Screening for Type II L-Asparaginases: Lessons from the Genus Halomonas

Among the two types of bacterial L-asparaginases, only type II enzymes have been used in the treatment of acute lymphoblastic leukemia owing to their higher affinity for L-asparagine. However, current screening media used for the isolation of L-asparaginase-producing microorganisms do not discriminate between the two types of L-asparaginase. During an optimization study conducted to increase L-asparaginase production by environmental Halomonas isolates, it was noticed that the pattern of L-asparaginase production in response to variations in glucose concentration varied between different isolates suggesting that they differ in their ability to produce type II L-asparaginases, an observation that was confirmed by further experiments. Bioinformatics analysis of available Halomonas whole genome sequences revealed that indeed some species of this genus possess both L-asparaginase types while others possess only type I enzymes. By comparing the growth pattern of these isolates on different media, we propose that by omitting glucose, reducing the concentration of L-asparagine and providing an alternative nitrogen source in L-asparaginase screening media it may be possible to differentiate between type I and type II activities.