ANTI-SKIN-AGING BENEFITS OF EXOPOLYMERS FROM AUREOBASIDIUM PULLULANS 293 the effects of oleanolic acid and E-AP-SM2001 on elastase activity were calculated as 29.36 ± 7.57 and 56.29 ± 23.06 μg/ml, respectively (Figure 2C and D). Collagenase activity was signifi cantly inhibited by treatment with oleanolic acid and E- AP-SM2001 at concentrations from 12.5 and 25 μg/ml, respectively. The IC50 values for the effects of oleanolic acid and E-AP-SM2001 on collagenase activity were calculated as 70.49 ± 21.08 and 107.21 ± 38.82 μg/ml, respectively (Figure 2E and F). MMP-1 activity was signifi cantly (p 0.01) inhibited by treatment with oleanolic acid and E-AP-SM2001 at concentrations from 12.5 and 25 μg/ml, respectively. The IC50 values for the effects of oleanolic acid and E-AP-SM2001 on collagenase activity were calculated as 71.93 ± 24.93 and 244.31 ± 74.71 μg/ml, respectively (Figure 2G and H). WHITENING EFFECTS OF E-AP-SM2001 Kojic acid and E-AP-SM2001 signifi cantly (p 0.01) inhibited mushroom tyrosinase activity at concentrations from 1.25 and 12.5 μg/ml, respectively. The IC50 values for the effects of kojic acid and E-AP-SM2001 on tyrosinase activity were calculated as 2.94 ± 1.04 and 53.17 ± 24.17 μg/ml, respectively (Figure 3A and B). Figure 3. Whitening effects of exopolymers from Aureobasidium pullulans SM2001 (E-AP-SM2001). Inhibi- tory activity of kojic acid and E-AP-SM2001 against tyrosinase (A and B) and melanin formation (C and D). Values are expressed as the mean ± SD of fi ve independent experiments. The vehicle control was set to 0%. IC50 indicates the concentration at which the percentage inhibition of tyrosinase activity and melanin forma- tion was 50%. a p 0.01 compared with control (MW test).

JOURNAL OF COSMETIC SCIENCE 294 Kojic acid and E-AP-SM2001 signifi cantly (p 0.01) inhibited B16/F10 melanoma cell melanin production at concentrations from 100 and 400 μg/ml, respectively. The IC50 values for the effects of kojic acid and E-AP-SM2001 on melanin production were calcu- lated as 610.39 ± 150.22 and 1,867.96 ± 288.80 μg/ml, respectively (Figure 3C and D). SKIN MOISTURIZING EFFECTS OF E-AP-SM2001 Compared with vehicle control skins, E-AP-SM2001 and SFF mask-treated mice skin showed an increase in water content from 30 min to 4 h after the end of application. The E-AP-SM2001-containing mask had a twofold more extensive moisturizing effect com- pared with the SFF-containing mask, and showed signifi cant (p 0.01 or p 0.05) in- creases in skin water content from 30 min to 8 h after the end of application (Table I). Compared with vehicle-treated control skins, the skin water contents of E-AP-SM2001- treated skins showed 26.82%, 32.79%, 24.76%, 14.28%, 6.62%, and 3.36% changes at 30 min and 1, 2, 4, 8, and 24 h after the end of application. Changes of 41%, 63%, 53.78%, 26.74%, 30.27%, 20.23%, and 4.21% were observed in E-AP-SM2001 mask- treated skins. Additionally, 25.69%, 28.50%, 21.93%, 13.12%, 3.63%, and 0.64% changes were detected in SFF mask-treated skins. DISCUSSION In the present study, we observed that E-AP-SM2001 appeared to be a suitable ingredient for functional cosmetics, exerting antiaging, antiwrinkle, whitening benefi ts and skin mois- turizing effect. E-AP-SM2001 being a natural antioxidant, extracted from Aureobasidium pullulans SM2001, constitutes an affordable ingredient. We suggest that E-AP-SM2001 is an attractive ingredient for anti-skin-aging benefi ts. E-AP-SM2001 showed potent DPPH radical scavenging effects and SOD-like activity. It is well documented that ROS causes oxidative damage of biological macromolecules, Table I Changes in Skin Water Content (%) Time after the end of application (hours) 0.5 1 2 4 8 24 Control 59.71 ± 3.23 52.26 ± 5.11 52.23 ± 3.40 52.88 ± 4.64 53.43 ± 4.31 53.13 ± 5.30 E-AP-SM2001 75.72 ± 5.00a 69.40 ± 7.60a 65.16 ± 5.00c 60.43 ± 6.63b 56.97 ± 4.04 54.92 ± 6.43 Mask 1 84.57 ± 4.94a 80.36 ± 7.22a 71.42 ± 8.75c 68.89 ± 7.76a 64.25 ± 6.34a 55.37 ± 6.34 Mask 2 75.05 ± 5.87a 67.15 ± 7.58a 63.68 ± 5.72c 59.82 ± 4.68b 55.37 ± 4.23 52.79 ± 3.70 Values are expressed as the mean ± SD. of eight mouse skins. Exopolymers from Aureobasidium pullulans SM2001 (E-AP-SM2001). Mask 1: mask pack containing E-AP-SM2001. Mask 2: mask pack containing Saccharomycopsis Ferment Filtrate (SFF). a p 0.01 compared with control (LSD test). b p 0.05 compared with control (LSD test). c p 0.01 compared with control (MW test).