Daily topical applications of the concentrate of sake (CS) have been shown to reduce epidermal barrier disruption in murine skin caused by ultraviolet B (UVB) radiation, while one of the components of sake, ethyl alpha-D-glucoside (alpha-EG), also reduces barrier disruption. We confirmed the effect of oral ingestion of various doses of CS on epidermal barrier disruption caused by UVB irradiation in hairless mice. Then, to identify the effective components, we quantitatively analyzed alpha-EG, organic acids, and glycerol, the main components of CS, and examined the effect of various concentration of each on barrier disruption. alpha-EG and organic acids showed comparable results to CS itself, and transepidermal water loss levels in murine skin were significantly decreased as compared with the control. Furthermore, an investigation of the dose dependency of these agents was performed and the results showed the significant effectiveness of alpha-EG. In addition, red wine concentrate (WC) and beer concentrate (BC) were examined in order to confirm the unique effects of CS. Similar effects were not found with WC and BC.

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An autonomously replicating plasmid is expected to increase the frequency of Aspergillus transformation. To construct this type of plasmid, we developed a rapid method of re-isolating autonomously replicating plasmids from Aspergillus transformants. Transformants grown in MM medium under selective pressure for 1-2 days were converted to protoplasts with a cell wall lytic enzyme (e.g. Yatalase). The protoplasts were lysed with phenol/chloroform followed by precipitation with ethanol. The total DNA was treated with RNaseA, re-precipitated with PEG, and then used to transform E. coli. These re-isolated plasmids were mainly the plasmid monomer.