Publications by authors named "Mikhail S Shupletsov"

Inorganic pyrophosphatases (PPases) catalyze an essential reaction, namely, the hydrolysis of PP, which is formed in large quantities as a side product of numerous cellular reactions. In the majority of living species, PP hydrolysis is carried out by soluble cytoplasmic PPase (S-PPases) with the released energy dissipated in the form of heat. In part of this energy can be conserved by proton-pumping pyrophosphatase (H-PPase) in the form of a proton electrochemical gradient for further ATP synthesis.

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Dynamic control is a distinguished strategy in modern metabolic engineering, in which inducible convergent transcription is an attractive approach for conditional gene silencing. Instead of a simple strong "reverse" () promoter, a three-component actuator has been developed for constitutive genes silencing. These actuators, consisting of promoters with different strengths, the ribosomal transcription antitermination-inducing sequence -AT, and the RNase III processing site, were inserted into the 3'-UTR of three metabolic genes.

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Background: Steady-state (13)C-based metabolic flux analysis ((13)C-MFA) is the most powerful method available for the quantification of intracellular fluxes. These analyses include concertedly linked experimental and computational stages: (i) assuming the metabolic model and optimizing the experimental design; (ii) feeding the investigated organism using a chosen (13)C-labeled substrate (tracer); (iii) measuring the extracellular effluxes and detecting the (13)C-patterns of intracellular metabolites; and (iv) computing flux parameters that minimize the differences between observed and simulated measurements, followed by evaluating flux statistics. In its early stages, (13)C-MFA was performed on the basis of data obtained in a single labeling experiment (SLE) followed by exploiting the developed high-performance computational software.

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