J. Cosmet. Sci., 65, 285–298 (September/October 2014) 285 Anti-skin-aging benefi ts of exopolymers from Aureobasidium pullulans SM2001 KYUNG HU KIM, SOO JIN PARK, JI EUN LEE, YOUNG JOON LEE, CHANG HYUN SONG, SEONG HUN CHOI, SAE KWANG KU, and SU JIN KANG Department of Histology and Anatomy, College of Korean Medicine, (K.H.K., S.J.P., J.E.L., C.H.S., S.H.C., S.K.K.), The Medical Research Center for Globalization of Herbal Medicine, (S.J.P., J.E.L., Y.J.L., C.H.S., S.H.C., S.K.K., S.J.K.), and Department of Preventive Medicine, College of Korean Medicine, Deagu Haany University, Gyeongsan, Republic of Korea (Y.J.L., S.J.K.). Accepted for publication July 6, 2014 Synopsis Background: There have been many attempts to search for affordable and effective functional cosmetic ingredients, especially from natural sources. Objectives: As research into developing a functional cos- metic ingredient, we investigated whether exopolymers from Aureobasidium pullulans SM2001 (E-AP- SM2001) exert antioxidant, antiwrinkle, whitening, and skin moisturizing effects. Methods: Antioxidant effects of E-AP-SM2001 were determined by measuring free radical scavenging capacity and superoxide dismutase (SOD)-like activity. Antiwrinkle effects were assessed through the inhibition of hyaluroni- dase, elastase, collagenase, and matrix metalloproteinase (MMP)-1. Whitening effects were measured by tyrosinase inhibition assay, and by melanin formation test in B16/F10 melanoma cells. Skin moisturiz- ing effects were detected by mouse skin water content test. Results: E-AP-SM2001 showed potent DPPH radical scavenging activity and SOD-like effects. Additionally, hyaluronidase, elastase, collage- nase, and MMP-1 activities were signifi cantly inhibited by E-AP-SM2001. We also observed that E-AP- SM2001 effectively reduced melanin production by B16/F10 melanoma cells and mushroom tyrosinase activities. Furthermore, signifi cant increases in skin water content were detected in E-AP-SM2001- treated mouse skin, as compared with vehicle-treated control skin. Notably, a mask pack containing E- AP-SM2001 showed a twofold more extensive moisturizing effect compared with one containing Saccharomycopsis ferment fi ltrate. Conclusions: Our results suggest that E-AP-SM2001 has adequate an- tiaging, antiwrinkle, and whitening benefi ts and skin moisturizing effect. These effects involve reducing hyaluronidase, elastase, collagenase, and MMP-1 activities, as well as inhibition of melanin production and tyrosinase activities. Therefore, the antioxidant E-AP-SM2001 may serve as a predictable functional ingredient. Address all correspondence to Sae Kwang Ku at gucci200@hanmail.net and Su Jin Kang at vegonia1@ hanmail.net. Kyung Hu Kim, Soo Jin Park, and Ji Eun Lee contributed equally to this work.

JOURNAL OF COSMETIC SCIENCE 286 INTRODUCTION Skin aging is characterized by clinical signs including wrinkles, irregular dryness, dyspigmen- tation, sallowness, deep furrows or severe atrophy, dehydration, telangiectases, premalignant lesions, laxity and a leathery appearance of skin (1). Skin aging is a complex biological process involving intrinsic factors (genetic factors, hormonal status, and metabolic reactions, such as oxidative stress) and extrinsic factors (chronic light exposure, pollution, ionizing radiation, chemicals, and toxins). A combination of these factors causes physiological alterations and progressive changes in each skin layer, and concomitant changes in skin appearance (2). Oxidative stress caused by reactive oxygen species (ROS) plays a pivotal role in the process of skin aging at the cellular level (3,4). ROS can block the formation of collagen, disrupt cellular renewal cycles, damage DNA, and stimulate the release of proinfl ammatory mediators (cyto- kines), which cause infl ammatory skin diseases (5–8). Additionally, ROS causes the depletion of antioxidant enzymes and destroy the cytoprotective defense mechanism by weakening an- tioxidant systems, thus rendering the skin susceptible to oxidative injury (9–11). To prevent and reduce skin aging, many people have used functional cosmetics that have a potent skin protective pharmacological effect (antiaging, whitening, antiwrinkle, mois- turizing, and skin protective effects) (12). Currently, there are various available ingredi- ents for functional cosmetics in the market. However, they have a number of limitations they are too expensive and have side effects, and their exact pharmacological mechanisms are not fully understood (13). Because of these factors, various investigations have con- tinuously attempted to search for affordable and effective functional ingredients, with fewer side effects, especially from natural sources (12,14). Purifi ed exopolymers from Aureobasidium pullulans SM2001 (E-AP-SM2001) comprise mostly β-1,3/1,6-glucan and other organic materials [amino acids, mono- or di-unsaturated fatty acids (linoleic and linolenic acids) and fi brous polysaccharides] (15). Recently, our team demonstrated that E-AP-SM2001.shows antiosteoporotic (16), anti-infl ammatory (17,18), and immunomodulatory effects (19). This fi nding prompted us to examine the protective effects of E-AP-SM2001 against skin aging in vitro and in vivo. The murine B16F10 cell line was used in this study, as it can produce melanin in response to α-melanocyte stimulating hormone (α-MSH) activation (20,21). The antioxidant effects of E-AP-SM2001 were determined by DPPH assay and by measuring superoxide dismutase (SOD)-like activity. Antiwrinkle effects were evaluated through the inhi- bition of hyaluronidase, elastase, collagenase, and matrix metalloproteinase (MMP)-1, because there is much evidence of close correlation between wrinkle formation and the loss of elasticity, collagenase, and MMP-1. Whitening effects were measured by tyrosinase inhibition assay and by measuring melanin formation in B16/F10 melanoma cells. To assess the skin moisturizing effects of E-AP-SM2001, skin water content was measured in Imprinting Control Region mice. METHODS AND MATERIALS CHEMICALS The solution and a viscous mask pack containing E-AP-SM2001 were supplied by Ari-Med Therapeutics (Daegu, Korea). Based on a previously reported analysis (15), the exopolymers of E-AP-SM2001 are known to consist of β-1,3/1,6-glucan (17%), β-1,4-glucan (18%),