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Fujisawa, Japan

Kato S.,Nagoya University | Ishihara T.,Nagoya University | Hemmi H.,Nagoya University | Kobayashi H.,Mercian Corporation | Yoshimura T.,Nagoya University
Journal of Bioscience and Bioengineering | Year: 2011

Alterations in d-amino acid concentrations and microbial community structures during the fermentation of red and white wines were analyzed to clarify the relationship between the occurrence of d-amino acids and the actions of fermentative microorganisms. Relatives of Saccharomyces cerevisiae and Oenococcus oeni were detected in red wine samples, and relatives of S. cerevisiae, O. oeni, and Gluconacetobacter saccharivorans were detected in white wine samples. The S. cerevisiae relatives were detected throughout the fermentation process, whereas the O. oeni relatives were detected at the late stage of fermentation in both the red and white wine samples. The G. saccharivorans relative was detected in the early stage of fermentation. The amino acid analysis showed that d-alanine, d-glutamic acid, and d-lysine were present in both the red and white wine samples. The concentrations of these d-amino acids increased as the fermentation continued, especially from the malolactic fermentation stage to the end of the fermentation processes. These increases seem to be linked to the presence of O. oeni relatives. © 2010 The Society for Biotechnology, Japan.

Shimazaki M.,Yamanashi University | Fujita K.,Yamanashi University | Kobayashi H.,Yamanashi University | Kobayashi H.,Mercian Corporation | Suzuki S.,Yamanashi University
PLoS ONE | Year: 2011

We report here that pink grape berries were obtained by a short insertion in the intron of the MybA1 gene, a gene that regulates grape berry color. Genetic variation was detected among the MybA1 genes from grapes cultivated worldwide. PCR analysis of the MybA1 gene demonstrated that the size of the MybA1 gene in the red allele differs among grapes. Oriental V. vinifera bearing pink berries has the longest MybA1 gene among grapes, whereas the shortest MybA1 gene was detected in occidental V. vinifera grapes. The nucleotide sequences of the MybA1 genes demonstrated that oriental V. vinifera has two additional gene fragments (44 bp and 111 bp) in the promoter region of the MybA1 gene in the red allele and another 33 bp fragment in the second intron of the MybA1 gene in the red allele. The short insertion in the intron decreased the transcription activity in the model system and retained MybA1 transcripts with unspliced intron in the total RNA. From the experiments using deletion mutants of the 33 bp short insertion, 16 bp of the 3′ end in the insertion is a key structure for a defect in splicing of MybA1 transcripts. Thus, a weakly colored grape berry might be a result of the short insertion in the intron of a color regulatory gene. This is new evidence concerning the molecular mechanism of the fate of grape berry color. These findings are expected to contribute to the further understanding of the color variation in grape berries, which is correlated with the evolutional events occurring in the MybA1 gene of grapes. © 2011 Shimazaki et al.

Mercian Corporation | Date: 2013-08-27

Wine; alcoholic fruit beverages.

Kasai N.,Toyama Prefectural University | Ikushiro S.-I.,Toyama Prefectural University | Shinkyo R.,Toyama Prefectural University | Yasuda K.,Toyama Prefectural University | And 5 more authors.
Applied Microbiology and Biotechnology | Year: 2010

The white-rot fungus Phanerochaete chrysosporium possesses biodegradative capabilities of polychlorinated dibenzo-p-dioxins (PCDDs). One hundred twenty yeast clones expressing individual P450s of P. chrysosporum (PcCYPs), generated in our previous efforts, were screened for transformation of dioxin, and 40 positive clones were obtained. Of these clones, six clones showed metabolism of 2-chloro-dibenzo-p-dioxin, and a microsomal PcCYP designated as PcCYP11a3 showed much higher activity than any other PcCYPs. The turnover numbers of hydroxylation activities of PcCYP11a3 toward 1-MCDD (58 min-1) and 2-MCDD (13 min-1) are more than 200 times higher than those of previously reported PcCYP65a2. In addition, PcCYP11a3 catalyzes hydroxylation of 2,3-dichloro-dibenzo-p-dioxin. To our best knowledge, PcCYP11a3 has the highest activity toward PCDDs among the known CYPs derived from microorganisms. Although PcCYP11a3 showed no detectable activity toward 2,7-dichloro-dibenzo-p- dioxin and 2,3,7-trichloro-dibenzo-p-dioxin, PcCYP11a3 is promising as a template whose activity would be enhanced by site-directed mutagenesis. © 2010 Springer-Verlag.

Izumikawa M.,Biomedicinal Information Research Center | Hashimoto J.,Biomedicinal Information Research Center | Hirokawa T.,Japan National Institute of Advanced Industrial Science and Technology | Sugimoto S.,Mercian Corporation | And 3 more authors.
Journal of Natural Products | Year: 2010

Proteasome assembling chaperone (PAC) 3 acts as a homodimer and plays an important role in proteasome formation. We screened JBIR-22 (1) as an inhibitor for protein-protein interaction (PPI) of PAC3 homodimer from our natural product library using a protein fragment complementation assay (PCA) with monomeric Kusabira-Green fluorescent protein (mKG) in vitro and found that 1 exhibited potent inhibitory activity against PAC3 homodimerization. Compound 1 showed long-term cytotoxicity against the human cervical carcinoma cell line, HeLa. This is the first report of a PPI inhibitor for proteasome assembly factors. © 2010 The American Chemical Society and American Society of Pharmacognosy.

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