Porphyromonas gingivalis培養上清中のアルギニン特異的プロテアーゼのアイソザイムの比較

By anion-exchange chromatography of the concentrated culture fluids of P. gingivalis W 50, two proteolytic enzymes, referred to as RGP-I and RGP-II were, eluted from column at the different NaCl concentrations in the eluant buffer. The two enzymes were purified until the preparations displayed single bands with sodium dodecyl sulfate-polyacryl amide gel electrophoresis (SDS-PAGE) by gel filtration and isoelectric focusing. Comparative studies between the two enzymes revealed that the same molecular masses (44 kDa) of RGP-I and II were obtained both with gel filtration and SDS-PAGE. However, the isoelectric points of RGP-I and II were 5.0 and 5.7, respectively. Both enzymes were thiol proteinases and were inhibited by sulfhydryl group blocking reagents, tosyl-L-lysine chloromethylketone, EDTA, leupeptin, L-trans-epoxy-succinylleucylamido-(4-guanidino) butane, and antipain. Both enzymes were activated by glycylglycine. RGP-I and RGP-II hydrolyzed synthetic substrates containing arginine in the P-1 positions, but those containing lysine in the same position were notcleaved

Additional Properties of a Bacteriocin-like Agent (Acnecin) of Propionibacterium acnes

Purified acnecin showed that it exerted bacteriostatic activity against susceptible indicator cells without adsorption on the cells, and its activity lost by treatment with Na-periodate or lysozyme. Acnecin inhibited also production of an extracellular enzyme (Ribonuclease) of acnecin-susceptible strain

Treponema denticolaの産生するコラゲナーゼ基質分解酵素の精製と性状

A peptidase hydrolyzing synthetic substrates of collagenase was purified from cell extract of an oral spirochete, Treponema denticola by sequential procedures including anionic ionexchange chromatography, gel filtration, and hydrophobic interaction chromatography. The purified enzyme was most active at pH 8.0 and was inhibited by p-chloromercuribenzoate or EDTA. This enzyme was active on Pz-peptide and Z-Gly-L-Pro-L-Leu-Gly-L-Pro. It was observed that Gly-L-Pro was released from the latter substrate, but proteins, including collagens, were not found to be hydrolyzed by this enzyme

capnocytophaga ochraceaのα-グルコシダーゼ;その局在性,可溶化,精製および性状

α-Glucosidase activity was detected in the crude extract of cell and envelope of Capnocytophaga ochracea ATCC 33596. The enzyme associated with the envelope was effectively dissolved by cetyltrimethylammonium bromide. The enzyme was purified from both fractions by combination of ammonium sulfate precipitation, ion-exchage chromatography, gel filtration, and isoelectric focusing. Properties of both enzymes were quite similar: they were basic proteins, metalloenzymes, and most active at pH7.0. They hydrolyzed only p-nitrophenyl-α-D-gluconopyranoside

エラスターゼ産生性Prevotella intermediaの菌体外ゼラチナーゼの分離と性状

Gelatinase in the culture supernatant of Prevotella intermedia was purified to homogeneity by the combined procedure of ethanol precipitation and three steps of chromatography. The enzyme was a cystein protease with a molecular mass of 45 kDa. The activity was inhibited by divalent metal chelators and its inhibition was recovered by the addition of Ca^. The optimum pH for activity was 7.0-7.5 and the enzyme was inactivated by heating at 60℃ for 10 min. It hydrolyzed actively azocoll and hide powder besides gelatin. Hydrolysis of type IV collagen, if not strong, was also observed

Isolation and Properties of Low Molecular Weight Antimicrobial Agents (Matrucin) from an Oral Bacterium Bacterionema matruchotii

Antimicrobial agents, designated Matrucin-1 and Matrucin-2, were isolated from culture supernatants of Bacterionema matruchotii IBN6, obained from dental plaque. They seem to be peptides with molecular weights less than 1,000. Matrucin-1 and Matrucin-2 were hardly soluble in polar solvents. Both inhibited the growth of various oral bacteria

Rothia dentocariosaとStreptococcus oralisとの共凝集反応

In the course of studies on assessment of coaggregation among oral indigenous bacteria, Rothia dentocariosa was found to coaggregate Streptococcus oralis and Streptococcus mitis. One heat-labile and one -stable binding sites on the bacterial surfaces may be responsible for coaggregation. The binding sites of S. oralis are destroyed by protease treatment. No inhibition of coaggregation activities was demonstrated in the presence of serum, saliva, sugars, and amino acids, however, EDTA inhibited the activities completely

Purification and Properties of Hyaluronidase (EC 4. 2. 2. 1) from an Oral Strain of Propionibacterium acnes

From a culture supernatant of P. acnes isolated from a lesion of periodontal disease, hyaluronidase was purified to homogeneity by the sequential procedures including ammonium sulfate precipitation, carboxy methy-cellulose column chromatography, and Sephadex G-100 gel filtration. Specific activity increased 1,027 fold and the recovery of the enzymatic activity was 12.4%. Molecular weight was determined to be 67,000 and isoelectric point was 7.2. Optimal pH for the activity was found at 5.5. The enzyme was inactivated by heating at 60℃ for 10 min. The purified hyaluronidase degraded hyaluronic acid, chondroitin, chondroitin sulfate A and chondroitin sulfate C. From the degradation products of these substrates, unsaturated disaccharides were detected by paper chromatography. When the rate of reaction of this enzyme against hyaluronic acid is supposed to be 100%, the corresponding values against chondroitin, chondroitin sulfate A, and C were 47%, 8%, and 8%, respectively. No degradation by this enzyme of heparin and heparan sulfate was demonstrated