A 4-amino-2-hydroxybutyrate activating enzyme from butirosin-producing Bacillus circulans.
An enzyme catalyzing a 4-amino-2-hydroxybutyrate dependent ATP-PP(i)-exchange reaction has been partially purified from butirosin-producing cells of B. circulans by ammonium sulfate fractionation, gel filtration or sucrose gradient centrifugation and ion exchange chromatography on QAE-sepharose. No reaction was found with 4-aminobutyrate and diaminobutyrate. The protein coeluted with GroEL, which has been identified by alignment of the internal sequence KDGVITVEESK. A molecular mass of about 1,5 MDa has been estimated for the complex by gradient centrifugation. In SDS-polyacrylamide gels a protein band with an estimated size of about 500 KDa correlated with the enzyme activity. The activity was not found in non-producing cells and a non-producing mutant of B. circulans with an insertional inactivation in the butB gene.
Aubert-Pivert
E
E
Institut Pasteur, Unité de Génétique Mycobactérienne, Paris.
el-Samaraie
A
A
von Döhren
H
H
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D002076:Butirosin Sulfate / Q000096:biosynthesis*; D002499:Centrifugation, Density Gradient; D018834:Chaperonin 60; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008970:Molecular Weight; D016254:Mutagenesis, Insertional; D009713:Nucleotidyltransferases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D001426:Bacterial Proteins; D018834:Chaperonin 60; D006360:Heat-Shock Proteins; D002076:Butirosin Sulfate; D009713:Nucleotidyltransferases