Identification and characterization of a protease from Streptococcus oralis C104

Abstract

Streptococcus oralis is among the earliest colonizers of the tooth surface during plaque formation. As such, its enzymatic activities may influence ecologic succession on the tooth surface. In the current study, we use zymograms and preparative polyacrylamide gel electrophoresis to identify and purify a protease from S. oralis (sanguis) C104. Proteases from S. oralis C104 were detected in cell pellets at 133, 146 and 176 kDa as clear proteolytic bands on gelatin-substrate zymograms. Preparation of the major (146 kDa) protease were obtained by continuous-elution electrophoresis. The protease was active over the pH range of 7 to 9 with optimum activity between pH 8 and 9. Protease activity was inhibited by several serine protease inhibitors including phenylmethylsulfonyl fluoride, di-isopropyl-phosphofluoridate and aprotinin. The protease showed highest hydrolytic activity against azoalbumin and Bz-Pro-Phe-Arg-NA. Immunofluorescence studies with a polyclonal antiserum to the 146-kDa protease suggest it is present on the cell surface of S. oralis C104. Zymograms of cell pellets from other S. oralis strains as well as S. sanguis and Streptococcus mitis suggest that functionally similar proteases are elaborated by many early colonizers of the tooth surface.

Protease production byStaphylococcus epidermidisand its effect onStaphylococcus aureusbiofilms

“…). Aprotinin is a commonly used serine protease inhibitor (Lo & Hughes, ; Park et al ., ). Iodoacetamide and EDTA are used as cysteine and metalloprotease inhibitors, respectively (Jankiewicz & Bielawski, ; Keller et al ., ; Yin et al ., ; Hassanein et al ., ).…”

“…4). Aprotinin is a commonly used serine protease inhibitor (Lo & Hughes, 1996;Park et al, 2012b). Iodoacetamide and Co-isolation with S. epidermidis, protease production of S. epidermidis (the average absorbance per CFU mL À1 is given) and the ability of S. aureus to form biofilms (evaluated by CV staining) are also given.…”

SufA – a novel subtilisin-like serine proteinase of Finegoldia magna

“…Microbial proteinases can also inactivate important host immunedefence molecules, such as immunoglobulins (von PawelRammingen & Björck, 2003), proteins of the complement system (Chen & Cleary, 1990) and antimicrobial peptides (Schmidtchen et al, 2002;Sieprawska-Lupa et al, 2004;Thwaite et al, 2006). Moreover, several species of oral streptococci produce extracellular proteinases capable of degrading albumin (Lo & Hughes, 1996), immunoglobulin A (Plaut et al, 1974) and salivary proteins (Choih et al, 1979). Many bacteria-derived proteinases cleave human kininogens, resulting in the release of kinins, potent proinflammatory peptides (Herwald et al, 1996;Imamura et al, 2005;Scott et al, 1993; for a review see Imamura et al, 2004).…”

Amylase-Binding Protein B of Streptococcus gordonii Is an Extracellular Dipeptidyl-Peptidase

“…S. gordonii is well known to colonize endothelial surfaces, especially damaged heart valves, in cases of infective endocarditis (15). Extracellular proteases produced by oral streptococci have been shown to degrade host proteins including albumin (24), salivary proteins (4), casein (28), and gelatin and collagen (14). It has been suggested that such proteases enable nutrient acquisition during times of stress or slow growth, as is seen within the bacterial vegetation causing endocarditis or within biofilms (17).…”