Search PubMed⌕ Search

Biomedical subjects

I Karube

Publications and source records attributed to I Karube.

At least 127 records · Page 7Linked to original sources

Dye-coupled electrode system for the rapid determination of cell populations in polluted water.

We determined cell populations in polluted waters by using a fuel cell-type electrode. The electrode was constructed from a platinum anode, a silver peroxide cathode, and a membrane filter for retaining microorganisms. The principle of cell number determination is based on sensing a redox dye reduced by the microorganisms with the electrode. Sample solutions containing microorganisms were membrane filtered, and the resulting filter containing microbial cells was attached to the surface of a platinum anode. The electrode was immersed in phosphate buffer solution (0.05 M, pH 7) containing a redox dye (2,4-dichlorophenol-indophenol), and the current generated was measured. The response time of the electrode system was 10 to 20 min, and the current generated was proportional to cell populations above 10(4) cells/ml.

Bacillus subtilis↗

Electrochemical measurements of cell populations.

Determination of cell growth was carried out by a polarographic system. The system was constructed of two platinum electrodes, a saturated calomel electrode, and a thermistor electrode. Responses of the system to the dissolved oxygen, pH, and temperature were examined. Cell growth of S. cerevisiae and M. olivoasterospora was monitored continuously by this system. In addition, this polarographic system could be applied to the measurements of cell populations of the human cancer cell L-1210 and mouse leukocytes. The measureable range for these animal cells was approximately 10(3)--10(5) cells/ml.

Animals↗

Determination of microbial populations with piezoelectric membranes.

The ultrasonic determination of microbial population was carried out by using a new apparatus composed of two piezoelectric membranes. When the apparatus was immersed in a medium containing cells and an arbitrary voltage was charged on a piezoelectric membrane, an output voltage was obtained from the other membrane. The output voltage increased with increasing cell numbers. The linear relationships between the output voltage and microbial populations of Saccharomyces cerevisiae, Bacillus subtilis, and Klebsiella sp. were observed, and the output voltage was reproducible with a relative error of 6%. Furthermore, the cell population of S. cerevisiae in a fermentor could be continuously determined with this apparatus.

Journal Article↗

Microbial sensors for volatile compounds.

A microbial sensor consisting of immobilized microorganisms, a gas permeable membrane, and an oxygen electrode was prepared for the continuous determination of methyl alcohol. Immobilized methyl alcohol utilizing bacteria was employed for the sensor. The response time of the sensor was within 10 min by the steady-state method. A linear relationship was observed between the current decrease and the concentration of methyl alcohol below 22.5 mg l-1. The selectivity of the microbial sensor for methyl alcohol was satisfactory. Microbial sensors using Trichosporon brassicae for ethyl alcohol and acetic acid are also described. A microbial sensor consisting of immobilized nitrifying bacteria (isolated from activated sludges), a gas-permeable Teflon membrane and an oxygen electrode was prepared for the amperometric determination of ammonia. When the sensor was inserted in a solution containing ammonia, the current decreased to a steady-state with a response time of 4 min. The relationship between the current decrease and the ammonia concentration was linear up to 42 mg l-1. The minimum concentration for the determination was 3.5 mg l-1. The current decrease was reproducible within 4 per cent of relative error. The current output of the sensor was almost constant for over 10 days and 200 assays.

Acetates↗

Effect of alcohols on the collagen-phosphatidylcholine interaction.

The interaction of phosphatidylcholine dispersions with acid soluble collagen separated from the skin of one month-old swine was studied to define the conditions facilitating the association of the collagen with lipids. When acid soluble collagen and phosphatidylcholine dispersions were incubated in 75 mM citrate buffer of pH 3.7 at 25 degrees C, the reisolated collagen fibrils did not contain appreciable amounts of phosphatidylcholine. However, the presence of n-propanol greatly promoted the retention of phosphatidylcholine, the amount of phosphatidylcholine associated being nearly 30% of collagen on a weight basis under optimal conditions. In contrast, methanol, ethanol, isopropanol, and n-butanol did not appreciably enhance the association of phosphatidylcholine with collagen. A limited inhibition of phosphatidylcholine retention was observed upon addition of sodium chloride to the propanol medium. The interaction of phosphatidylcholine with acid soluble collagen decreased sharply when temperature was increased above 30 degrees C; almost no phosphatidylcholine-collagen association occured at 40 degrees C. It appears that the enhanced association in the presence of n-propanol is due to a looseing of the collagen triple helix that exposes hydrophobic sites necessary for the interaction. However, the conversion of the triple helical structure to the random coil conformation by heating prevents the association of phosphatidylcholine with acid soluble collagen.

Alcohols↗

Penicillin G production by immobilized whole cells of Penicillium chrysogenum.

Penicillium chrysogenum was immobilized in polyacrylamide gel prepared from 5% acrylamide monomers (85% acrylamide and 15% N,N'-methylene bisacrylamide). Penicillin produced from glucose by the immobilized mycelium was 17% of that produced by washed mycelium. However, the activity of penicillin production of the washed mycelium decreased with repeated use. On the other hand, the activity of the immobilized mycelium increased initially and decreased gradually with repeated use. The rate of oxygen uptake of the immobilized mycelium was about 30% of that of the washed mycelium. The immobilized mycelium required oxygen for the production of penicillin.

2-Aminoadipic Acid↗

Bacitracin production by whole cells immobilized in polyacrylamide gel.

