Partially Purified L-asparaginase with Low Glutaminase and Urease Co-Activities of Bacteria from the Rancabuaya Coast, Indonesia
Abstract
L-asparaginase, enzyme that can hydrolyses L-asparagine into L-aspartate and ammonia is used as a therapeutic enzyme in treating Acute Lymphoblastic Leukemia (ALL). The use of this enzyme is restricted by the presence of dual substrate specificity towards asparagine and glutamine, which causes side effects. The objectives of this research are to isolate and to identify marine bacteria from Rancabuaya Coast that generate L-asparaginase with low glutaminase and urease co-activity and to produce and to measure the asparaginase activity using the Nessler reagent. Bacteria were isolated from seawater, and screened using Zobell Marine media containing either L-asparagine, glutamine or urea, with phenol red as an indicator. The bacterial isolate with the highest asparaginase activity and relatively lower glutaminase and urease activity, was identified by 16S rRNA gene sequence. This bacterial isolate, RB3, was identified as Pseudoalteromonas tetraodonis GFC with greater than 99 % homology. Enzyme specific activity of crude extracellular and intracellular enzyme were 72.30 U mg-1 and 67.18 U mg-1 respectively, while the highest enzyme specific activity from ammonium sulphate fractionation was found at 40-60% saturation (F3), which was 136.03 U mg-1. SDS-PAGE of the enzyme solutions showed the presence of a 35 kDa band suspected to be the L-asparaginase protein.
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