Mass Spectra of Postalotin and Some Related Compounds

The mass spectra of pestalotin and some related 4-methoxy-5,6-dihydro-α-pyrones, each containing a substituent at the C–6 position, have been studied. The prominent peak was observed commonly at m/e 127 and was proved characteristic of the pyrone ring.     

13C-NMR Studies on Halomethylvinyl Chrysanthemic Acids and Esters-Spin-lattice Relaxation Time and 13C-1H Coupling Constants

New types of stable chrysanthemic acids and esters were synthesized, and their ¹³C-NMR were examined and fully analyzed. The configurations of the cyclopropanecarboxylic acid and halomethylvinyl group were reflected on the spin-lattice relaxation time of the substituted methyl carbon involved in their structure. The long-range spin-spin coupling constants (³J_CH) correlated well to the NOE and T₁ measurements, which can generally be used to distinguish the geometry of the substituted double bond.     

Studies on the Compounds Produced by Molds

Three isocoumarin compounds (BV 1, 2 and 3) were isolated from the cultural broth of Aspergillus oniki 1784. BV 1 was mellein (3-methyl-8-hydroxy-3,4-dihydroisocoumarin). BV 2 and 3 were assigned to be 3-methyl-4,8-dihydroxy-3,4-dihydroisocoumarin, 3-methyl-3,8-dihydroxy-3,4-dihydroisocoumarin, respectively. These two compounds (BV 2, 3) were newly isolated. Also another component named BV 4 was proved to be 6-methylsalicylic acid.     

Assimilative Pathway of Methanol in Candida sp. Incorporation of 14C-Methanol, 14C-Formaldehyde, 14C-Formate and 14C-Bicarbonate into Cell Constituents

1. The incorporation of ¹⁴C-methanol, ¹⁴C-formaldehyde, ¹⁴C-formate and ¹⁴C-bicarbonate into a methanol-utilizing yeast, Candida N–16, was examined by paper-chromato-graphy and radioautography.

2. At the earliest time period examined, the highest percentage of radioactivity fixed from ¹⁴C-methanol or ¹⁴C-formaIdehyde into methanol-grown cells was found in fructose phosphate. The percentage distribution of radioactivity in fructose phosphate decreased as time elapsed. The radioactivity fixed from these compounds into glucose-grown cells was negligible compared with that fixed into methanol-grown cells.

3. The incorporation of ¹⁴C-formate into methanol-grown cells was extremely low. The highest percentage of radioactivity fixed for short time incubation was found in serine. The incorporation pattern of glucose-grown cells was similar to that of methanol-grown cells.

4. At the earliest time period, over 70% of radioactivity fixed from ¹⁴C-bicarbonate into methanol- or glucose-grown cells was found in aspartate.

5. These results suggest that in Candida N–16 methanol is specifically assimilated by a route with hexose phosphate as a primary stable intermediate.

     

Degradation of Cellular Phospholipids and Softening of Pressed Baker’s Yeast

Variations in lipid components of washings and homogenate of pressed baker’s yeast were investigated during the storage of pressed baker’s yeast at 30°C. Washings represents the substances which had leaked out from cells. Homogenate represents those contained in whole cells. Lipids in yeast washings increased toward softening, the phospholipids in yeast homogenate decreased continuously during storage. Two stages, an earlier period of storage (Stage I) and a later period of storage (Stage II) were observed in the degradation of phospholipids. Free fatty acid which was the main degradation product of phospholipid accumulated in Stage II, particularly at softening. The order in phospholipid degradation was PC>PE>PI + PS (PI>PS). Moreover, when washings of stored yeast at softening were assayed using ¹⁴C-acyl PC, the release of ¹⁴C-acyl fatty acid was observed.

These results suggest that phospholipids were degraded by some phospholipid-deacylating enzymes toward softening. From the results of lipid analysis, we inferred that the responsible enzymes were phospholipases.     

Synthesis of Theaspirone

β-Ionone (I) was oxidized to 2,3-epoxy-/β-ionone (II), which was converted to 2,3-dihydroxy-β-ionone (III) by acid treatment. III was reduced to 4-(1,2-dihydroxy-2,6,6-trimethylcyclohexan-1-yl)-2-butanol (V), which was converted, by oxidation, to cis- and trans-theaspirone (1-oxa-8-oxo-2,6,10,10-tetramethyl spiro-(4,5)-6-decene) (VII-A), (VII-B) and dihydroactinidiolide (2-hydroxy-2,6,6-trimethylcyclohexyliden-1-acetic acid lactone) (IX).     

Biochemical Studies on Rice Bran Lipase

Some chemical properties of the rice bran lipase were studied. The enzyme protein contained 14.98% nitrogen and consisted of 312 amino acid residues. It also contained a certain amount of lipid. The amino-terminal amino acids of the enzyme protein were shown to be glutamic acid and the carboxyl-terminal amino acids to be glycine and serine. The treatment of the enzyme protein with 8 m urea containing 1×10^?3m EDTA (ethyl-enediaminetetraacetic acid) seemed to cause dissociation of the subunits of the enzyme protein. From this observation and the results of the terminal amino acids analysis, it was presumed that the enzyme protein was composed of at least two types of subunits.     

Studies on Biochemistry of the Thiobacilli

In the oxidation of thiosulfate at pH 4.5 tetrathionate was formed as an intermediate, and the thiosulfate-oxidizing enzyme was active in acidic pH range in contrast to the enzyme of T. thioparus and Thiobacillus X.

Phosphate did not seem to affect the oxidation of thiosulfate but rather affect the conversion of tetrathionate. In the absence of phosphate, tetrathionate, which was produced from thiosulfate oxidation, seemed to accumulate without undergoing further conversion.

Quantitative oxidation of tetrathionate to sulfate was achieved with freshly harvested cells of T. thiooxidans; pH optimum for the oxidation of tetrathionate by the washed cells was 2~3, and the activity fell markedly at pH above 3.5.

Tetrathionate might be enzymatically dismuted to pentathionate and trithionate under anaerobic conditions with crude extracts of T. thiooxidans; pH optimum for the reaction was about 2.7 and the activity fell strikingly at pH 4.7. The formed trithionate might be further hydrolyzed to thiosulfate and sulfate.