Using the DNA recombination technique, a new bacterial strain Escherichia coli DAC-22 was derived, whose cells are able to carry out heterologous expression of Bacillus thuringiensis diadenylate cyclase – enzyme catalyzing reaction adenosine-5′-triphosphate (ATP) transformation into cyclic 3′,5′-diadenylate (cyclo-di-AMP). To derive strain, E. “Rosetta (DE3) pLysS” were originally used as recip...

A gene encoding a putative 150-amino-acid methylglyoxal synthase was identified in Clostridium acetobutylicum ATCC 824. The enzyme was overexpressed in Escherichia coli and purified. Methylglyoxal synthase has a native molecular mass of 60 kDa and an optimum pH of 7.5. The Km and Vmax values for the substrate dihydroxyacetone phosphate were 0.53 mM and 1.56 mmol min(-1) microgram(-1), respectiv...

Here we describe a half-semester biochemistry laboratory course based on the expression, isolation, purification, and biochemical characterization of RecJ nuclease. The students transformed expression plasmid thermostable nuclease into host bacteria Escherichia coli BL21(DE3), to induce recombinant by IPTG. expressed was purified from cell lysates broken with sonication Ni-affinity chromatograp...

BACKGROUND Pseudomonas putida (P. putida) ATCC12633 can produce creatinase. It is a microbial enzyme which degrades creatinine in bacteria and provides source of carbon and nitrogen. Also, this enzyme is used in the enzymatic measurement of creatinine concentration for diagnosis of renal and muscles functions and diseases. Our purpose was recombinant production of creatinase for using in clinic...

DPPD (Rv0061) is a difficult to express protein of Mycobacterium tuberculosis that elicits strong and specific delayed type hypersensitivity reactions in humans infected with M. tuberculosis. Therefore e DPPD is a molecule that can improve the specificity of the tuberculin skin test, which is widely used as an aid for the diagnosis of tuberculosis. However, a pitfall of our initial studies was ...

Background: Some resources have suggested that genetically inactivated pertussis toxoid (PTs) bear a more protective effect than chemically inactivated products. This study aimed to produce new version of PT, by cloning an inactive pertussis toxin S1 subunit (PTS1) in a fusion form with N-terminal half of the listeriolysin O (LLO) pore-forming toxin. Methods: Deposited pdb structure file of the...

E. coli O157:H7, one of the major EHEC serotypes, is capable of developing bloody diarrhea, hemorrhagic colitis (HC), and fatal hemolytic uremic syndrome (HUS) and is accompanied by high annual economic loss worldwide. Due to the increased risk of HC and HUS development following antibiotic therapy, the prevention of infections caused by this pathogen is considered to be one of the most effecti...

conclusions: our data showed that the recombinant mature lysostaphin protein produced by pet32a vector in e. coli system was very efficient. results: pcr and sequencing results confirmed the successful cloning of the target gene into the vector. the expression of protein was induced by iptg and high concentration of the recombinant protein was obtained via the purification process by affinity-c...