Melanin Inhibition Research Articles

Ganoderma proteins and their potential applications in cosmetics.

Ganoderma have been regarded as a traditional source of natural bioactive compounds for centuries and have recently been exploited for potential components in the cosmetics industry. Besides Ganoderma polysaccharides and triterpenes, multiple proteins have been found in Ganoderma. With the in-depth study of these proteins, various pharmacological functions of Ganoderma have become important in the discovery and development of new products. In the review, we summarized and discussed the kinds and characteristics of Ganoderma proteins, especially on fungal immunomodulatory proteins (FIPs) which can be potentially developed into cosmeceuticals or nutricosmetics and are a suitable target for production using established biotechnological methods. Furthermore, we discuss their pharmacological activities of the proteins with a focus on their pharmacological functions related to cosmetics, such as antioxidant activity, inhibition of melanin, antibacterial activity, and regulation of inflammatory mediators. Numerous other questions also are addressed before the proteins can be widely accepted and used as cosmetic additives.

Antioxidant and Antimelanogenesis Activities of Glyasperin A From Macaranga pruinosa Leaves

In our effort to find materials for drugs and cosmetics from tropical natural resources, we screened 21 methanol extracts from 7 Macaranga trees species ( Macaranga bancana, Macaranga gigantea, Macaranga hulle ttii, Macaranga pruinosa, Macaranga tanarius, Macaranga trichocarpa, and Macaranga triloba) for antioxidant and antimelanogenesis. The antioxidant and melanogenesis (tyrosinase enzyme assay and melanin inhibitor in B16 melanoma) assays were used to determine the activities. The fractionation and the isolation of active compound were done by various chomatographic methods and the structure was determined by spectroscopic analysis data. We isolated a prenylated flavonoid, named Glyasperin A, from M. pruinosa leaf. This compound showed a potency as antioxidant and inhibited melanin in B16 melanoma but not tyrosinase activity. These results showed that the methanol leaf extracts of M. pruinosa could be developed for cosmetic applications especially as a skin whitening agent.

Inhibitory effects of shell of Camellia oleifera Abel extract on mushroom tyrosinase and human skin melanin.

As an oil production byproducts, the shell of Camellia oleifera Abel (SC) is usually discarded in the dump. However, previous investigations suggested that the SC could provide valuable bioactive materials. The objectives of this study were to examine the ability of SC extract to inhibit in vitro tyrosinase activity and the melanin inhibition effects of cosmetic formulations containing SC 1,3-butanediol extract in human volunteers. The cell viability was determined using a WTT assay. A mushroom tyrosinase was used to evaluate the anti-tyrosinase activity of the SC extract. The placebo (no extract) or test (SC 1,3-butanediol extract) or positive control (kojic acid) cosmetic cream was applied on face of volunteers(30 female subjects) three times a day for 8weeks. The active compounds in SC extract were screened using liquid chromatography-high-resolution mass spectrometry (UHPLC-QTOF). The result showed that the cytotoxicity of SC extract is insignificant when the concentration of SC extract is below 160µg/mL. In addition, SC extract dose dependently inhibited tyrosinase activity and SC 1,3-butanediol extract possessed a stronger inhibitory activity than methanol extract and water extract. Clinical evaluations revealed that facial melanin levels of the volunteers receiving cosmetic formulations (containing SC 1,3-butanediol extract) were decreased 59% from baseline in 6th weeks, whereas the placebo group showed no effect. SC 1,3-butanediol extract was detected to contain 12 kaempferol compounds, significantly, kaempferol 3-O-[α-rhamnopyranosyl-(1→6)-β-glucopyranoside] and kaempferol-3,7-O-α-L-dirhamnoside are the major compounds. These results indicate that SC extract can be used as a natural skin-whitening agent in cosmetic products.

