The periplasmic cyclodextrin binding protein CymE from Klebsiella oxytoca and its role in maltodextrin and cyclodextrin transport.
Klebsiella oxytoca M5a1 has the capacity to transport and to metabolize alpha-, beta- and gamma-cyclodextrins. Cyclodextrin transport is mediated by the products of the cymE, cymF, cymG, cymD, and cymA genes, which are functionally homologous to the malE, malF, malG, malK, and lamB gene products of Escherichia coli. CymE, which is the periplasmic binding protein, has been overproduced and purified. By substrate-induced fluorescence quenching, the binding of ligands was analyzed. CymE bound alpha-cyclodextrin, beta-cyclodextrin, and gamma-cyclodextrin, with dissociation constants (Kd) of 0.02, 0.14 and 0.30 microM, respectively, and linear maltoheptaose, with a Kd of 70 microM. In transport experiments, alpha-cyclodextrin was taken up by the cym system of K. oxytoca three to five times less efficiently than maltohexaose by the E. coli maltose system. Besides alpha-cyclodextrin, maltohexaose was also taken up by the K. oxytoca cym system, but because of the inability of maltodextrins to induce the cym system, growth of E. coli mal mutants on linear maltodextrin was not observed when the cells harbored only the cym uptake system. Strains which gained this capacity by mutation could easily be selected, however.
Pajatsch
M
M
Lehrstuhl für Mikrobiologie der Universität München, D-80638 München, Germany.
Gerhart
M
M
Peist
R
R
Horlacher
R
R
Boos
W
W
Böck
A
A
D001426:Bacterial Proteins / Q000378:metabolism*; D001692:Biological Transport; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D003505:Cyclodextrins / Q000378:metabolism*; D005798:Genes, Bacterial / Q000502:physiology*; D007709:Klebsiella / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003505:Cyclodextrins; D011134:Polysaccharides; C008315:maltodextrin