The kinetics and mechanism of shear inactivation of lipase from Candida cylindracea

Shearing experiments were conducted in a stirred tank reactor with 0.1% lipase solutions of Candida cylindracea. Inactivation of the lipase solutions were observed at various shear rates from 50 to 150 s(-1) after continuous shearing for ca. 30-240 min under optimal pH and temperature conditions. However, there was no shear stress denaturation of the lipase when it was subjected to shear stresses of 0.72-109.2 kg/m/s(2) and shear rate of 100 s(-1). In the presence of polypropylene glycol, the rate of denaturation of the lipase decreased by 93%. When the lipase solution was filled to the brim, the rate of denaturation of the lipase decreased by 97% compared to that when reactor was half-filled. The rate of denaturation of the lipase decreased by 61% when probes in the fermentor were removed. There was no significant difference in the rate of denaturation of the lipase under ambient conditions compared with that in the absence of oxygen, or in the absence of free metal ions. Recovery of lipase activity from the first hour of shearing was observed at a shear rate of 150 s(-1). The native lipase and the lipase which had recovered its activity showed similar pH profiles, temperature profiles, and activation energies. Temperature was found to have no effect in the rate of shear-induced denaturation of the lipase in the range 20 to 30 degrees C during shearing at 100 s (-1)and optimal pH. Above 30 degrees C, the rate of denaturation of the lipase increased drastically as a function of temperature. The significance of the findings in the de sign of reactor systems for hydrolysis or esterification of oils by lipase will be discussed.     

Diffusivity of oxygen into carriers entrapping whole cells

The effective diffusivity of oxygen, D(e), in Ca-alginate and PVA-SbQ gels was measured using a two-chamber vessel with a membrane between the two chambers. The effect of cell density, C(c), on D(e) in Ca-alginate gels was studied. The effective diffusivity of oxygen decreased with increasing cell density, to C(c) = 170 kg dry cells/m(3) gel. The dependency of D(e) on cell density was discussed in terms of a random-pore model. The model correlated well with experimental data, i.e., kD(e)/D(0) = 0.86(1 - 1.47 x 10(-3) C(c))(2). Here, k is the partition coefficient, and D(0) is diffusivity in water.     

Kinetic model of hydrocortisone 1-en-dehydrogenation by Arthrobacter globiformis

A kinetic model of the hydrocortisone-to-prednisolone transformation by Arthrobacter globiformis is constructed using the experimental data obtained in studies of this process. Besides adequately describing experimental data, the model allows one to determine the relation between hydrocortisone oxidation and the level of endogenous substrates in bacterial cells, and the relation between the saturating concentration of hydrocortisone in the enzymic system of bacteria and the content of endogenous substrates in their cells, as well as the regulation of the transmembrane potential and the activation by the uncouplers.     

Origin of changes in electrical impedance during the growth and fermentation process of yeast in batch culture

The electrical impedance of the culture medium shows complex changes during the growth and fermentation process of yeast, and this prevents its possible application for the monitoring of certain yeast activities. Clarification of the mechanism of such changes is thus essential for practical use. As a first step toward this aim, the impedance, yeast concentration, and pH of a batch culture medium were measured using special cells with two compartments and also the usual type of cell with one compartment. In the special cells, the yeast was cultured in one compartment only. Conducting ions and nonconducting substances diffused through an intermediate porous membrane sandwiched between the two compartments. The impedances of the two compartments were measured simultaneously by the four-electrode method. The main mechanism responsible for increasing the impedance was the conducting ions produced by the yeast extract added as a nutrient to the culture broth by certain nonconducting substances during the process of growth. The increase in the yeast concentration was also a minor factor increasing the impedance. These increases surpassed the impedance decrease caused by the increase of H(+) ions produced by some accumulated acidic substances, and the impedance thus increased.     