Inhibition of melanin production by sesquiterpene lactones from Saussurea lappa and their analogues

This research is aimed towards the discovery of a novel skin whitening agent from sesquiterpene lactones. We screened nine sesquiterpene lactones from Saussurea lappa root extracts, costunolide (1), α-, β-, γ-cyclocostunolides (2, 3, 4), santamarine (5), reynosin (6), arbuscirin A (7), dehydrocostus lactone (8), and 11β, 13-dehydrocostus lactone (9), for their melanin inhibitory activity in B16 melanoma cells. This study was focused on the inhibition of the extracellular melanin content. The screening results revealed that only α-cyclocostunolide (2) showed notable inhibition of the melanin production, particularly the extracellular melanin content, with an IC50 value of 5.75 μM. In addition, structure-activity relationship (SAR) of 12 sesquiterpene lactone analogues with reference to their melanin inhibitory activity was also investigated. However, none of the analogues were found to show any significant melanin inhibitory activity. To the best of our knowledge the above study is the first SAR investigation involving the melanin inhibitory activity of sesquiterpene lactones. It established that eudesmanolide-type lactone having a double bond between C3 and C4 showed the most efficient melanin inhibition. Additionally, the presence of a hydroxyl, ketone or ester moiety as the C1-substituent markedly decreased the melanin inhibitory activity. Thus, these findings suggest that α-cyclocostunolide (2) might be useful in the therapy of skin disorders.

795 Acetyl-arctigenin (Ac-ATG), a novel and safe skin lightening molecule

There is high demand to identify novel skin lightening actives that are effective and safe. Previously, it has been shown that arctigenin (ATG) found in A. lappa, inhibits melanogenesis. To explore this chemotype further and identify more potent skin lightening actives, several ATG derivatives were synthesized including acetyl-arctigenin (Ac-ATG). Utilizing murine melanoma cells, ATG and Ac-ATG both reduce melanin content, with Ac-ATG demonstrating increased potency. However, when measuring tyrosinase inhibition capacity, both ATG and Ac-ATG showed modest inhibitory activity (20-25%), while kojic acid (KA) reached almost 100% inhibition. Endothelin-1 (ET-1) is a key mediator for melanocytes in UVB induce pigmentation and melanosome transfer. Our results show both ATG and Ac-ATG inhibit ET-1 in human epidermal keratinocytes. We thus sought to evaluate the anti-melanogenesis activity of Ac-ATG and ATG when applied topically (MelanoDerm™). Results show that after 2 weeks, both 0.1% ATG and 0.1% Ac-ATG significantly reduce melanin production by 56% and 72% respectively. Moreover, 1% Ac-ATG results in an 84% inhibition of melanin, which is significantly greater than 2% KA (62% inhibition). Interestingly, when 0.1% Ac-ATG was combined with 2% KA, we observed an additive effect resulting in 96% reduction in melanin. Moreover, this effect is not observed when 0.1% ATG and 2% KA are combined (62% inhibition). Additional studies need to be performed to elucidate Ac-ATG’s mechanism of action. Lastly, safety assessment for Ac-ATG including, in vitro sensitization, skin and eye irritation, Ames test, phototoxicity and HRIPT have been performed and in all instances, Ac-ATG was found to be safe. Thus, Ac-ATG is a novel, safe depigmentation compound and our results suggest that it can be utilized as both a stand-alone skin lightning ingredient and/or in combination with other depigmentation compounds that function through different mechanisms.

Assignment of a dubious gene cluster to melanin biosynthesis in the tomato fungal pathogen Cladosporium fulvum.

