Biotechnological potential of microbial α-galactosidases.

The enzyme α-galactosidase (α-D-galactoside galactohydrolase; EC 3.2.1.

Propionate sensor using coenzyme-A transferase and acyl-CoA oxidase.

We developed an amperometric propionate sensor using comprised of two recombinant enzymes, propionate coenzyme A CoA transferase from Clostridium propionicum and short-chain acyl-CoA oxidase from Arabidopsis thaliana. Response current increased linearly with increase in propionate concentration from 10 microM to 100 microM. The detection limit was 10 microM propionate.

MmcBC in Pelotomaculum thermopropionicum represents a novel group of prokaryotic fumarases.

The overall amino-acid sequence of MmcBC in Pelotomaculum thermopropionicum was substantially homologous (33%) to fumarase A in Escherichia coli, although its possible subunit structure was different from known fumarases and it lacked the fumarate-lyase signature sequence. Here, MmcBC in E. coli is expressed and characterized.

Volatile fatty acid-sensing system involving coenzyme-A transferase.

On-site monitoring of volatile fatty acids (VFAs), such as propionate, is industrially and medically important. The present study developed a VFA biosensing system comprised of two recombinant enzymes, propionate coenzyme A (CoA) transferase (PCT) from Clostridium propionicum and acyl-CoA oxidase from Arabidopsis thaliana. This system produced hydrogen peroxide in the presence of acetyl-CoA, oxygen, and VFA substrates, which could be quantified by colorimetric methods using peroxidase and dye reagents (e.

Propionyl-coenzyme A synthetases of Ralstonia solanacearum and Salmonella choleraesuis display atypical kinetics.

Propionyl-coenzyme A synthetases (PrpE) of Salmonella choleraesuis and Ralstonia solanacearum sharing 62% identity in amino acid sequence to each other were cloned, expressed in Escherichia coli and purified. Both enzymes catalyzed acetyl-, propionyl-, butyryl- and acrylyl-coenzyme A formation with the highest k(cat)/K(m) values for propionate. They displayed sigmoidal homotrophic autoactivation kinetics for propionate but not for the other acyl substrates tested.

Characterization of a cellobiose phosphorylase from a hyperthermophilic eubacterium, Thermotoga maritima MSB8.

The cepA putative gene encoding a cellobiose phosphorylase of Thermotoga maritima MSB8 was cloned, expressed in Escherichia coli BL21-codonplus-RIL and characterized in detail. The maximal enzyme activity was observed at pH 6.2 and 80 degrees C.