Assessment of Optimal Conditions for L-asparaginase Production by Endophytic Lasiodiplodia Pseudothreobromae

Abstract

L-Asparaginase (LA) (E.C. 3.5.1.1) is a naturally occurring enzyme initiate in diverse microbes. LA catalyzes L-asparagine amidohydrolase an essential amino acid for leukemic cells, into aspartate and ammonia. L-asparaginase is the first line therapeutic enzyme that has been abundantly studied by researchers due to its antitumor properties. In Biopharmaceutical, L-asparaginase has been used in the treatment of acute lymphoblastic leukemia (ALL), acute myeloblastic leukemia (AML), and other lymphoid melanomas, in combination with drugs. In the medical field, endophytic bioactive compounds are playing a major role in the modern new medication because of their wide variety of biological activities as antibiotic, anticancer, antioxidant, and anti-inflammatory agents. Previous study suggested that the source of L-asparaginase supplied by recombinant enzymes from Escherichia coli, Streptomyces albidoflavus and Erwinia chrysanthemi. However, from bacterial sources proteins associated to immunogenicity problems such as anaphylaxis, hypersensitivity Type IV, hepatotoxicity, leukopenia, pancreatitis, thrombosis, neurological crises and which leads to the search for a better enzyme source. Thus, the use of endophytic fungi as substitute source which comes in trend. These fungi have antimicrobial activity and hence have a great role as biocontrol agent. The current study suggested that the L-asparaginase production and biochemical characterization pointing to increase the LA production. In the current investigation, endophytic fungi Lasiodiplodia pseudotheobromae isolated from Aegle marmelos was cultured in our laboratory and used to determine the potential of L-asparaginase production by both qualitative and quantitative assays. By screening LA production with the enzymatic zone of activity showed with the value of 0.56. The L-asparaginase activity of the crude protein showed in the range at 5.18 U/min/ml. The optimum temperature (25±2°C) and pH (7) were showed the maximum enzymatic zone of activity at 0.50 and 0.51 respectively. The maximum enzymatic substrate concentration shown at 0.9% (w/v).