Pigments and phytotoxins are crucial for the survival and spread of plant pathogenic fungi. The genome of the tomato biotrophic fungal pathogen Cladosporium fulvum contains a predicted gene cluster (CfPKS1, CfPRF1, CfRDT1 and CfTSF1) that is syntenic with the characterized elsinochrome toxin gene cluster in the citrus pathogen Elsinoë fawcettii. However, a previous phylogenetic analysis suggested that CfPks1 might instead be involved in pigment production. Here, we report the characterization of the CfPKS1 gene cluster to resolve this ambiguity. Activation of the regulator CfTSF1 specifically induced the expression of CfPKS1 and CfRDT1, but not of CfPRF1. These co-regulated genes that define the CfPKS1 gene cluster are orthologous to genes involved in 1,3-dihydroxynaphthalene (DHN) melanin biosynthesis in other fungi. Heterologous expression of CfPKS1 in Aspergillus oryzae yielded 1,3,6,8-tetrahydroxynaphthalene, a typical precursor of DHN melanin. Δcfpks1 deletion mutants showed similar altered pigmentation to wild type treated with DHN melanin inhibitors. These mutants remained virulent on tomato, showing this gene cluster is not involved in pathogenicity. Altogether, our results showed that the CfPKS1 gene cluster is involved in the production of DHN melanin and suggests that elsinochrome production in E. fawcettii likely involves another gene cluster.

Spiro-acridine inhibiting tyrosinase enzyme: Kinetic, protein-ligand interaction and molecular docking studies

Here, we evaluate spiroacridines as inhibitors of tyrosinase, a key enzyme to melanogenesis. For this purpose, the spiroacridines 3-(acridin-9-yl)-N-benzylidene-2-cyanoacrylohydrazide (AMTAC-01) and 3-(acridin-9-yl)-2-cyano-N-(4-metoxybenzylidene)-acrylohydrazide (AMTAC-02) were synthesized and their enzymatic inhibition types and mechanisms were investigated. In addition, the interaction of these compounds with the enzyme were studied by UV–Vis spectroscopy, spectrofluorimetry, 1H NMR titration as well as molecular docking. Spectroscopic results reveals that the acridine derivatives interact strongly (Ka ≅ 104 − 105 M−1) with the mushroom tyrosinase and the enzyme undergoes small structural modifications due to the interaction with AMTAC-01 compound. The interaction studies support the enzymatic inhibition results, which suggests that AMTAC-01 compounds inhibit the enzyme reversibly and follows a noncompetitive type (AMTAC-01) and mixed type (AMTAC-02) of inhibition. Nevertheless, AMTAC-02 (IC50 = 96.29 μM) inhibits the enzyme more effectively than AMTAC-01 (IC50 = 189.40 μM), which suggests a highly relevant role of AMTAC-02's methoxy group to the inhibition activity, which is confirmed by docking studies to mushroom tyrosinase. Docking also indicates this interaction to be absent in human tyrosinase. SignificanceBased on previous results which evidenced the relevant activity of two spiroacridinic compounds for cell growth inhibition against melanoma cells, here we improve our understanding about the spiroacridines in the biological media by exploring the molecular mechanism that govern the activities of these two compounds using mushroom tyrosinase (mTYR) enzyme as molecular target. The paper not only will have a major impact upon molecular mechanism that regulates melanin inhibition by spiroacridinic compounds, but also by guiding the search for enzyme inhibitors and the development of new anti-melanoma prophylaxis.

Soft corals collected from Jeju Island inhibits the \u03b1-MSH-induced melanogenesis in B16F10 cells through activation of ERK

In the present study, we first evaluated the melanin inhibitory effect of four crude 70% ethanol extracts separated from soft corals abundantly growing along the seawaters of Jeju Island, South Korea, including Dendronephthya castanea (DC), Dendronephthya gigantea (DG), Dendronephthya puetteri (DP), and Dendronephthya spinulosa (DS). Among the four ethanol extracts, the ethanol extract of DP (DPE) did not possess any cytotoxic effect on B16F10 cells. However, all other three extracts showed a cytotoxic effect. Also, DPE reduced the melanin content and the cellular tyrosinase activity without cytotoxicity, compared to the α-MSH-stimulated B16F10 cells. Specifically, DPE downregulated the expression levels of tyrosinase and microphthalmia-associated transcription factor by activating the ERK signaling cascade in α-MSH-stimulated B16F10 cells. Interestingly, the melanin inhibitory effect of DPE was abolished by the co-treatment of PD98059, an ERK inhibitor. According to these results, we suggest that DPE has whitening capacity with the melanin inhibitory effects by activating ERK signaling and could be used as a potential natural melanin inhibitor for cosmeceutical products.Graphical Molecular mechanisms associated with melanin inhibitory effect of soft coral extracts against α-MSH induced B16F10 cells

