title: [Cloning and expression of variants of the glutaryl-7-aminocephalosporic acid acylase of the bacterium Brevundimonas diminuta in Escherichia coli cells].
abstract: The gene coding for glutaryl-7-aminocephalosporic acid acylase (Gl7ACA acylase) of the bacterium Brevundimonas diminuta (BrdGl7ACA), a commercial enzyme widely used in modem biocatalytic technologies for manufacture of b-lactam antibiotics, was cloned. Efficient expression systems for producing a "native" recombinant BrdGl7ACA and its analogs modified by attaching affinity groups--the chitin-binding domain of chitinases A1 and hexahistidine sequence--were designed. It was demonstrated that both the recombinant hybrid proteins and the native Gl7ACA acylase produced in E. coli cells underwent a correct autoproteolytic processing with generation of functionally active enzymes and could be isolated with a high yield using one-step affinity chromatography.
authors.forename: S A
authors.initials: SA
authors.lastname: Khatuntseva
authors.forename: M A
authors.initials: MA
authors.lastname: El'darov
authors.forename: S A
authors.initials: SA
authors.lastname: Lopatin
authors.forename: O A
authors.initials: OA
authors.lastname: Zeĭnalov
authors.forename: K G
authors.initials: KG
authors.lastname: Skriabin
mesh_terms: D042063:Caulobacteraceae / Q000201:enzymology*; D002686:Chitin; D003001:Cloning, Molecular; D004800:Enzymes, Immobilized; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D010399:Penicillin Amidase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004800:Enzymes, Immobilized; D007527:Isoenzymes; D011994:Recombinant Proteins; D002686:Chitin; D010399:Penicillin Amidase; C046586:glutarylamidocephalosporanic acid acylase