title: Gene Expression of Lytic Endopeptidases AlpA and AlpB from Lysobacter sp. XL1 in Pseudomonads.
abstract: Development of an efficient expression system for (especially secreted) bacterial lytic enzymes is a complicated task due to the specificity of their action. The substrate for such enzymes is peptidoglycan, the main structural component of bacterial cell walls. For this reason, expression of recombinant lytic proteins is often accompanied with lysis of the producing bacterium. This paper presents data on the construction of an inducible system for expression of the lytic peptidases AlpA and AlpB from Lysobacter sp. XL1 in Pseudomonas fluorescens Q2-87, which provides for the successful secretion of these proteins into the culture liquid. In this system, the endopeptidase gene under control of the T7lac promoter was integrated into the bacterial chromosome, as well as the Escherichia coli lactose operon repressor protein gene. The T7 pol gene under lac promoter control, which encodes the phage T7 RNA polymerase, is maintained in Pseudomonas cells on the plasmids. Media and cultivation conditions for the recombinant strains were selected to enable the production of AlpA and AlpB by a simple purification protocol. Production of recombinant lytic enzymes should contribute to the development of new-generation antimicrobial drugs whose application will not be accompanied by selection of resistant microorganisms.
authors.affiliation: Laboratorie of Microbial Cell Surface Biochemistry, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Russia.
authors.forename: Irina M
authors.initials: IM
authors.lastname: Tsfasman
authors.forename: Yulia S
authors.initials: YS
authors.lastname: Lapteva
authors.forename: Ludmila A
authors.initials: LA
authors.lastname: Krasovskaya
authors.forename: Irina V
authors.initials: IV
authors.lastname: Kudryakova
authors.forename: Natalia V
authors.initials: NV
authors.lastname: Vasilyeva
authors.forename: Igor E
authors.initials: IE
authors.lastname: Granovsky
authors.forename: Olga A
authors.initials: OA
authors.lastname: Stepnaya
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism; D020563:Gammaproteobacteria / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D053528:Lysobacter / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D010450:Endopeptidases