Ganoderma lucidum, a species of the class Basidiomycetes, attracts international attention due to the wide variety of its biological activities and great potential as cosmetic ingredient, such as skin care cosmetics including ‘skin-whitening’. However, there is little information available regarding the tyrosinase inhibitory activity. Therefore, the objectives of this study were to investigate the chemical composition and tyrosinase inhibitory activity of the G. lucidum. To isolate active single compound from G. lucidum (GASC), we conducted ethanol extraction and chloroform fractionation. In addition, we assayed the inhibitoty effect of tyrosinase activity and melanin biosynthesis in B16F10 mouse melanoma cells. In the present study, we identified a GASC, which exhibited inhibitory effects of cellular tyrosinase activity, the protein expression of cellular tyrosinase and tyrosinase-related protein-1, 2. In additional, microphthalmia-associated transcription factor (MITF), as known as crucial regulator of melanogenesis-related genes, was down-regulated by treatment with GASC in a concentration-dependent manner. GASC exhibited significant inhibition of tyrosinase activity and melanin synthesis in B16F10. The finding that melanogenesis inhibitory effect of GASC will contribute to facilitate various approaches of this mushroom for use in skin whitening products.
The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges. fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylationsites(Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1(Asn-454), fvLac-2(Asn- 437andAsn-455), fvLac-3(Asn-111andAsn-237), and fvLac4 (Asn-402andAsn-457). In addition, the first 19–20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction(RT-PCR) analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes.
The 35.6-mb genome contained 12,218 predicted protein-encoding genes and 287 tRNA genes assembled into 11 scaffolds corresponding with the 11 chromosomes of strain KACC 42780. Based on presence of copper binding region and signal peptide, 8 laccase genes (Fv Lac-1, Fv Lac-2, Fv Lac-3, Fv Lac-4, Fv Lac-5, Fv Lac-6, Fv Lac-7, Fv Lac-8) were selected in F.velutipes genome. The laccase genes of F.velutipes ranged from 1,300 to 1,700 base pair in their size. In addition, the molecular mass and PI values of laccase genes ranged from 50 to 62 kDa and from 3 to 11, respectively.
In laccase activity assays, the highest laccase activity (3unit/ml)was shown in medium with 0.5mM of CuSO4 on 9th day. However, laccase activity was severely decreased in medium with 1mM and 2mM of CuSO4 was drastically decreased from day 3 through day 9. Interestingly, laccase activity in the medium without CuSO4 showed higher level than in the medium with 0.25, 1 and 2mM on both 6th and 9th day.
RT_PCR showed the highest transcription level with 0.25mM and 0.5mM of CuSO4 supplementation on day 3 and on day 9, respectively. In addition, the transcription level of Lac-1 increased depending on concentration of CuSO4 supplementation.
Ganoderma lucidum has a long history for traditional medicine in Asian countries. However, taxonomy of Ganoderma species remains controversial, since it was initially classified on the basis of morphological characteristics. Recently, Ganoderma sichuanense was proposed as a re-proposed name for Ganoderma lucidum in China. Likewise, all of the Korean G. lucidum were clustered into 1 group together with G. sichuanense from China, when a phylogenetic analysis was undertaken based on the ITS rDNA sequences of the Ganoderma species. Furthermore, G. lucidum from Europe and North America were clustered into different group. Based on these results, re-consideration of naming Korean cultivated G. lucidum is needed.
Flammulina velutipes is a popular edible basidiomycete mushroom found in East Asia and is commonly known as winter mushroom. Mushroom development showing dramatic morphological changes by different environmental factors is scientifically and commercially interesting. We have sequenced the 35.6-megabase pair genome of F. velutipes KACC42780 using a next generation sequencing (NGS) approach. Then, HiSeq2000 sequencing system was used for comparing gene expression of F. velutipes genome at three different developmental stages (mycelium, primordium, and fruit body). Ninety one percents (10,116 genes) of the 10,999 predicted genes are expressed in at least one developmental stage. In addition, analysis of expressed genes in three different developmental stages showed that only 0.25–1.5% (28–165) of the total expressed genes (10,013) were uniquely expressed in a single developmental stage, whereas 83.6% (9,198) was shared in all stages.
In the present study, the genetic diversity of 69 Ganoderma species from various regions was determined by different molecular markers, including the internal transcribed spacer (ITS) rRNA, a partial β-tubulin gene, and mitochondrial small-subunit ribosomal (SSU) rDNA gene as well as randomly amplified polymorphic DNA (RAPD) analysis. The size of the ITS rDNA regions and mitochondrial SSU rDNA gene from different Ganoderma species varied from 625 to 673 bp and 656 to 2,040bp, respectively, and those of the partial β-tubulin gene sequence were 419 bp. Phylogenetic analysis based on the ITS region, the partial β-tubulin gene, and the mitochondrial SSU rDNA gene reveal that Ganoderma species are largely divided into two groups. Interestingly, most of the Ganoderma lucidum strains could be classified into 1 group, while other Ganoderma species divided into several groups (4 or 5 groups) in phylogenetic tree. One fragment unique to G. lucidum was selected from the RAPD profile and then sequenced. One primer pair (designated as KGS-F and KGS-R) based on this specific fragment was designed to amplify a 559 bp DNA fragment within the sequenced region. A single band with the expected size of 559 bp was observed from G. lucidum, except for G. lucidum strains from China, Canada, and Taiwan. This specific marker for G. lucidum from RAPD analysis, also supported by the phylogenetic analysis of the ITS, partial β-tubulin gene, and mitochondrial SSU rDNA sequences, will be useful for the PCR-based identification of G. lucidum in research applications as well as in the market.
Ganoderma have been used in traditional medicine for thousands of years in Korea, China and Japan as well as in other regions of the world. Ganoderma species is known to grow on hardwoods, conifers, palms, old and declining trees, decay dead wood and stumps. Several species is cause serious diseases of plantation and other economically important tropical crops. It also has been reported to prevent and treat immunological diseases, including tumorigenesis, control of blood glucose levels, modulation of the immune system, hepatoprotection, bacteriostasis, and more. In the present study, we examined the effects of various Ganoderma species, including Ganoderma lucidum and Ganoderma neo-japonicum on nitric oxide inhibition activities, β-hexosaminidase secretions, and DPPH free radical scavenging capacities to compare the anti-inflammatory, anti-allergic, and antioxidant effects.
In the present study, a phylogenetic analysis was undertaken based on the internal transcribed spacer (ITS) rDNA and partial β-tubulin gene sequence of the Ganoderma species. The size of the ITS rDNA regions from different Ganoderma species varied from 625 to 673 bp, and those of the partial β-tubulin gene sequence were 419 bp. Based on the results, a phylogenetic tree was prepared which revealed that Korean Ganoderma lucidum strains belong in a single group along with a G. lucidum strain from Bangladesh.
Apple pomace is a by-product from the apple processing industry and has the potential to support the growth of microorganisms. In this study, the effect of apple pomace on the growth rate of Pleurotus ostreatus mycelium was investigated. The mycelial growth dramatically increased by 34.5, 20 and 26% in solid culture, liquid culture, and solid-state fermentation, respectively, by adding 2.5% apple pomace. However, the growth of P. ostreatus mycelia was slightly inhibited by adding 5 or 10% compared to 2.5% apple pomace. Our findings reveal that apple pomace utilization can become a model for the valuable addition of similar wastes, and for the development of a solid-state fermenter.