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Abstract: Aims: A new L‐asparaginase produced by Streptomyces ansochromogenes UFPEDA 3420 actinobacteria was used in this study against human lymphocyte cultures to evaluate the immunological profile induced by this enzyme. Methods and Results: Cultures of lymphocytes were stimulated with S. ansochromogenes L‐asparaginase, and cytotoxicity, cell viability, cell stimulation and cytokine production were analysed. This new S. ansochromogenes L‐asparaginase induced activation and proliferation of the TCD8+ lymphocyte subset and produced higher TNF‐α, IFN‐γ, IL‐2 and IL‐10 levels in a 24‐h assay. Conclusion: Streptomyces ansochromogenes L‐asparaginase is a promising molecule to be used in in vivo models and to deepen preclinical tests against acute lymphoblast leukaemia. Significance and Impact of Study: L‐asparaginase is an indispensable component of the chemotherapeutic treatment of acute lymphoblast leukaemia (ALL) and acute myeloid leukaemia (AML). Currently, drugs such as Asparaginase®, Kidrolase®, and Elspar® and Erwinase® are efficient against leukemic disease, but promote immunosuppression and other side effects in human organisms. Our purified S. ansochromogenes L‐asparaginase showed promissory results inducing, in vitro, higher immunostimulation in human PBMC, especially in T CD8+ lymphocyte subsets. [ABSTRACT FROM AUTHOR]