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Appl Environ Microbiol. 1969 October; 18(4): 550-554
Copyright © 1969 American Society for Microbiology. All Rights Reserved.

Production of Tumor-Inhibitory L-Asparaginase by Submerged Growth of Serratia marcescens¹

^a Research Division, Bristol Laboratories Division of Bristol-Meyers, Syracuse, New York 13201

ABSTRACT

Production of a tumor-inhibitory asparaginase by submerged fermentation with Serratia marcescens ATCC 60 was studied to ascertain optimal nutritional conditions for large-scale production leading to enzyme purification studies. Five strains of S. marcescens were screened in shake-flask studies and were found to produce 0.8 to 3.7 IU/ml 48 hr after inoculation. The requirements for asparaginase production with S. marcescens ATCC 60, the high producing strain, included the following: 4% autolyzed yeast extract medium (initial pH 5.0), an incubation temperature of 26 C, and limited aeration for a zero level of dissolved oxygen during the fermentation. Addition of various carbohydrates to the fermentation medium did not enhance yields. The peak cell population in the fermentation medium and the maximal asparaginase yields occurred simultaneously. Highest enzyme yields were found when the pH of the fermentation cycle rose to approximately 8.5. Yields of 4 IU of asparaginase/ml of cell suspension have been obtained consistently in 40 to 42 hr from 10-liter volumes (500 ml/4-liter bottle) produced on a reciprocating shaker. Scale-up to a 60-liter fermentor yielded 3.1 IU/ml in 35 hr.

¹ Presented in part at the 69th Annual Meeting of the American Society for Microbiology, Miami Beach, Fla., 4-9 May 1969.