New Procedures for Purification of l-Asparaginase with High Yield from Escherichia coli

AUTOR(ES)

Roberts, Joseph

RESUMO

l-Asparaginase is now known to be a potent antineoplastic agent in animals and has given complete remission in some human leukemias. Extensive clinical trials of this enzyme, however, were not possible in the past because of inadequate production of this substance. We have developed practical procedures for producing l-asparaginase in yields of sufficient quantity and purity for more extensive clinical evaluation. The nutritional requirements for optimal production of biologically active l-asparaginase by a strain of Escherichia coli have been ascertained. The highest yields of enzyme were obtained when cells were grown aerobically in a corn steep medium. Good enzyme production was associated with media containing l-glutamic acid, l-methionine, and lactic acid. The addition of glucose to the medium, however, resulted in depressed production of l-asparaginase. Sodium ion appeared to suppress l-asparaginase production. With the procedure described for isolation of biologically active l-asparaginase from E. coli, stable l-asparaginase preparations with a specific activity of 620 IU per mg of protein (1,240-fold purification with 40% total recovery) were obtained.

Documentos Relacionados

• Production of l-Asparaginase II by Escherichia coli
• Purification and Properties of l-Asparaginase from Serratia marcescens1
• Production, purification and characterization of L-asparaginase from Streptomyces gulbargensis
• l-Asparaginase from Proteus vulgaris1
• Expression, purification and crystallization of Helicobacter pylori l-asparaginase