CHEMICAL SYNTHESIS AND EXPRESSION OF A GENE CODING FOR HUMAN MUSCLE ACYLPHOSPHATASE

A DNA sequence coding for human muscle acylphosphatase has been constr ucted using 16 chemically synthesized oligonucleotides. The 300-bases long DNA sequence has been cloned in the pT7.7 Escherichia coli expres sion vector and in the pYEpsec1 Saccharomyces cerevisiae expression ve ctor. In both cases a high level of expression of acylphosphatase has been observed. The recombinant proteins have been purified to homogene ity and assayed in comparison with the natural protein, using benzoylp hosphate as a substrate and phosphate as a competitive inhibitor. The recombinant enzymes expressed in the two microorganisms maintain the k inetic properties of the natural protein. In addition, NMR analysis sh ows that the gross fold of the two recombinant enzymes is correct.