The high-molecular-mass amylase (HMMA) ofGeobacillus stearothermophilusATCC 12980 interacts with the cell wall components by virtue of three specific binding regions

    loading  Checking for direct PDF access through Ovid


The complete nucleotide sequence encoding the high-molecular-mass amylase (HMMA) ofGeobacillus stearothermophilusATCC 12980 was established by PCR techniques. Based on thehmmagene sequence, the full-length rHMMA, four N- or C-terminal rHMMA truncations as well as three C-terminal rHMMA fragments were cloned and heterologously expressed inEscherichia coli.Purified rHMMA forms were used either for affinity studies with the recombinant (r) S-layer protein SbsC (rSbsC), peptidoglycan-containing sacculi (PGS) and pure peptidoglycan (PG) devoid of the secondary cell wall polymer (SCWP), or for surface plasmon resonance (SPR) studies using rSbsC and isolated SCWP. In the C-terminal part of the HMMA, three specific binding regions, one for each cell wall component (rSbsC, SCWP and PG), could be identified. The functionality of the PG-binding domain could be confirmed by replacing the main part of the SCWP-binding domain of an S-layer protein by the PG-binding domain of the HMMA. The present work describes a completely new and highly economic strategy for cell adhesion of an exoenzyme.

Related Topics

    loading  Loading Related Articles