title: The high-molecular-mass amylase (HMMA) of Geobacillus stearothermophilus ATCC 12980 interacts with the cell wall components by virtue of three specific binding regions.
abstract: The complete nucleotide sequence encoding the high-molecular-mass amylase (HMMA) of Geobacillus stearothermophilus ATCC 12980 was established by PCR techniques. Based on the hmma gene 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 in Escherichia 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.
authors.affiliation: Department of NanoBiotechnology, University of Natural Resources and Applied Life Sciences, A-1180 Vienna, Austria.
authors.forename: Judith
authors.initials: J
authors.lastname: Ferner-Ortner-Bleckmann
authors.forename: Carina
authors.initials: C
authors.lastname: Huber-Gries
authors.forename: Tea
authors.initials: T
authors.lastname: Pavkov
authors.forename: Walter
authors.initials: W
authors.lastname: Keller
authors.forename: Christoph
authors.initials: C
authors.lastname: Mader
authors.forename: Nicola
authors.initials: N
authors.lastname: Ilk
authors.forename: Uwe B
authors.initials: UB
authors.lastname: Sleytr
authors.forename: Eva M
authors.initials: EM
authors.lastname: Egelseer
mesh_terms: D000681:Amylases / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism*; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics; D010457:Peptidoglycan / Q000378:metabolism; D011485:Protein Binding; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
chemical_list: D001426:Bacterial Proteins; D010457:Peptidoglycan; D000681:Amylases