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

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 (1985-04-01)
P Nedkov, W Oberthür, G 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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Proteinase, bacterial, Type XXIV, 7.0-14.0 units/mg solid, lyophilized powder
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Protease from Bacillus licheniformis, glycerol solution (50%)
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Protease from Bacillus licheniformis, Type VIII, lyophilized powder, 7-15 units/mg solid
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Protease from Bacillus licheniformis, lyophilized powder, for use in Total Dietary Fiber Assay, TDF-100A
Sigma-Aldrich
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