Studies on Oxygen Transfer in Submerged Fermentations

The unsteady-state gas analysis method for the determination of oxygen transfer rate in shaken cultures was applied to the glutamic acid fermentation. This method was essentially based on the measurement of oxygen tension of gas phase in flasks. Major intentions of this application were to clarify the problems of oxygen transfer, to establish the measure of aeration effectiveness and to design the best aeration condition in the glutamic acid fermentation. Correlations of the fermentation to the level of dissolved oxygen, the rate of oxygen transfer, overall oxygen transfer coefficient and oxygen demand of cells were examined. As a measure of aeration effectiveness, the rate of oxygen transfer actually occurring in the fermentation seemed to be preferable, and was found to be above 7 × 10^?7 mol/ml-min for attaining the maximum productivity. Extremely high or low oxygen supply resulted in less productivity.     

Responses of Isolated Plant Mitochondria to Photosynthesis Inhibiting Acylanilides

During experiments to elucidate the mode of action of photosynthesis inhibiting acylanilide type herbicides, the effects of various acylanilides on respiration and oxidative phosphorylation of isolated plant mitochondria were studied. The results showed that some acylanilides acted as uncouplers of oxidative phosphorylation: 1) Some stimulated the ADP-limited state 4 respiration of isolated mitochondria depriving them of their respiratory control ability during succinate oxidation. 2) Those which stimulated state 4 respiration interfered with oxidative phosphorylation to degenerate the P/O ratio.

The following relationships between chemical structure of acylanilides and their biological activities were demonstrated: 3) Among various ring-chlorinated propionanilides, the activity of 3′,4′-DCPA was especially prominent. 4) Almost all the side chain-substituted 3′,4′-dichloroacylanilides tested were effective. 5) Both chlorination of the 3 and 4 positions of the aniline moiety and acylanilide bonding were simultaneously required for an acylanilide to produce uncoupling activity. 6) DCMU was less effective than was 3′,4′-DCPA, both in stimulating state 4 and in degenerating the P/O ratio.     

A New Identification Method for Aliphatic Compounds Using Linear Equations of the GC Retention Index Value

The Retention Index (I) value and the different I values from two columns, carbowax 20M (20M) and OV101 (ΔI:I_20M ? I_OV101) of aliphatic compounds were calculated by several kinds of linear equations under programmed linear temperature conditions. The equations could be used to predict appropriate compounds as well as a database of I and ΔI values.     

Properties of Nucleoside Phosphorylase from Enterobacter aerogenes

The properties of uridine Phosphorylase (UPase) and purine nucleoside Phosphorylase (PNPase) at high temperature were investigated. Both enzymes were found to be distributed in a wide range of bacteria and were partially purified from Enterobacter aerogenes AJ 11125 by heat treatment, ammonium sulfate fractionation and column chromatographies onDEAE-cellulose and Sephadex G-150. The UPase was purified 109-fold, and it showed an optimum pH of 8.5 and optimum temperature of 65°C, and activity toward uridine, 2′-deoxyuridine, thymidine and uracil arabinoside but not cytidine. The Km values of UPase for uridine were 0.7 mm at 40°C and 1.8 mm at 60°C. The PNPase was purified 83-fold, and it showed an optimum pH of 6.8 and optimum temperature of 60°C, and significant activity toward purine arabinosides as well as purine ribosides. The Km values of PNPase for inosine were 0.8 mm at 40°C and 2.2 mm at 60°C.     

Studies on the Monoterpene Fraction of “Geranium Oil” from Pelargonium roseum Bourbon

Investigation has been carried out on the constituents of the monoterpene fraction of geranium oil from Pelargonium roseum Bourbon and, besides well-known components of the oil, some new components such as epoxylinalool, methyl heptenone, myrcene, limonene, p-cymene, citral and 2,2,6-trimethyl-6-vinyl-tetrahydropyran which was confirmed by deriving to cinenic acid, have been identified.     

Isolation of a Novel Auxin,Methyl 4-Chloroindoleacetate from Immature Seeds of Pisum sativum

Human casein was separated by gel filtration on a column of Sephadex G–200 with 0.1 m Tris buffer (pH 8.5) containing 1.0 m NaCl. The effluent which increased in turbidity at 25°C was centrifuged at 25,000 × g for 30 min and the precipitate was obtained as Fraction 6. After centrifugation, the effluent was separated into 5 elution fractions.

