D-Proline reductases from Clostridium sticklandii and other species catalyze the reductive ring cleavage of D-proline to 5-aminovalerate. The reaction is physiologically significant in the anaerobic metabolism of many amino acids because it serves as an electron sink process and can be coupled to the oxidation of several different amino acids in a Stickland reaction. PrdA, the subunit of D-proline reductase from Clostridium sticklandii, is processed post-translationally to form a catalytic active pyruvoyl group. The peptide chain is cleaved into alpha and beta chains. The cleavage occurs on the N-terminal side of a cysteine residue which is the precursor of a pyruvoyl moiety. However, there is no successful reported case of in vitro post-translational modification of PrdA. In one recent study, it was found that a protein of the hdcAPB operon plays important role in post-translational cleavage of alpha subunit of pyruvoyl-dependent histidine decarboxylases from Streptococcus thermophilus CHCC1524 to form the pyruvoyl prosthetic group. In this study, we enhanced expression of recombinant prdA in Escherichia coli by codon optimization and expressed other unknown functional genes found in the prd operon with prdA to solve the expression and post-cleavage problems. The results of the experiment will be discussed in detail.

Expression and post-translational modification of recombinant D-Proline reductase from <i>Clostridium sticklandii</i> in <i>Escherichia coli</i>

247th National Meeting (2014)

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Expression and post-translational modification of recombinant D-Proline reductase from Clostridium sticklandii in Escherichia coli