Inhibitory effects of autolysate of Leuconostoc mesenteroides isolated from kimoto on melanogenesis.

We investigated the inhibitory effects of the autolysate of Leuconostoc mesenteroides, a lactic acid bacterium isolated from sake mash, on melanogenesis in B16F0 murine melanoma cells and a human skin model. The autolysate: induced a decrease in melanin content in B16F0 murine melanoma cells and a 17%, 36%, 41% and 58% decrease in the human skin model by the application of 0.125, 1.25, 6.25, and 12.5 mg/tissue in total; decreased tyrosinase activity to 71%, 46% and 29% of control in B16F0 cells with 0.1, 0.2 and 0.4 mg/ml-medium respectively, but did not inhibit tyrosinase activity under cell-free conditions; decreased amount of tyrosinase in a dose-dependent manner from 74% with 0.1 mg/ml to 33% with 0.4 mg/ml; and decreased amount of tyrosinase mRNA to 80-90% with 0.2-0.4 mg/ml-medium. As the decrease in tyrosinase mRNA levels could not fully account for the reduction in protein, we suggest that the autolysate had post-transcriptional effects rather than transcription inhibition. Our results indicate that the autolysate of L. mesenteroides has potential therapeutic use as an effective anti-melanogenic agent.

Functional characterization of CCA1/LHY homolog genes, PpCCA1a and PpCCA1b, in the moss Physcomitrella patens.

The evolution of circadian clocks in land plants is not understood, because circadian rhythms have received little attention in plants other than angiosperms. We have characterized two genes, PpCCA1a and PpCCA1b, homologs of the Arabidopsis thaliana clock genes CCA1/LHY, from the moss Physcomitrella patens. PpCCA1a and PpCCA1b, together with angiosperm CCA1/LHY homologs, belong to the clock-associated single-myb gene family of green plants (including green algae and land plants). The accumulation of PpCCA1a and PpCCA1b mRNA showed rhythms with a period of approximately 1 day, phased as are those of angiosperm homologs, under 24 h light/dark cycles or in continuous dark. However, in marked contrast to angiosperm homologs, both genes showed arrhythmic profiles in continuous light. The timing of the PpCCA1b peak is determined by the time of the last light to dark transition, suggesting that the arrhythmicity in continuous light is due to dysfunction of the core clock. We generated single and double disruptants for PpCCA1a and PpCCA1b, and found that the double disruptants showed: (i) short periodicity and damped amplitude in the PpCCA1b rhythm, (ii) similar changes in the rhythmically expressed genes PpSIG5 and PpPRRa, and (iii) de-repression of PpCCA1b transcription levels, indicating negative feedback regulation. These observations indicate that the two genes are not merely structural homologs but also functional counterparts of CCA1/LHY. Together, our results illustrate similarities as well as divergence of the clock machineries between P. patens and A. thaliana, two distantly placed species in land plant phylogeny.