Bioprospecting Endophytic Fungi for Production of Acid Stable Laccases and Tyrosinase Inhibitors for Their Development into User Friendly Skin Bleaching Formulations

Abstract

Skin lightening agents (SLA’s) are the agents which are incorporated into creams and lotions for development of cosmeceutical products for over the counter and therapeutical use. The market of SLA’s expected to reach US$ 31.2 billion by 2024. SLA’s inhibits the synthesis of melanin by acting upon the tyrosinase or they bleach the melanin which is already present in the cells. However, due to some side effects of the existing SLA’s, there is utmost need to explore alternative natural source. Endophytic fungi are endosymbionts which colonize the healthy living tissue of host plant. They are considered as reservoir of bioactive compounds which exhibits anti-cancer, anti-microbial and anti-fungal activities. The present study was based upon the screening and identification of tyrosinase inhibitor and laccase producing endophytic fungi. Initially, culture filtrates of 91 cultures isolated from Aegle marmelos , Cinnamomum sp. , Vitis vinifera were screened for Tyrosinase inhibition as well as laccase production by agar well diffusion method. Out of 91 cultures, #20CMBANEY and #20(b)VVLPM showed the potential tyrosinase inhibitory and laccase activity respectively. Further, Crude Dichloromethane extract of #20CMBANEY was resolved into 6 bands using Hexane: Toluene solvent system. In vitro anti microbial assay was carried out by agar well diffusion method in which methanol fraction of #20CMBANEY exhibited the maximum anti microbial activity against E.coli and ethyl acetate fraction against B.subtilis. In vitro anti-oxidant assay was carried out by using spectrophotometer, hexane fraction exhibits the maximum 63.2% of free radical scavenging activity.The potential endophytic fungus #20CMBANEY was identified as Nigrospora sp. by morphotaxonomy. ITS region of approximately 500bp was amplified and further speciation of fungi will be deduced after analyzing sequencing data of ITS region.