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I Urabe

Publications and source records attributed to I Urabe.

60 records · Page 4Linked to original sources
PubMed1978

Effect of hydrophobicity of acyl groups on the activity and stability of acylated thermolysin.

1. Normal carboxylic acids of different hydrophobicities and similar chain lengths were prepared and used for the modification of amino groups of thermolysin (EC 3.4.24.4). They were 4,7,10,13-tetraoxatetradecanoic acid, 4,7,10-trioxatetradecanoic acid, 4,7-dioxatetradecanoic acid and 4-oxatetradecanoic acid. 2. The modified enzymes were isolated by gel filtration. They had 6--7 acyl groups per molecule. Acylation of amino groups with 4-oxatetradecanoic acid and tetradecanoic acid made the enzyme insoluble. 3. The most hydrophilic enzyme derivative had similar enzyme activity and higher heat resistance than the native enzyme. The most hydrophobic derivative showed lower Km (50%) and V (40%) values for proteinase activity and lower heat resistance than the former derivative. The trioxa-derivative had intermediate characteristics. The results are discussed with respect to effects on stability and activity of the enzyme.

Acylation↗
PubMed1976

Role of calcium ions in the thermostability of thermolysin and Bacillus subtilis var. amylosacchariticus neutral protease.

The stabilizing effect of calcium ions on thermolysin and Bacillus subtilis var. amylosacchariticus neutral protease has been investigated. Calcium and zinc ions were removed from the proteases by gel filtration over Sephadex G-25 equilibrated with metal chelating agents. Using these enzymes with different metal content, heat inactivation kinetics were studied at various temperatures. Removal of calcium ions caused a sharp decrease in thermostability and diminished the values of the activation enthalpy (deltaH*) and entropy (deltaS*) for heat inactivation. There was little difference in stability between thermolysin containing 0.3 g-atom/mol and B. subtilis neutral protease containing 1.4 g-atoms/mol. Calcium binding isotherms of the proteases were obtained by equilibrium gel chromatography with various concentrations of free calcium ions. Thermolysin had four independent calcium binding sites with an identical intrinsic binding constant (K) of 2.0 X 10(4) M-1. B. subtilis neutral protease had four independent sites. The K value for three sites was 1.1 X 10(5) M-1 and the binding constant for the other site was 1.5 X 10(3) M-1. There was little difference in total free energy change for calcium binding between these proteases. From these results it is concluded that the stabilizing effect of calcium on these enzymes is almost equal, and the extra thermal stability of thermolysin is likely to come from its polypeptide chain structure.

Bacillus subtilis↗
PubMedDate not supplied

Bond strengths of single-bottle dentin adhesives to caries-affected dentin.

There is concern that some acidic conditioners may not be strong enough to adequately etch sclerotic or caries-affected dentin. The hypothesis that was tested was that there were no significant differences in the bond strengths of single-bottle bonding systems to normal or caries-affected dentin, regardless of the strength of the phosphoric-acid conditioner. Extracted teeth with coronal caries extending into mid-dentin were prepared by grinding the occlusal surface flat. This left a central region of caries-affected dentin surrounded by normal dentin. The One-Step bonding system was used to bond dentin following etching with 10 or 32% phosphoric acid. The Single Bond system was used after etching dentin with 10 or 35% phosphoric acid. After 24 hours in water, serial vertical sections were made through the bonded teeth to create slabs 0.7 mm thick. Each tooth yielded four to five slabs, some of which included normal dentin, while others included caries-affected dentin. Each slab was trimmed into an hourglass configuration to limit the test area to normal or caries-affected dentin. The results obtained with One-Step following etching with 10% phosphoric acid showed lower (P < 0.05) tensile bond strengths to caries-affected dentin compared to normal dentin (36.9 +/- 8.0 MPa vs 47.7 +/- 6.5 MPa, respectively). This difference disappeared when using 32% phosphoric acid (49.7 +/- 6.1 MPa vs 45.0 +/- 7.2 MPa, respectively). Bonds made to caries-affected dentin with Single Bond were always lower than bonds to normal dentin regardless of the strength of the phosphoric acid. Scanning electron microscopy of polished cross sections sequentially challenged with acid and NaOCl revealed loss of the middle of the hybrid layers created by either bonding system in caries-affected dentin etched with 10% phosphoric acid. It is clear that 32-35% phosphoric acid is required to adequately etch caries-affected dentin in order to produce high bond strengths and well-infiltrated demineralized dentin.

Acid Etching, Dental↗