不完全菌Aureobasidium pullulansの菌体内ヒアルロン酸類似物質の動態

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Aureobasidium pullulans classified under Fungi Imperfecti markedly accumulates a hyaluronic acid-like substance (HALS) in the cell depending on culture medium and culture age. With the ultimate object of elucidating the correlation between the amount of the accumulated HALS and the physiological functions of the fungi, the authors have researched into the variation of HALS concentration in the cells with time, and the acid distribution. Further, they have investigated several properties of the HALS isolated from the fungi, and have obtained the following results : 1. Pure HALS was isolated by hydrolyzing cell homogenate with papain, removing protein from the homogenate with TCA, and by treating the resulting solution with DEAE-sephadex A-25. The yield of HALS was 10 to 12 mg per gram of cells. The value of protein to HALS was 0.005 or below. 2. Uronic acid and amino sugar were detected by thin layer chromatography of the degradation products of isolated HALS which was obtained by treating the HALS with hydrochloric acid or hyaluronidase. Infrared absorption spectrum of isolated HALS is alsmot the same as that of commercial hyaluronic acid extracted from bovine vitreous humor. The homogeneity of the isolated HALS was determined by analyzing the ultracentrifugal patterns of the acid. Sedimentation coefficient of the acid was determined at 2.64. 3. The Maximum intracellular accumulation of HALS in the fungi occurred after cultivation at 30℃ for 48 hours. After that, the accumulation of HALS gradually decreased up to 72 hours, and then increased again.4. The change in concentration of the intracellular HALS corresponded to the culture phase of the fungi in which the vegetative cells changed into chlamidospore-like cells.5. The accumulation of intracellular HALS was affected by the presence of yeast extract used as a nitrogen source in the medium, and by the concentration of glucose used as carbon source. That is to say, the intracellular HALS was accumulated in medium unfavoravble for the growth of the fungi.6. Intracellular HALS was detected in relatively large amounts in the cell wall fraction obtained from cell homogenate centrifuged at 7,400×g for 30 min. and was detected in almost equal amounts in both the supernatant and the precipitate on centrifugation at 140,000 ×g for 30 min. Though the weight of the precipitate was only 2.5% of the cell homogenate, the concentration of HALS contained in it was about 21.2% of the total intracellular HALS. The fraction containing a large amount of HALS was found to contain a lot of particles which could be stained purple with toluidine blue.
社団法人日本生物工学会の論文
1975-09-25

不完全菌Aureobasidium pullulansの菌体内ヒアルロン酸類似物質の動態

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