Whole cells of Bacillus sp., bacitracin-producing bacteria, were immobilized in polyacrylamide gel prepared by using 5% total acrylamide (95% acrylamide monomer and 5% N, N'-methylenebis acrylamide). Bacitracin production by the immobilized whole cells was examined by using various fermentation media. In starch-bouillon medium, the initial activity of immobilized whole cells for bacitracin production was 20 to 25% that of an equivalent amount of washed cells. With successive utilizations in 1% peptone as reaction medium, activity of the immobilized whole cells increased gradually and reached a steady-state maximum having a value of 80 to 90% of the activity obtained initially with washed cells, whereas the washed cells lost most activity when utilized successively. What appeared to be growth of the bacteria in the gel was observed by electron microscopy. Therefore, the activation of immobilized whole cells during successive utilizations apparently resulted from the growth of whole cells in the gel, especially at the gel surface.

Acrylamides↗

Electrode system for the determination of microbial populations.

Determinations of microbial populations were carried out by using a new electrode system composed of two electrodes. Each electrode was constructed from a platinum anode and a silver peroxide cathode. The anode of the reference electrode was covered with cellulose dialysis membrane. The response time of the electrode system was 15 min in culture broth, and current differences between the two electrodes were proportional to populations of microbial cells in cultures of Saccharomyces cerevisiae and Lactobacillus fermentum. Current differences were reproducible; the average relative error was 5%. Furthermore, cell populations of S. cerevisiae in a fermentor could be continuously estimated by using this electrochemical method.

Journal Article↗

NAD recycling in the collagen membrane.

NAD recycling in the collagen membrane was investigated as follows: (1) Alcohol dehydrogenase and lactate dehydrogenase were co-immobilized in the collagen membrane and the rate of lactate production by immobilized enzymes was compared with that of free enzymes by using free NAD. An increased rate was observed in the case of immobilized enzyme. (2) The soluble high molecular weight derivatives of NAD (dextran-NAD) were immobilized in the collagen membrane with the two dehydrogenases and recycling of dextran-NAD in the membrane was examined. Lactate was produced by the membrane without adding free NAD. The interaction between the high molecular weight NAD derivatives and enzymes are also discussed.

Alcohol Oxidoreductases↗

Bacteriolysis by immobilized enzymes.

Bacteriolytic enzymes produced by Achromobacter lunatus were immobilized in collagen membrane. Intact bacteria such as Pseudomonas solanacearum, Xanthomonas oryzae, Staphylococcus aureus, and Pseudomonas aeruginosa were lyzed with the bacteriolytic enzyme-collagen membrane. Relative activity of the bacteriolytic enzyme-collagen membrane against Pseu. solanacearum was about 2% of that of native bacteriolytic enzymes. No difference in the optimum pH was observed between immobilized enzymes and native enzymes. The bacteriolytic enzymes in the collagen membrane were stable against sodium chloride which was an inhibitor of the native bacteriolytic enzymes. Xanthomonas oryzae and Pseu. aeruginosa were continuously lyzed by a reactor containing the rolled bacteriolytic enzyme-collagen membrane.

Alcaligenes↗

Aminoacylase pellets.

Aminoacylase was immobilized on the mycelium pellets of Aspergillus ochraceus by using albumin and glutaraldehyde. No difference in the optimum pH was observed between native aminoacylase and aminoacylase pellets. The aminoacylase pellets were stable in pH 4-8 but they were unstable in alkaline conditions. The aminoacylase pellets were more stable against heavy metal ions and inhibitors than native aminoacylase. However, the degree of the activation of aminoacylase with cobalt ion decreased with the immobilization. It was suggested that most of aminoacylase was covalently coupled to the mycelium with glutaraldehyde.

Amidohydrolases↗

Electric field control of lipase membrane activity.

Lipase (EC 3.1.1.3., from Pseudomonas sp.) was entrapped in collagen membrane containing liquid crystal (4-methoxybenzilidene-4'-n-butylaniline). The activity of the lipase-liquid crystal membrane at an applied voltage of 4 V was 3.4 compared to a membrane tested without imposition of an external electric field. A linear relationship was observed between the activity of the lipase-liquid crystal membrane and the current. The apparent Michaelis constant (K'm) of the lipase-liquid crystal membrane under electric field was identical to that of the membrane under ordinary condition. Activation of the lipase-liquid crystal membrane was observed repeatedly, i.e., activation in the presence of an electric field and reversion to a basal level upon removal of the field occurred cyclically. Activity control of immobilized enzymes is desirable for switching devices of a bioreactor. Possible mechanisms of the lipase activation by electric field are discussed.

Collagen↗

Photocontrol of urease activity in spiropyran collagen membrane.

1. Collagen fibrils were modified with beta-1-[3,3-dimethyl-6'-nitrospiro-(indoline-2,2'-2H-benzopyran)] propionic anhydride. 2. Urease (urea amidohydrolase, EC 3.5.1.5) was immobilized in spiropyran collagen membrane. The activity of the urease-spiropyran collagen membrane was found to increase in the dark and then decrease with visible light irradiation. 3. The optimum pH of the urease-spiropyran collagen membrane under visible light was lowered in the dark. 4. The apparent Michaelis constant (K'm) of the urease-spiropyran collagen membrane in the dark was almost the same as that under visible light. The apparent maximum velocity was increased in the dark. 5. The diffusion coefficient of urea through the spiropyran collagen membrane in the dark was 1.4 times that under visible light. However, the increase of the diffusion rate was not responsible for the activity increase of the urease-spiropyran collagen membrane.

Collagen↗