Disc gel electrophoretic patterns of each fraction showed occurrence of secondary bands other than major bands especially in Fractions 3, 4 and 5. The casein solutions unheated and heated at 100°C for 5 and 10 min were kept at 5°C for 5 days. No marked changes of electrophoretic pattern were observed among these casein solutions. However, when a casein solution heated at 100°C for 5 min was chroma to graphed under the same condition, secondary bands also appeared.     

Basic Aspects of Electrode Potential Change in Submerged Fermentation

The platinum electrode potentials relative to the standard half cell depended on a pH value, dissolved oxygen concentration, equilibrium constant and oxidation reduction potentials of the liquid The overall potential change in submerged fermentation gave no independent information on these individual factors A thermostatic and pH-static apparatus excluded influences of temperatures and pH values on the electrode pontentials If the determination was completed for short time duration, potentials were governed by the dissolved oxygen tension. While the oxygen concentration was maintained at a same level, redox potential changes became a dominant. This measurement of redox potential, which gave the concentration of extremely low dissolved oxygen that could not be detected by the membrane-coated oxygen electrode, was practically useful for the control of aerobic fermentation     

Synergistic Taste Effects of Some New Ribonucleotide Derivatives

Taste effects of six newly synthesized ribonucleotide derivatives, i.e., disodium salts of 2-methyl-5′-inosinic acid · 6H₂O, 2-ethyl-5′-inosinic acid · 1.5H₂O, 2-N-methyl-5′-guanylic acid · 5.5H₂O, 2-N-dimethyl-5′-guanylic acid · 2.5H₂O, 2-methylthio-5′-inosinic acid · 6H₂O and 2-ethylthio-5′-inosinic acid · 2H₂O, were studied. Stimulus thresholds (detection thresholds) of these derivatives ranged from about 0.02 to 0.006 g/100ml. Flavor-enhancing activities of them were 2.3 to 8.0 times larger than that of disodium 5′-inosinate · 7.5H₂O IMP) in the synergistic effect with monosodium glutamate. Furthermore, the quality of taste of all the derivatives was recognized to be the same kind to that of IMP.     

Conversions of Microbial Products in Relation to Oxygen Supply in Inosine Fermentation

On the basis of the ratio (cell’s respiration rate/maximum oxygen demand of cells, r_{a b}/K_rM) as a new criterion of oxygen supply, symptoms of oxygen deficiency was described in inosine fermentation. Conversions of the products were observed to occur in relation to the extent of oxygen deficiency. When oxygen demand of the cells was satisfied (r_{a b}/K_rM = 1.0), the cells accumulated exclusively inosine. Under limited oxygen supply at the value of r_{a b}/K_rM 0.5~0.9, on the other hand, inosine formation was inhibited and acetoin was the predominant product. When oxygen supply was limited more strictly at the value of r_{a b}/K_rM smaller than 0.3, the cells excreted 2,3-butyleneglycol as the main product.     

Ameloblastin in Hertwig’s Epithelial Root Sheath Regulates Tooth Root Formation and Development

Tooth root formation begins after the completion of crown morphogenesis. At the end edge of the tooth crown, inner and outer enamel epithelia form Hertwig’s epithelial root sheath (HERS). HERS extends along with dental follicular tissue for root formation. Ameloblastin (AMBN) is an enamel matrix protein secreted by ameloblasts and HERS derived cells. A number of enamel proteins are eliminated in root formation, except for AMBN. AMBN may be related to tooth root formation; however, its role in this process remains unclear. In this study, we found AMBN in the basal portion of HERS of lower first molar in mice, but not at the tip. We designed and synthesized small interfering RNA (siRNA) targeting AMBN based on the mouse sequence. When AMBN siRNA was injected into a prospective mandibular first molar of postnatal day 10 mice, the root became shorter 10 days later. Furthermore, HERS in these mice revealed a multilayered appearance and 5-bromo-2′-deoxyuridine (BrdU) positive cells increased in the outer layers. In vitro experiments, when cells were compared with and without transiently expressing AMBN mRNA, expression of growth suppressor genes such as p21^Cip1 and p27^Kip1 was enhanced without AMBN and BrdU incorporation increased. Thus, AMBN may regulate differentiation state of HERS derived cells. Moreover, our results suggest that the expression of AMBN in HERS functions as a trigger for normal root formation.