Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Purification and Characterization of Extracellular Ice-nucleating Matter from Erwinia uredovora KUIN-3
Hidehisa KawaharaYoshinori ManoHitoshi Obata
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1993 年 57 巻 9 号 p. 1429-1432

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The extracellular ice-nucleating matter (EIM) produced by the ice-nucleating bacterium, Erwinia uredovora KUIN-3, was purified to apparent homogeneity by ultrafiltration, sucrose density-gradient ultracentrifugation, and gel filtration. The purified EIM was sphercial matter (0.2-0.4 μm) by examination using a transmission electron microscope. The ice-nucleating activity from the EIM was equal to that of the cells in this strain. It had become apparent that the EIM had class A (∼ -5°C), B (-5 ∼ -8°C), and C (-8°C ∼) structures as judged by its freezing difference spectrum in D2O versus H2O. The enzyme treatments by protease K, α-mannosidase, β-galactosidase, and phospholipase C decreased the activity of the EIM as well as of the cells. Further, the activities of class A and B from the EIM were decreased by treatment with diamine oxidase. Also, the activity of the purified. EIM was stable from pH 5.0 to 10.0 and at temperatures below 25°C. It was demonstrated that the components of the EIM were lipid (10%), protein (43%), saccharide (35%), and polyamine (12%).
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