Dynamics of plasmid maintenance in a CSTR upon square-wave perturbations in the dilution rate

The effects of forced oscillations in the dilution rate on a population of Escherichia coli K12 harboring the plasmid pBR322 in a chemostat with a nonselective medium were studied. In the constant dilution rate control experiments, the percentage of plasmid-containing cells decreased after a long lag time. Eventually, the culture approached a population consisting of 100% plasmid-free ells. However, under forced perturbations of the dilution rate, the culture maintained a mixed population of plasmid-free and plasmid-carrying cells for a longer period of ime. An unstructured model was developed to describe the above observations. Our results indicate that transient conditions created by dilution-rate perturbations provide a favorable environment for the plasmid-carrying population. In addition, experiments with different cycling frequencies suggest that adaptation by the culture to these transient conditions will reduce or totally eliminate such an advantage.     

Isolation and preliminary characterization of a 2-chlorobenzoate degrading Pseudomonas

Pseudomonas sp. strain B-300, which is able to utilize 2-chlorobenzoic acid, was isolated from a soil sample by enrichment culture. This strain was shown to grow on 2-chlorobenzoic acid and to completely degrade the substrate with concomitant chlorine ion release. Concentrations of 2-chlorobenzoic acid higher than 0.5% (w/v) were toxic to the cells. Our study also suggested that in the presence of glucose, 2-chlorobenzoic acid is converted to catechol or chlorocatechol; these are in turn transformed to muconic and chloromuconic acid, respectively, suggesting a repression by glucose of some of the degradation pathway enzymes. A similar scheme was already described for 3-chlorobenzoate degradation by pAC25 plasmid.     

Change of incidence of antinuclear antibodies in serum of NOD mouse with aging

Extractable nuclear antigens (ENA) were prepared from liver of C57BL/6J mouse and analyzed by SDS PAGE Western-immunoblotting techniques. Some protein components of the ENA, with molecular weights of 94 K, 65 K, 32 K, and 26 K, reacted with antinuclear antibodies in the sera of NOD mice. Incidence of antinuclear antibodies in the sera of NOD mice with aging were measured by ELISA method using the ENA as antigen. The antinuclear antibodies were not detected in young NOD mice (10 weeks old). However, the incidence increased with aging and reached 100% in the female NOD mice of 40 weeks. In the male NOD mice, the incidence of antinuclear antibodies was delayed and low in comparison with that in female.     

Detection of nuclear protein antigens to antinuclear antibodies in serum of NOD mouse

Nuclear protein antigens to the antinuclear antibodies in serum of non-obese diabetic (NOD) mice were investigated. In the serum of diabetic NOD female mice (20 weeks old), the antinuclear antibodies were detected by indirect immunofluorescence assay using frozen sections of liver of C 57 BL/6 J or NOD mice as antigen. Nuclei were separated from the liver of C 57 BL/6 J mice and solubilized. Solubilized nuclear antigens were analyzed by SDS PAGE-Western immunoblotting techniques. Nuclear protein antigens with molecular weights of 26,000, 32,000 and 65,000 showed strongly positive reactions with the antinuclear antibodies in the serum of the NOD mouse.     

A computer program that draws pedigree charts for inbred strains of animals

We produced a computer program that draws pedigree charts for inbred strains of animals such as mice or rats. This program is composed of four subprograms, which are (1) inputting the data, (2) drawing pedigree charts, (3) listing the data which have been input, and (4) backup of the system and the data. Pedigree charts and lists of data can be displayed on a TV screen and printed out on the papers. Using this program, we drew the pedigree charts of the inbred strains of rats which we are maintaining by brother-sister inbreeding in our institute and found that there were three sublines in one of the strains, WKAH/Hkm, because of unsuitable maintenance. This program is very convenient to draw the pedigree charts and useful for checking the maintenance of inbred strains or the strains of animal models of human diseases.     

Measurement of lactose concentrations in milk of Microtus montebelli with methylamine reaction

Concentrations of lactose in milk of Microtus montebelli were measured with the method using with the methylamine reaction. In this method, a small amount of sample (0.5 g) was sufficient for assay, and reproducibility and sensitivity were excellent. This method was very useful for measuring of lactose concentrations in experimental small animals. The average lactose concentration in milk of Microtus montebelli was 1.57g/100g, considerably low in comparison with the value of ICR mice (2.70g/100g). The low concentration of lactose in milk was considered to be one of metabolic characteristics of Microtus montebelli as a herbivorous animal.