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.     

Epiderstatin and Its Related Glutarimide Antibiotics Inhibit the Cell Growth Induced by Mitogen Stimulation

To investigate the immunosuppressive effects of glutarimide antibiotics including a new antibiotic named epiderstatin, we tested these antibiotics for inhibition of the blastogenesis of mouse spleen cells induced by mitogen stimulation (concanavalin A or lipopolysaccharide). The inhibitory activity was measured by colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. Among the glutarimide antibiotics tested, epiderstatin and acetoxycycloheximide especially strongly inhibited the blastogenesis of mouse spleen cells induced by concanavalin A and lipopolysaccharide, however, selectivity between T and B lymphocytes was not observed.     

Solid State 13C-NMR Analysis of Cell Wall Components of Fusarium oxysporum

An abdominal fat accumulation complicated by high blood triglycerides is regarded as a risk factor of metabolic syndrome. Feeding powdered nacre, mother of pearl, from Pinctada maxima, resulted in reduced body weight, visceral fat amount, and blood triglyceride level without influencing the food intake, body length, or amount of muscular tissue, suggesting that nacre powder specifically could decrease visceral fat.     

Identification of d-Erythro-Dihydroneopterin Triphosphate as a Product of Guanosine Triphosphate Cyclohydrolase from Serratia indica

Guanosine triphosphate cyclohydrolase (EC 3.5.4.16) was previously shown to exist in two forms (GTP cyclohydrolase D-I and D-II) in Serratia indica IFO 3759, and they were homogeneously isolated. The present study deals with the characterization of their reaction products. A fluorescent product formed from guanosine triphosphate by GTP cyclohydrolase D-II was identified as 7,8-dihydroneopterin triphosphate by its absorption spectra, phosphate analysis and gas chromatography-mass spectrometry of the dephosphorylated trimethylsilyl derivative. After oxidation and dephosphorylation, the d-erythro configuration of the side chain was made clear by the elution profile on ECTEOLA-cellulose chromatography, Rf values on thin-layer chromatography and by biological activity to Crithidia fasciculata ATCC 12857. The fluorescent products from GTP cyclohydrolase D-I and D-II were indistinguishable.     

Purification and Some Properties of Lignostilbene-a,/i-dioxygenase Responsible for the Ca—Cp Cleavage of a Diarylpropane Type Lignin Model Compound from Pseudomonas sp. TMY1009

A novel dioxygenase, lignostilbene-a,β-dioxygenase (LSD), which catalyzes cleavage of the interphenyl double bond of lignin-derived stilbenes, was isolated. Four isozymes of LSD were separated from cell-free extracts of Pseudomonas sp. TMY1009 by ion-exchange chromatography on a DEAE- Toyopearl column. The major isozyme, LSD-I, was purified to electrophoretic homogeneity and characterized.

LSD-I cleaved the interphenyl double bond of l,2-bis(4′-hydroxy-3′-methoxyphenyl)ethylene with the optimum pH at 8.5. The Km of LSD-I was 11 μm for the stilbene and 110/iM for oxygen. The molecular weight of LSD-I, which is composed of two identical subunits, was estimated to be 94,000. LSD-I contained 1 g atom of iron per 1 mol of enzyme protein.     

Rapid Gas Chromatographic Determination of Micro-quantities of N-Methyl Carbamates as their 2, 4-Dinitrophenyl Derivatives

A rapid and sensitive method is presented for the determination of micro-quantities (1 to 100 μg) of N-methyl carbamates as dinitrophenyl methylamine (DNP-MA) by electron capture gas liquid chromatography (GLC). The method described is characterized by rapidity and simplicity of procedure due to an improvement made in the present investigation, i.e., hydrolysis of an N-methyl carbamate and dinitrophenylation of the resulting methylamine with 2, 4-dinitro-1-fluorobenzene (DNFB) were accomplished simultaneously in a single reaction mixture. A novel approach was also made to eliminate unreacted DNFB by conversion to dinitrophenyl glycine (DNP-glycine).     

Studies on the Mode of Action of Polyoxins

(1) Chitin-UDP acetylglucosaminyltransferase (E.C. 2.4.1.16., chitin synthetase) in the cell-free system from phytopathogenic fungus Piricularia oryzae, and effects of various polyoxins and related compounds on the enzyme activity were studied. Polyoxins A~M, polyoxin A derivatives, polyoxin C derivatives, 5′-amino-5′-deoxyuridine, uridine and thymidine inhibited equally the incorporation of N-acetylglucosamine (GlcNAc) from UDP-N-acetylglucosamine (UDP-GlcNAc) into chitin.

(2) Competition between the above inhibitors and UDP-GlcNAc was observed by kinetic studies. The Km for UDP-GlcNAc was determined to be 3.3 × 10^?3 m and the Ki values for polyoxins A~M, except polyoxin C, were found to be in the range of 3.3 × 10^?5 m to 3.4 × 10^?6 m. For polyoxin C, 5′-amino-5′-deoxyuridine and uridine, the Ki values of 2.7 × 10^?3 m, 8.0 × 10^?3 m and 3.0 × 10^?3 m were given, respectively. The inhibitor constants for other related compounds were also calculated.

(3) The values of binding affinity, ?ΔG, for formation of substrate- or inhibitor-enzyme complexes were calculated from the Km or Ki values. In addition, partial binding affinities, ?Δg, for certain moieties or groups of polyoxins were estimated from the ?ΔG. For instance, the ?ΔG values for UDP-GlcNAc and polyoxin L were 5.7 kcal/mole and 9.2 kcal/mole, respectively. And the ?Δg values for the nucleoside moiety (part I), the carbamylpolyoxamic acid moiety (part II) and the carboxyl group at C5′ position of polyoxin L were 5.2, 3.5 and 0.7 kcal/mole, respectively.

(4) From the results obtained, the mechanism of action and relation between chemical structure and competitive inhibition of chitin synthetase were discussed.

     

Formation of Pungent Principles in Fruits of Sweet Pepper,Capsicum annuum L. var. grossum during Post-harvest Ripening under Continuous Light

Pungent principles (Capsaicinoid(s)) were found to be produced in fruits of sweet pepper, Capsicum annuum L. var. grossum, during post-harvest ripening under continuous light. The initial formation was observed after 4 days’ ripening. After 7 days’ ripening, the capsaicinoids content in placenta increased to 12.9 μg per fruit, which was 2.5-fold of that in pericarp. No pungent principles were detected in fruits during ripening in the dark and in seeds under continuous light. In placenta, the formation of dihydrocapsaicin and nordihydrocapsaicin which are the vanillylamides of saturated branched fatty acids was higher than that of capsaicin which is the vanillylamide of an unsaturated one. Remarkable formation and accumulation of carotenoid were also observed during post-harvest ripening under continuous light.