By a News Reporter-Staff News Editor at Biotech Week -- Fresh data on Enzymes and Coenzymes are presented in a new report. According to news reporting out of Jeddah, Saudi Arabia, by NewsRx editors, research stated, "In this study, we have investigated a propionate CoA-transferase (Pct) homologue encoded in the genome of Ralstonia eutropha H16. The corresponding gene has been cloned into the vector pET-19b to yield a histidine-tagged enzyme which was expressed in Escherichia coli BL21 (DE3)."
Our news journalists obtained a quote from the research from King Abdul-Aziz University, "After purification, high-performance liquid chromatography/mass spectrometry (HPLC/MS) analyses revealed that the enzyme exhibits a broad substrate specificity for carboxylic acids. The formation of the corresponding CoA-thioesters of acetate using propionyl-CoA as CoA donor, and of propionate, butyrate, 3-hydroxybutyrate, 3-hydroxypropionate, crotonate, acrylate, lactate, succinate and 4-hydroxybutyrate using acetyl-CoA as CoA donor could be shown. According to the substrate specificity, the enzyme can be allocated in the family I of CoA-transferases. The apparent molecular masses as determined by gel filtration and detected by SDS polyacrylamide gel electrophoresis were 228 and 64 kDa, respectively, and point to a quaternary structure of the native enzyme (alpha(4)). The enzyme exhibited similarities in sequence and structure to the well investigated Pct of Clostridium propionicum. It does not contain the typical conserved (S)ENG motif, but the derived motif sequence EXG with glutamate 342 to be, most likely, the catalytic residue. Due to the homo-oligomeric structure and the sequence differences with the subclasses IA-C of family I CoA-transferases, a fourth subclass of family I is proposed, comprising - amongst others - the Pcts of R. eutropha H16 and C. propionicum. A markerless precise-deletion mutant R. eutropha H16a dagger pct was generated. The growth and accumulation behaviour of this mutant on gluconate, gluconate plus 3,3'-dithiodipropionic acid (DTDP), acetate and propionate was investigated but resulted in no observable phenotype. Both, the wild type and the mutant showed the same growth and storage behaviour with these carbon sources. It is probable that R. eutropha H16 is upregulating other CoA-transferase(s) or CoA-synthetase(s), thereby compensating for the lacking Pct."
According to the news editors, the research concluded: "The ability of R. eutropha H16 to substitute absent enzymes by isoenzymes has been already shown in different other studies in the past."
For more information on this research see: A propionate CoA-transferase of Ralstonia eutropha H16 with broad substrate specificity catalyzing the CoA thioester formation of various carboxylic acids. Applied Microbiology and Biotechnology, 2013;97(17):7699-7709. Applied Microbiology and Biotechnology can be contacted at: Springer, 233 Spring St, New York, NY 10013, USA. (Springer - www.springer.com; Applied Microbiology and Biotechnology - www.springerlink.com/content/0175-7598/)
Our news journalists report that additional information may be obtained by contacting N. Lindenkamp, King Abdulaziz Univ, Dept. of Environm Sci, Jeddah 21413, Saudi Arabia. Additional authors for this research include M. Schurmann and A. Steinbuchel (see also Enzymes and Coenzymes).
Keywords for this news article include: Asia, Jeddah, Propionates, Saudi Arabia, Transferases, Acyclic Acids, Carboxylic Acids, Organic Chemicals, Enzymes and Coenzymes
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