Ultraviolet (UV) causes skin disorders by inducing reactive oxygen species (ROS) and inflammation. Therefore, oral intake of ergothioneine (ERGO), which has antioxidant and anti-inflammatory activities, may protect from UV-induced skin damage since ERGO is efficiently absorbed from diet via its specific transporter OCTN1/SLC22A4 expressed in various tissues including epidermis . In the present study, we investigated the effect of novel strain of Pleurotus sp. (NPS), which highly contains ERGO, on UVB-induced skin damage in mice and human keratinocyte HaCaT cells. First, HR-1 hairless mice fed with 2.5% NPS containing diet or control diet were irradiated with UVB for 10 weeks. In the NPS ingested group, ERGO concentrations were ~130 µg/g tissue in the epidermis and ~40 µM in the plasma. In the skin, an oxidative marker 8-OHdG measured by ELISA and expression of TNF evaluated by western blotting were significantly lower than those in control group. In undifferentiated HaCaT cells, OCTN1 was detected in both plasma membranes and intracellular compartment. [3H]ERGO was taken up in HaCaT cells in a time-dependent manner, and transfection of siRNA targeted to OCTN1 remarkably suppressed the uptake. ERGO-containing NPS significantly suppressed intracellular ROS induced by UVB treatment. Taken together, NPS ingestion may lead to a high ERGO distribution to the skin, suppressing UVB-induced oxidative stress and inflammation possibly via protective effect exerted by ERGO taken up by its specific uptake transporter OCTN1 in keratinocytes.