Melanin: Quantification and protection against oxidative stress in chromoblastomycosis agents.

Chromoblastomycosis (CBM) is a chronic cutaneous and subcutaneous infection caused by melanized fungal species. We quantified the extractable melanin of 77 strains of CBM agents distributed within five genera. Moreover, resistance to oxidative stress was evaluated in strains exposed or not to the melanin inhibitor tricyclazole. The median percentage of melanin mass extracted from dry fungal mass varied from 0.69 (Rhinocladiella similis) to 3.81 (Phialophora americana). Inhibition of melanin synthesis decreased survival rates to hydrogen peroxide. Together, these data highlight the importance of melanin in CBM agents.

Safety assessment, biological effects, and mechanisms of Myrica rubra fruit extract for anti-melanogenesis, anti-oxidation, and free radical scavenging abilities on melanoma cells.

Currently, the cosmetic and medical industries are paying considerable attention to solve or prevent skin damage or diseases, such as hyperpigmentation and oxidation and free radical damage. In this study, the effective compounds in Myrica rubra fruit were extracted and studied the biological effects of these M.rubra fruit extracts. In this study, we extracted M.rubra fruit using solutions with various ratios of water to ethanol (100:0, 50:50, 5:95) and studied the anti-melanogenesis, anti-oxidation and radical scavenging effects of these M.rubra fruit extracts on two melanoma cell lines: mouse melanoma (B16-F0) and human melanoma (A2058). The cytotoxicity, melanin synthesis, mushroom and cellular tyrosinase activities, enzyme kinetics, melanogenesis-related gene expression, melanogenesis-related protein secretion, radical DPPH scavenging activity and ROS inhibition after treatment with M.rubra fruit extracts were determined. The results showed that the water extract of M.rubra fruit was less cytotoxic to the melanoma cell lines, effectively inhibited melanin synthesis and tyrosinase activity and down-regulated the gene expression and protein secretion of MITF and TRP-1. In addition, the M.rubra fruit extracts also showed the abilities to scavenge DPPH free radicals and suppress ROS production. Finally, the effective compounds in the water extract were Myricetin-O-deoxyhexoside, Quercetin-O-deoxyhexoside, and Kaempferol-O-hexoside determined by LC/MS/MS assay. Overall, the water extract of M.rubra fruit is a safe and effective melanin inhibitor and anti-oxidant and can be applied widely in the fields of cosmetics and medicine.

Neural Stem Cells and Its Derivatives as a New Material for Melanin Inhibition

The pigment molecule, melanin, is produced from melanosomes of melanocytes through melanogenesis, which is a complex process involving a combination of chemical and enzymatically catalyzed reactions. The synthesis of melanin is primarily influenced by tyrosinase (TYR), which has attracted interest as a target molecule for the regulation of pigmentation or depigmentation in skin. Thus, direct inhibitors of TYR activity have been sought from various natural and synthetic materials. However, due to issues with these inhibitors, such as weak or permanent ability for depigmentation, allergy, irritant dermatitis and rapid oxidation, in vitro and in vivo, the development of new materials that inhibit melanin production is essential. A conditioned medium (CM) derived from stem cells contains many cell-secreted factors, such as cytokines, chemokines, growth factors and extracellular vesicles including exosomes. In addition, the secreted factors could negatively regulate melanin production through stimulation of a microenvironment of skin tissue in a paracrine manner, which allows the neural stem cell CM to be explored as a new material for skin depigmentation. In this review, we will summarize the current knowledge regulating depigmentation, and discuss the potential of neural stem cells and their derivatives, as a new material for skin depigmentation.

