title: Glutamyl endopeptidase of Bacillus intermedius strain 3-19. Purification, properties, and crystallization.
abstract: A homogeneous glutamyl endopeptidase splitting peptide bonds of glutamic and, rarely, of aspartic acid residues in peptides and proteins was isolated from Bacillus intermedius 3-19 culture filtrate using chromatography on CM-cellulose and Mono S. The enzyme molecular mass is 29 kD and the pI is 8.4. The proteinase is inhibited by DFP. The enzyme, like other glutamyl endopeptidases, reveals two pH optima (pH 7.5 and 9.0) for casein and one (pH 8.0) for Z-Glu-pNA hydrolysis. The K(m) for the hydrolysis of the latter substrate is 6 mM. The enzyme activity is optimal at 55 degrees C. The enzyme is stable in the pH range 6.5-11.0. Its N-terminal sequence shows 56% coinciding residues when compared with that of Bacillus licheniformis glutamyl endopeptidase. Crystal prisms or plates 0.25-0.3 x 0.15 x 0.07-0.1 mm have been grown using the vapor diffusion technique in a hanging drop followed by macroseeding. The crystals belong to the space group B2 with the following unit cell parameters: a = 69.59 A; b = 61.61 A; c = 56.11 A; gamma = 117.57 degrees. The X-ray data set to 1.7 A resolution has been collected on an automatic synchrotron (EMBL Hamburg Station).
authors.affiliation: Department of Microbiology, School of Biology, Kazan State University, Russia. root@scikgu.kazan.su
authors.forename: I B
authors.initials: IB
authors.lastname: Leshchinskaya
authors.forename: E V
authors.initials: EV
authors.lastname: Shakirov
authors.forename: E L
authors.initials: EL
authors.lastname: Itskovitch
authors.forename: N P
authors.initials: NP
authors.lastname: Balaban
authors.forename: A M
authors.initials: AM
authors.lastname: Mardanova
authors.forename: M R
authors.initials: MR
authors.lastname: Sharipova
authors.forename: E V
authors.initials: EV
authors.lastname: Blagova
authors.forename: V M
authors.initials: VM
authors.lastname: Levdikov
authors.forename: I P
authors.initials: IP
authors.lastname: Kuranova
authors.forename: G N
authors.initials: GN
authors.lastname: Rudenskaya
authors.forename: V M
authors.initials: VM
authors.lastname: Stepanov
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology*; D003460:Crystallization; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D011480:Protease Inhibitors; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase