Mutagenic Characterization of a Conserved Functional Amino Acid in Fuculose-1-Phosphate Aldolase from Methanococcus jannaschii, a Hyperthermophic Archaea

Mutagenic Characterization of a Conserved Functional Amino Acid in Fuculose-1-Phosphate Aldolase from Methanococcus jannaschii, a Hyperthermophic Archaea

To elucidate the putative role of the amido group in the metal binding of the fuculose-1-phosphate aldolase from Methanococcus jannaschii, we have examined a potential targen using site-directed mutagenesis. The replacement of asparagine 25 with leucine or threonine was shown to have a negative effect, not only on catlytic efficiency, but also on substrage recognition as well. The Hill coefficient values yeilded a value of =1. All metals used with the wild-type aldolases exhibited higher activity than that of the mutants. The spectra of the mutants were quite different from the wild-type aldolase. A highly conserved amino acid of asparagine 25 in a related family of aldolase odes not appear to provide sufficient evidence for evolution.