Determination of the complete amino-acid sequence of subtilisin DY and its comparison with the primary structures of the subtilisins BPN', Carlsberg and amylosacchariticus

Biological Chemistry Hoppe-Seyler
P NedkovG Braunitzer

Abstract

The complete amino-acid sequence of subtilisin DY, an extracellular alkaline proteinase produced by Bacillus subtilis strain DY was determined. This included automated sequence analysis of the whole molecule and its large fragments such as tryptic peptides obtained from the inactivated enzyme, peptides generated by cyanogen bromide, by o-iodosobenzoic acid and by hydroxylamine. The peptides were isolated by gel filtration and by reversed-phase high performance liquid chromatography. The amino-acid sequence of subtilisin DY was determined by overlapping the isolated peptides. It consists of 274 amino-acid residues, like that of subtilisin Carlsberg. By comparison with the structures of the subtilisins Carlsberg, amylosacchariticus and BPN' 32, 80 and 82 amino-acid substitutions were found, which are caused by 37, 102 and 106 nucleotide mutations, respectively. It was found also that 62.5% of the amino-acid residues in the molecules of these four subtilisins are identical with respect to kind and position of the residue, which suggests that these molecules have had a common ancestral precursor. The amino-acid replacement analysis of the four subtilisins leads to the conclusion that they have evolved almost independently.

References

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Citations

Apr 1, 1995·Current Microbiology·Y YamagataE Ichishima
Mar 1, 1968·Proceedings of the National Academy of Sciences of the United States of America·A N Glazer
Dec 1, 1968·Proceedings of the National Academy of Sciences of the United States of America·F S MarklandE L Smith
Sep 1, 1986·Biological Chemistry Hoppe-Seyler·A LilovaG Braunitzer
Nov 1, 1987·Biological Chemistry Hoppe-Seyler·A LilovaP Nedkov
May 1, 1987·Biological Chemistry Hoppe-Seyler·A LilovaP Nedkov
Jan 1, 1988·Critical Reviews in Biotechnology·M L RollenceP N Bryan
Apr 15, 1992·Biochemical and Biophysical Research Communications·J S JangS M Byun
Mar 17, 2019·International Journal of Molecular Sciences·Raquel Santos-de-SouzaFranklin Souza-Silva
Aug 24, 2000·Journal of Agricultural and Food Chemistry·C T ChangH Y Sung

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