Lytic transglycosylase MltB of Escherichia coli and its role in recycling of peptidoglycan strands of bacterial cell wall.
The cell wall is an indispensable structure for the survival of bacteria and a target for antibiotics. Peptidoglycan is the major constituent of the cell wall, which is comprised of backbone repeats of N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM). A peptide stem is appended to the NAM unit, which in turn experiences cross-linking with a peptide from another peptidoglycan in the final steps of cell wall assembly. In the normal course of bacterial growth, as much as 60% of the parental cell wall is recycled, a process that is not fully understood. A polymeric cell wall is fragmented by the family of lytic transglycosylases, and certain key fragments are transported to the cytoplasm for recycling. The genes for the six known lytic transglycosylases of Escherichia coli were cloned, and the enzymes were purified in this study. It is shown that MltB is the only lytic transglycosylase to turn over a synthetic peptidoglycan fragment of two NAG-NAM repeats; hence this enzyme is likely to be the lytic transglycosylase responsible for processing of shorter peptidoglycan strands. Lytic transglycosylases have been proposed to go through an oxocarbenium species that would trap the 6-hydroxyl moiety of the glucosamine residue of muramic acid to generate the so-called 1,6-anhydromuramyl moiety. It is documented herein by characterization of the products of turnover that this process takes place to the total exclusion of the entrapment of a water molecule by the reactive intermediary oxocarbenium species. Furthermore, turnover of the E. coli sacculus (whole cell wall) by MltB was characterized. It is documented that each MltB molecule is able to process the cell wall 14000 times in the course of a single doubling time for E. coli.
Suvorov
Maxim
M
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, USA.
Lee
Mijoon
M
Hesek
Dusan
D
Boggess
Bill
B
Mobashery
Shahriar
S
D002473:Cell Wall / Q000201:enzymology / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016695:Glycosyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010457:Peptidoglycan / Q000378:metabolism*
D029968:Escherichia coli Proteins; D010457:Peptidoglycan; D016695:Glycosyltransferases; C021815:murein transglycosylase