L-Glutamic Acid Fermentation with Molasses

Abstract

the correlation of the fatty acid composition of each cell fraction and the extracellular accumulation of L-GA was carried out as follows: 1. Five factors-pH, temperature, the con-centration of lysozyme, the addition of in-organic ion and EDTA-were studied in order to determine the conditions suitable for the cell wall lysis of the L-GA-producing bacteria. The result was that the cell wall of M. ammoniaphilum-A was undoubtedly attacked to some extent, but was doubtful to be completely broken down by the lysozyme. On the contrary, the cell wall lysis of M. ammoniaphilum-C was observed with electron microscope and phase-contrast microscope to be completed under the following conditions: Bacterial cell 4.5mg/ml (as dry weight) m/20 Na-phos- Lysozyme phate buffer (pH 6.2) emp. 30°C 14 hr. Lysozyme 0.33mg/ml NaNO3 0.3Mm/ml, EDTA 0 2. Each of the C strain grown in the glucose-biotin (biotin level: 2.5γ/1 and 20 γ/1) media and beet molasses-POEFE (POEFE level: free and 0.15 g/dl) media was submitted to the cell wall lysis under the above men-tioned conditions. After removal of the cell wall fraction by 6, 000×g, the protoplast was ground by quartz. The cell membrane fraction and “the proto-plasm fraction” were fractionated at 12, 000×g, to prevent the contamination of the cell mem-brane fraction with the protoplasm. Each fraction was hydrolyzed and methylated before being submitted to the gas chromato-graphy. The correlation of the resulting fatty acid composition of each fraction and the extra-cellular accumulation of L-GA was discussed. From these results, following thesis was proposed: A large amount of extracellular accumulation of L-GA took place when the ratio of Sat. FA./Unsat. FA. in the cell mem-brane fraction was above 1 and the ratio in the cell wall and the protoplasm had no direct relation to the L-GA accumulation.