Open AccessTwo‐step procedure for purification and separation of the essential penicillin‐binding proteins PBP 1A and 1Bs of Escherichia coli
Author(s) -
Rechenberg Moritz,
Höltje JoachimVolker
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09230.x
Subject(s) - escherichia coli , biochemistry , penicillin binding proteins , biology , polymerase , enzyme , bifunctional , gene , catalysis
The penicillin‐binding proteins PBP 1A and 1Bs are the essential murein polymerases of Escherichia coli . Purification of these membrane‐bound bifunctional transglycosylase‐transpeptidases was a major obstacle in studying the details of both enzymatic reactions. Here we describe a simple, highly specific affinity chromatography method that takes advantage of the availability of the specific inhibitor of the transglycosylase site moenomycin A in order to enrich PBP 1A and 1Bs in one step from crude membrane preparations. Separation of PBP 1A from PBP 1Bs is achieved in a second step employing cation exchange chromatography yielding enzymatically active native murein polymerases.