Anti-melanogenic effect of Prunus davidiana extract in melan-a melanocyte through regulation of OCA-2, TRP-1 and tyrosinase

Prunus spp. and locally available plants (used as folkloric medicine) were screened to find a novel and natural anti-melanogenic agent. Based on p-protein promoter reporter assay (PPRA) the candidate plants were screened in the quest for p-protein inhibitor. Expression profiling of key proteins revealed the molecular mechanism of the melanin inhibition as well as TEM analysis revealed melanosome structure. The screened plant extract through PPRA showed significant down regulation of p-protein, which led to melanin inhibition. Another key melanosomal protein like tyrosinase and TRP-1 was also found to be down-regulated. However, TRP-2 was not affected. TEM analysis of treated cells also revealed that the stage IV melanosomes were lowered in number compared to control. The present study shows the plants used in this study possess good anti-melanogenic properties. However, the P. davidiana has the highest anti-melanogenic property among screened plant extracts.

Acetazolamide Inhibits the Level of Tyrosinase and Melanin: An Enzyme Kinetic, In Vitro, In Vivo, and In Silico Studies.

Melanin is the major factor that determines skin color and protects from ultraviolet radiation. In present study we evaluated the anti-melanogenesis effect of acetazolamide (ACZ) using four different approaches: enzyme kinetic, invitro, invivo and in silico. ACZ demonstrated significant inhibitory activity (IC50 7.895±0.24μm) against tyrosinase as compared to the standard drug kojic acid (IC50 16.84±0.64μm) and kinetic analyses showed that ACZ is a non-competitive inhibitor without cytotoxic effect. In invitro experiments, A375 human melanoma cells were treated with 20 or 40μm of ACZ with or without 50μm of l-DOPA. Western blot results showed that ACZ significantly (P<0.05) decreased the expression level of tyrosinase at 40μm. Zebrafish embryos were treated with 10, 20 or 40μm of ACZ and of positive control kojic acid. At 72h of treatment with ACZ and kojic acid, ACZ significantly (P<0.001) decreased the embryos pigmentation to 40.8% of untreated embryos at the dose of 40μm of ACZ while kojic acid decreased only 25.0% of pigmentation at the same dose of kojic acid. In silico docking were performed against tyrosinase using PyRx tool. Docking studies suggested that His244, Asn260 and His85 are the major interacting residues in the binding site of the protein. In conclusion, our results suggest that ACZ is a good candidate for the inhibition of melanin and it could be used as a lead for developing the drugs for hyperpigmentary disorders and skin whitening.

Role of Melanin in Colletotrichum falcatum Pathogenesis Causing Sugarcane Red Rot

Melanin is one of the well-known secondary metabolite produced by fungi and other organisms which has a major role in pathogenicity. Our earlier studies established positive correlation on the production of secondary metabolites such as toxins, enzymes and melanin in relation to pathogen virulence and disease expression. In the present investigation, melanin induction as one of the factor in differentiating pathogen virulence was proved by extraction and quantification. The role of melanin in the pathogenesis of Colletotrichum falcatum has been studied in detail under the influence of melanin inhibitor, tricyclazole. Effect of tricyclazole on C. falcatum pathogenesis revealed that it is inhibitory to conidial germination, appressorium formation and its melanisation at lower concentrations. However, efficacy of tricyclazole was increased with increase in concentration in reducing C. falcatum growth, sporulation and symptom production in leaves and stalks. Finally, the presence of melanin in C. falcatum was confirmed by amplifying three melanin biosynthesis genes, viz. PKS1, SCD1 and THR1 at molecular level. The results confirm that melanin plays one of the important roles in C. falcatum pathogenesis and melanin inhibiting compounds have definite role in disease suppression by external application.

