Pongamia pinnata (Millettia pinnata L.) Pierre commonly known as Karanj or Karanja in Hindi. In the present time Pongamia is used for production of biodiesel. Pongamia oil is used as biodiesel. Biodiesel refers to a non-petroleum-based diesel fuel consisting of long-chain alkyl (methyl, propyl ethyl) esters. Biomolecules such as proteins and enzymes are responsible for the functions and characters of the plant. In the present study protein profiling and enzyme estimation of different accessions of Pongamia has been done to study the genetic variability. As we know, proteins and enzymes play an important role in different functions and characters of the plant. The present study was carried out in the Tree Improvement Lab of NRCAF, Jhansi during the months of January 2009 to June 2009. To study the protein content and enzyme content of 18 accessions of Pongamia pinnata L., samples were collected from NRCAF, Experimental Field. The research work on “Study on biochemical activities in different accessions of Pongamia pinnata (L.)” is still at an infancy stage. Here it has been tried to accumulate as much information as available on Pongamia pinnata (L.) in relation to present study.