Therapeutic recombinant L-asparaginases: a comprehensive review and way forward.

L-Asparaginase (E.C.3.5.1.1; L-ASNase) hydrolyzes L-asparagine (an essential amino acid for the growth of leukemic cells) to aspartic acid and ammonia. It is obtained from various sources, including: bacteria, yeast, fungi, plants, and animals. It is used as a chemotherapeutic agent to treat acute lymphoblastic leukemia (ALL) and reduce acrylamide formation in baked and fried foods. Globally, various recombinant and pegylated L-ASNases formulations, including Escherichia coli and Erwinia-derived variants, are in Phase II or active clinical trials for ALL and related conditions across the USA, Brazil, and Canada. Some drugs are recruiting, and other variants of E. coli and E. chrysanthemi L-ASNases are already approved for marketing. Although L-ASNase is used as a therapeutic agent, immunogenic reactions and other adverse effects continue to limit its use. To enhance yield, L-ASNase isoforms are cloned and expressed in host cells, generating recombinants with distinct physicochemical properties and kinetic parameters. The enzymes' temperature and pH ranges vary from 25 to 100 °C and 6 to 10, respectively. Nowadays, enzymatic modifications, such as immobilization (chemical, physical, and PEGylation), mutagenesis, and PASylation, are used to overcome the limitations of commercialized L-ASNase-based drugs. Therefore, this review is a timely effort to compile and analyze the properties of recombinant L-ASNases and the contemporary techniques used to improve L-ASNase. A comprehensive study would help us better understand the kinetic parameters, biochemical properties, and modification trends of L-ASNase, enabling the development of robust, reliable therapeutics in the future.