Liposomal delivery of ferrous chlorophyllin: A novel third generation photosensitizer for in vitro PDT of melanoma

BackgroundCutaneous melanoma (CM) has substantially increased among Caucasian populations in the past few decades. This increased the number of CM deaths throughout the world. Pigmentation of melanoma reduces the efficacy of photodynamic therapy (PDT). Third generation photosensitizers (PSs) are characterized by improved targeting to the diseased tissue and reduced systemic side effects. This study is directed towards synthesis and characterization of liposomes encapsulating sodium ferrous chlorophyllin (Fe-CHL) and assessing its efficacy as a PS in PDT of melanoma. MethodsPhenylthiourea (PTU) was used as a melanin synthesis inhibitor. PDT has been applied on de-pigmented melanoma cells using liposomes-encapsulated Fe-CHL. Cell death mechanisms after PDT were evaluated. ResultsTreatment of melanoma cells with 200μM of PTU for 48h provided 49.9% melanin inhibition without significant cytotoxicity. Transmission electron microscope (TEM) results proved an increase in the cellular uptake of liposomes by increasing incubation period from 6 to 24h via endocytosis with preferential accumulation in the mitochondria and the nucleus. Following de-pigmentation, PDT was applied resulting in LC50 of 18.20 and 1.77μM after 24 and 48h incubation with liposomes-encapsulated Fe-CHL respectively and exposure to 56.2J/cm2 monochromatic red laser of wavelength of 652nm. Mechanism of cell death of Fe-CHL mediated PDT was found to be a combination of both apoptosis and necrosis. ConclusionsLiposomes could be efficiently employed as a potential sustained release delivery system in the Fe-CHL-mediated PDT of de-pigmented melanoma.

Melanogenesis inhibitors from Coix lacryma-jobi seeds in B16-F10 melanoma cells

An adenine derivative, 9-β-D-glucopyranosyl adenine, reported for the first time from a natural source, in addition to nine known compounds were isolated from the seeds of Coix lacryma-jobi. Their structures were elucidated based on extensive spectroscopic and chemical studies. The isolated compounds and the ethanol extract have been assayed for melanin inhibition using B16-F10 melanoma cell line. The results of our study suggested the potential use of Coix lacryma-jobi seeds as a skin whitening agent and reveal the seeds to be a rich source of important phytochemicals with melanogenesis inhibitory activity. Among the isolated compounds, coixol (2) and 2-O-β-glucopyranosyl-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (8) exhibited potent melanogenesis inhibitory activity with no obvious melanocytotoxicity. The rest of the compounds showed weak to moderate activity.

Quantification analysis of protein and mycelium contents upon inhibition of melanin for Aspergillus niger: a study of matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS)

Mass spectrometry (MS) provides a simple discrimination method for microorganisms.

Rare ginsenoside Ia synthesized from F1 by cloning and overexpression of the UDP-glycosyltransferase gene from Bacillus subtilis: synthesis, characterization, and in vitro melanogenesis inhibition activity in BL6B16 cells

