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Betulin (B) and Betulinic acid (BA) are naturally occurring lupane-type pentacyclic triterpenes with significant pharmacological relevance like anti-tumor and anti-HIV activities. AIDS is a mixture of conditions that gradually reduces the immunity of the host making them more susceptible to even common infections like influenza. The cure of HIV is generally based on a blend of 3 or more type of medication which is named as HAART (Highly active anti Retroviral Therapy). This blending of medication supports in the reduction of chances for virus replication by masking their mutations. Even though there are more than 25 known drugs with diverse mechanism of action, available in the market but still there is need of alternative mechanism of action with better cure as well lower side effect. Now Betulin and Betulinic acid (BA) with its derivatives represents as significant group of anti-HIV agents with novel mechanisms. Previously Betulin and Betulinic acid derivatives were chemically synthesized by altering different groups specially at C3, C20 and C28 positions but here our aim is through microbial transformation. Range of betulin 0.25 to 4.0 mg/l were tried and 1.4 mg/l was found best for maximum growth at 250 ml shake flask. Highest growth 58.51mg/ml and yield of betulinic acid 22.32 mg/l (statistically optimization). Biotransformation was confirmed by RP-HPLC analysis and further characterization of transformed product as betulinic acid was done by ESI-MS.