Microbial Transformation Of Ethyl Acetate Fraction Of Alpinia galanga Extract By Aspergillus niger, Aspergillus oryzae, Rhizopus oligosporus, Penicillium sp, and Saccharomycesrouxii And Cytotoxic Assay Of Its Produced Metabolites On T47D Cell Line

Cancer is quickly becoming one of the first leading cause of death worldwide, and one type of cancer with the highest mortality rate in women is breast cancer. Lengkuas (Alpinia galanga) is one of the plants known among others to have cytotoxic activity against cancer cells. The biotransformation process has gained importance over chemical technologies because of process controls, manipulations of microorganisms, safety, and reproducibility. This study aimed to determine whether the use of biotransformation of ethyl acetate of A.galanga extract within fungi will lead to a new metabolite and to determine its cytotoxic activity effect against T47D cells. The simplisia of plants is macerated with ethanol 96 % and then fractionated using hexane:ethyl acetate (3:1) and choroform:methanol (9:1) then TLC test was performed using a UV spectrophotometer at λ=254nm and λ=366nm. The method used in the biotransformation process was, a sub-culture of each fungus into a solid medium culture potato dextrose agar (PDA) plate and pre-incubate at 28° C for 7 days. Then, was transferred to a liquid medium culture. The identification and purification of the new metabolites were carried out with TLC and LC-MS. The TLC profile after biotransformation did not show any new spots within the treatment sample (Aspergillus.niger, Aspergillus.oryzae, Penicillium sp) except the treatment sample (Saccharomyces rouxii, Rhizopus oligoropus) where there is no presence of a spot. The LC-MS profile shown the same base peak of A.galanga extract (234.84) within each treatment sample (Aspergillus.niger, Aspergillus.oryzae, Saccharomyces rouxii) except the treatment sample (Penicillium sp, Rhizopus oligoropus) where there is no record of the base peak of A.galanga extract. The result showed that they were no change in the chemical content of the compound of A.galanga extract and the treatment samples. The method used in the cytotoxic test was the MTT assay. Doxorubicin was used as a control positive. The IC50 obtained from A.galanga extract sample was 55.55 µg/mL, while in the other samples (A.galanga extract+ fungi) there was no activity. This research concludes that ethyl acetate fraction of A.galanga extract biotransformation did not lead to a new metabolite, nor have no cytotoxic activity effect against T47D cells. Keywords: Biotransformation, sub-culture, fermentation, fungi, Alpinia galanga extract, TLC, LC-MS, MTT, and T47D