BackgroundGinsenoside F1 has been described to possess skin-whitening effects on humans. We aimed to synthesize a new ginsenoside derivative from F1 and investigate its cytotoxicity and melanogenesis inhibitory activity in B16BL6 cells using recombinant glycosyltransferase enzyme. Glycosylation has the advantage of synthesizing rare chemical compounds from common compounds with great ease. MethodsUDP-glycosyltransferase (BSGT1) gene from Bacillus subtilis was selected for cloning. The recombinant glycosyltransferase enzyme was purified, characterized, and utilized to enzymatically transform F1 into its derivative. The new product was characterized by NMR techniques and evaluated by MTT, melanin count, and tyrosinase inhibition assay. ResultsThe new derivative was identified as (20S)-3β,6α,12β,20-tetrahydroxydammar-24-ene-20-O-β-D-glucopyranosyl-3-O-β-D-glucopyranoside (ginsenoside Ia), which possesses an additional glucose linked into the C-3 position of substrate F1. Ia had been previously reported; however, no in vitro biological activity was further examined. This study focused on the mass production of arduous ginsenoside Ia from accessible F1 and its inhibitory effect of melanogenesis in B16BL6 cells. Ia showed greater inhibition of melanin and tyrosinase at 100 μmol/L than F1 and arbutin. These results suggested that Ia decreased cellular melanin synthesis in B16BL6 cells through downregulation of tyrosinase activity. ConclusionTo our knowledge, this is the first study to report on the mass production of rare ginsenoside Ia from F1 using recombinant UDP-glycosyltransferase isolated from B. subtillis and its superior melanogenesis inhibitory activity in B16BL6 cells as compared to its precursor. In brief, ginsenoside Ia can be applied for further study in cosmetics.

Laccase Gene Sh-lac Is Involved in the Growth and Melanin Biosynthesis of Scleromitrula shiraiana.

Scleromitrula shiraiana causes the popcorn disease in mulberry trees resulting in severe economic losses. Previous studies have shown that melanin may play a vital role in establishing the pathogenicity of fungi. In the present study, we identified the melanin produced in S. shiraiana belongs to DHN melanin by gas chromatography-mass spectrometry, and cloned the laccase Sh-lac, a potential DHN melanin biosynthesis gene from S. shiraiana. We obtained two stable Sh-lac silenced transformants using RNAi, ilac-4 and 8 to elucidate the DHN melanin biosynthetic pathway in S. shiraiana. The melanin production of ilac-4 and ilac-8 was significantly reduced, and their vegetative growth was also suppressed. Results such as these led to a proposal that Sh-lac played a key role in DHN melanin formation in S. shiraiana and may function differentially with other melanin biosynthetic genes. The inhibition of melanin was accompanied by the decrease of oxalic acid and the adhesion of hyphae was impaired. Our results indicated that laccase was an important enzyme in the synthesis of melanin and might play a critical role in the pathogenicity of S. shiraiana.

Effect of inhibition on tyrosinase and melanogenesis of Agastache rugosa Kuntze by lactic acid bacteria fermentation.

This work presents the first report that A. rugosa could have tyrosinase and melanogenesis inhibition and that its activities also be improved by fermentation with Lactobacillus rhamnosus and Lactobacillus paracasei. It was found that the tyrosinase and melanogenesis inhibition was correlated with antioxidant activity of acacetin, the major biologically active substances in A. rugosa. we pursued an improvement in tyrosinase and melanogenesis inhibition of A. rugosa extract by fermentation process. A. rugosa was extracted by lactic acid fermentation process; we measured antioxidant activities and tyrosinase and melanogenesis inhibition of A. rugosa extracts. In particular, reducing power of the extract from fermentation process (FE) was measured as 0.562 (O.D.), whereas reducing power of the extracts from 70% ethanol extraction (EE) was lower than the FE as 0.496 (O.D.). Polyphenols and flavonoids in the FE were higher than the EE: 69.3 mg/g vs. 60.5 mg/g, and 187 mg/g vs. 138 mg/g. The FE was estimated as 51.04% tyrosinase inhibition and 41.88% for the EE. Similarly, melanin inhibition in melanocyte B16F10 was observed as 66.60% vs. 42.23% for the FE and EE. The increase in tyrosinase and melanogenesis inhibition activity was confirmed by high elution of acacetin through fermentation process such as 289.97 mg/100 g vs. 198.04 mg/100 g in the FE and EE. These results indicate that tyrosinase and melanogenesis inhibition activities of the extracts should be associated with antioxidant activity because acacetin is known to have strong antioxidant activity, which can also positively affect whitening activities.