title: Proteolytic modification of Leuconostoc mesenteroides B-512F dextransucrase.
abstract: Multiple active lower molecular weight forms from Leuconostoc mesenteroides B512F dextransucrase have been reported. It has been suggested that they arise from proteolytic processing of a 170 kDa precursor. In this work, the simultaneous production of proteases and dextransucrase was studied in order to elucidate the dextransucrase proteolytic processing. The effect of the nitrogen source on protease and dextransucrase production was studied. Protease activity reaches a maximum early in the logarithmic phase of dextransucrase synthesis using the basal culture medium but the nitrogen source plays an important effect on growth: the highest protease concentration was obtained when ammonium sulfate, casaminoacids or tryptone were used. Two active forms of 155 and 129 kDa were systematically obtained from dextransucrase precursor by proteolysis. The amino termini of these forms were sequenced and the cleavage site deduced. Both forms of the enzyme obtained had the same cleavage site in the amino terminal region (F209-Y210). From dextransucrase analysis, various putative cleavage sites with the same sequence were found in the variable region and in the glucan binding domain. Although no structural differences were found in dextrans synthesized with both the precursor and the proteolyzed 155 kDa form under the same reaction conditions, their rheological behaviour was modified, with dextran of a lower viscosity yielded by the smaller form.
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, 62250, Morelos, Cuernavaca, México.
authors.forename: Martha
authors.initials: M
authors.lastname: Argüello-Morales
authors.forename: Mónica
authors.initials: M
authors.lastname: Sánchez-González
authors.forename: Mariana
authors.initials: M
authors.lastname: Canedo
authors.forename: Maricarmen
authors.initials: M
authors.lastname: Quirasco
authors.forename: Amélia
authors.initials: A
authors.lastname: Farrés
authors.forename: Agustín
authors.initials: A
authors.lastname: López-Munguía
mesh_terms: D003911:Dextrans / Q000096:biosynthesis / Q000737:chemistry; D005964:Glucosyltransferases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D007934:Leuconostoc / Q000201:enzymology*; D008970:Molecular Weight; D009844:Oligosaccharides / Q000096:biosynthesis / Q000737:chemistry; D010447:Peptide Hydrolases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D020539:Sequence Analysis, Protein
chemical_list: D003911:Dextrans; D009844:Oligosaccharides; D005964:Glucosyltransferases; C020025:dextransucrase; D010447:Peptide Hydrolases