protein was characterized by mass spectrometry and circular dichroism and analyzed its catalytic activity.
Abstract: The stable strain of methylotrophic yeast Pichia pastoris secreting human serum albumin into cultural medium was obtained. Optimal conditions for expression of the protein were determined. We characterized the recombinant protein by mass spectrometry and circular dichroism and analyzed its catalytic activity.
3 citations
TL;DR: In this paper, an efficient enzymatic method is suggested for the synthesis of purine nucleosides containing diverse substituents at the C6 and C2 carbon atoms.
Abstract: An enzymatic transglycosylation of purine heterocyclic bases employing readily available natural nucleosides or sugar-modified nucleosides as donors of the pentofuranose fragment and recombinant nucleoside phosphorylases as biocatalysts has been investigated. An efficient enzymatic method is suggested for the synthesis of purine nucleosides containing diverse substituents at the C6 and C2 carbon atoms. The glycosylation of N6-benzoyladenine and N2-acetylguanine and its O6-derivatives is not accompanied by deacylation of bases.
3 citations