Sakura-cha (salted cherry blossom tea) is a Japanese tea that is traditionally served at celebrations such as wedding ceremonies. The production of Sakura-cha includes the immersion of cherry blossom flowers in Japanese plum vinegar, and through this process, the byproduct (plum vinegar extract of cherry blossom) is obtained. In this study, the antioxidant activity of the plum vinegar extract of cherry blossom was examined. The plum vinegar extract of cherry blossom had a greater superoxide anion scavenging activity compared with red wine, which is a well-known strong antioxidant. Liquid chromatography-mass spectrometry analysis showed that cyanidin-3-glucoside, cyanidin-3-rutinoside, and caffeic acid were the major components in the phenolic extract prepared from plum vinegar extract of cherry blossom, and they possessed superoxide anion scavenging activity. Caffeic acid is mainly responsible for the scavenging activity of phenolic extract; the contributions of cyanidin-3-glucoside and cyanidin-3-rutinoside were minor.

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and quadrupole formation (M2X+ + X-e M,X,, W,, or M+ + MX; + M,X,, KL, ; KL, = KL, 5) in addition to t h e ion pair formation (M+ + X-e MX, KL, in t h e concentration range (0.4-6.0) x lo-, mol dm-3. Surprisingly, a great enhancement in quadrupole formation for LiCF3C02 and LiC2F5C0, was observed in propylene carbonate with t h e highest relative permittivity ( E , = 64.4 at 25°C) of all t h e solvents. For trifluoroacetate, t h e limiting molar conductivity (Ao = 72.65) given by t h e Shedlovsky analysis C(O.4-4.0) x mol dm-3] was much larger than that CAO,ca,c = 26.373 calculated by Kohlrausch's additivity law with strong electrolytes. Lithium pentafluoropropionate gave a similar excess in t h e A. value. Computer simulations showed an increase in t h e Shedlovsky A. value with increase in t h e quadrupole formation constant. At t h e same time, t h e apparent association constant (M+ + X-+ MX, K,) calculated by Shedlovsky analysis was 10 times larger t h a n t h e ion-pair formation constant (Ka, in propylene carbonate (owing to strong quadrupole formation) and was much smaller than t h e Kg, value in the other solvents (mainly owing to strong triple-ion formation). A distinct triple-ion formation from tributylammonium trifluoroacetate or tributylammonium pentafluoropropionate was observed in benzonitrile. Causes of t h e failure in t h e Shedlovsky analysis have been discussed from t h e standpoint of higher-ion aggregates.Conductometry has proved a powerful method to examine the species present in solutions. Higher-ion aggregates, such as triple ions and quadrupoles from uni-univalent salts could form in solvents with lower relative permittivities. Fuoss and Kraus'*2 proposed triple-ion formation by electrostatic forces in very low permittivity media. Many conductometric investigations have been performed to examine solute-solvent and solute-solute interactions in non-aqueous solution^.^-^ The * -CMICX12 ' Standard deviation of the relative error.

Agarase genes of non-marine agarolytic bacterium Cellvibrio sp. were cloned into Escherichia coli and one of the genes obtained using HindIII was sequenced. From nucleotide and putative amino acid sequences (713 aa, molecular mass; 78,771 Da) of the gene, designated as agarase AgaA, the gene was found to have closest homology to the Saccharophagus degradans (formerly, Microbulbifer degradans) 2-40 aga86 gene, belonging to glycoside hydrolase family 86 (GH86). The putative protein appears to be a non-secreted protein because of the absence of a signal sequence. The recombinant protein was purified with anion exchange and gel filtration columns after ammonium sulfate precipitation and the molecular mass (79 kDa) determined by SDS-PAGE and subsequent enzymography agreed with the estimated value, suggesting that the enzyme is monomeric. The optimal pH and temperature for enzymatic hydrolysis of agarose were 6.5 and 42.5ºC, and the enzyme was stable under 40ºC. LC-MS and NMR analyses revealed production of a neoagarobiose and a neoagarotetraose with a small amount of a neoagarohexaose during hydrolysis of agarose, indicating that the enzyme is a β-agarase.