Purification and characterization of a hemoglobin-binding outer membrane protein of<i>Prevotella intermedia</i>

Guan, Su-Min; Nagata, Hideki; Maeda, Kengo; Kuboniwa, Masae; Minamino, Naoto; Shizukuishi, Satoshi

doi:10.1111/j.1574-6968.2004.tb09607.x

Cited by 9 publications

(3 citation statements)

References 28 publications

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“…However, the binding activity of the outer membrane was quite low (Figure 1). Guan et al reported that hemoglobin binding activity was confirmed in the outer membrane in P. intermedia , and a protein responsible for the bin ding to hemoglobin was isolated from the outer membrane, its molecular mass was 60 kDa [21], however HbBP of P. nigrescens was 46 kDa. These discrepancies may be due to the difference of bacterial species, even if both species are rather close each other.…”

Section: Discussionmentioning

confidence: 99%

Hemoglobin binding activity and hemoglobin-binding protein of prevotella nigrescens

et al. 2010

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Prevotella nigrescens, lacking siderophores was found to bind to the hemoproteins. The binding was observed also in the envelope which was prepared by sonication of the cell. The binding occurred in the pH-dependent manner; the binding was observed below neutral pHs of the incubation mixtures but only slightly observed in the neutral and alkaline pHs. Furthermore, hemoglobin bound to the envelope was dissociated at high pHs buffers. Maximum amounts of hemoglobin bound to 1 mg envelope was 51.2 μg. Kd for the reaction at pH 5.0 was 2.1 × 10-10M (210 pM). From the dot blot assay, hemoglobin could bind to a protein solubilized from the envelope by a detergent, referred to as hemoglobin-binding protein (HbBP), then it was purified by the sequential procedures of ion exchange chromatography, affinity chromatography and isoelectric focusing. Molecular weight and isoelectric point of the HbBP were 46 kDa and 6.1, respectively.

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Section: Discussionmentioning

confidence: 99%

Hemoglobin binding activity and hemoglobin-binding protein of prevotella nigrescens

et al. 2010

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“…OMPs were extracted after selective removal of inner membrane proteins as previously described [14]. An exponentially growing culture (1 litre, O.D.…”

Section: Methodsmentioning

confidence: 99%

The anti-adhesive mode of action of a purified mushroom (Lentinus edodes) extract with anticaries and antigingivitis properties in two oral bacterial pathogens

Signoretto

Marchi

Bertoncelli

et al. 2014

BMC Complement Altern Med

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BackgroundIn previous works we have shown that a low-molecular-mass (LMM) fraction from mushroom (Lentinus edodes) homogenate interferes with binding of Streptococcus mutans to hydroxyapatite and Prevotella intermedia to gingival cells. Additionally, inhibition of biofilm formation of both odonto- and periodonto-pathogenic bacteria and detachment from preformed biofilms have been described for this compound. Further purification of mushroom extract has been recently achieved and a sub-fraction (i.e. # 5) has been identified as containing the majority of the mentioned biological activities. The aim of this study was to characterise the bacterial receptors for the purified mushroom sub-fraction #5 in order to better elucidate the mode of action of this compound when interfering with bacterial adhesion to host surfaces or with bacteria-bacteria interactions in the biofilm state.MethodsCandidate bacterial molecules to act as target of this compound were bacterial surface molecules involved in cell adhesion and biofilm formation, and, thus, we have considered cell wall associated proteins (CWPs), teichoic acid (TA) and lipoteichoic acid (LTA) of S. mutans, and outer membrane proteins (OMPs) and lipopolysaccharide (LPS) of P. intermedia.ResultsFifteen S. mutans CWPs and TA were capable of binding sub-fraction #5, while LTA did not. As far as P. intermedia is concerned, we show that five OMPs interact with sub-fraction # 5. Capacity of binding to P. intermedia LPS was also studied but in this case negative results were obtained.ConclusionsBinding sub-fraction # 5 to surface molecules of S. mutans or P. intermedia may result in inactivation of their physiological functions. As a whole, these results indicate, at molecular level, the bacterial surface alterations affecting adhesion and biofim formation. For these antimicrobial properties, the compound may find use in daily oral hygiene.

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“…Protein profiles of bacteriophage were analyzed by onedimensional SDS-PAGE. Each purified bacteriophage (10 10 PFU/mL) sample was first solubilized by mixing an equal volume of 2× sample buffer solution (0.125 M Tris-HCl, 4% w/v SDS, 20% v/v glycerol, 0.01% w/v bromophenol blue, 10% v/v 2-mercaptoethanol) and boiled at 95 • C for 10 min (Guan et al, 2004). The solubilized sample was loaded into 5-15% Bullet PAGE One Precast Gel (Nacalai Tesque, Japan) and electrophoresed in 1× running buffer solution (0.25 mol/L Tris, 1.92 mol/L Glycine, and 10 g/L SDS) at 200 V for 30 min.…”

Section: Sds-page Analysismentioning

confidence: 99%

Isolation and Characterization of Six Vibrio parahaemolyticus Lytic Bacteriophages From Seafood Samples

et al. 2021

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Vibrio parahaemolyticus is a foodborne pathogen that is frequently isolated from a variety of seafood. To control this pathogenic Vibrio spp., the implementation of bacteriophages in aquaculture and food industries have shown a promising alternative to antibiotics. In this study, six bacteriophages isolated from the seafood samples demonstrated a narrow host range specificity that infecting only the V. parahaemolyticus strains. Morphological analysis revealed that bacteriophages Vp33, Vp22, Vp21, and Vp02 belong to the Podoviridae family, while bacteriophages Vp08 and Vp11 were categorized into the Siphoviridae family. All bacteriophages were composed of DNA genome and showed distinctive restriction fragment length polymorphism. The optimal MOI for bacteriophage propagation was determined to be 0.001 to 1. One-step growth curve revealed that the latent period ranged from 10 to 20 min, and the burst size of bacteriophage was approximately 17 to 51 PFU/cell. The influence of temperature and pH levels on the stability of bacteriophages showed that all bacteriophages were optimally stable over a wide range of temperatures and pH levels. In vitro lytic activity of all bacteriophages demonstrated to have a significant effect against V. parahaemolyticus. Besides, the application of a bacteriophage cocktail instead of a single bacteriophage suspension was observed to have a better efficiency to control the growth of V. parahaemolyticus. Results from this study provided a basic understanding of the physiological and biological properties of the isolated bacteriophages before it can be readily used as a biocontrol agent against the growth of V. parahaemolyticus.

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Purification and characterization of a hemoglobin-binding outer membrane protein ofPrevotella intermedia

Cited by 9 publications

References 28 publications

Hemoglobin binding activity and hemoglobin-binding protein of prevotella nigrescens

Hemoglobin binding activity and hemoglobin-binding protein of prevotella nigrescens

The anti-adhesive mode of action of a purified mushroom (Lentinus edodes) extract with anticaries and antigingivitis properties in two oral bacterial pathogens

Isolation and Characterization of Six Vibrio parahaemolyticus Lytic Bacteriophages From Seafood Samples

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