Optimization of process variables for enhancement of L-asparaginase with reduced glutaminase productivity by statistical approach from Pectobacterium carotovorum MTCC 1428

摘要

Aim. The aim of this paper was to optimize the process variables for the enhanced production of L-asparaginase having reduced glutaminase activity from Pectobacterium carotovorum by statistical methods. Methods. Various statistical methods like Taguchi and TSM were applied to enhance the productivity of L-asparaginase from Pectobacterium carotovorum MTCC 1428. Crude extract of Pectobacterium is analyzed for L-asparaginase and L-glutaminase activities and enzyme kinetics was determined. Results. L-asparaginase productivity was enhanced when 0.1% asparagine and 5% galactose were supplemented to glucose yeast extract medium. Along with increase in asparaginase activity, 26% of glutaminase activity was also noticed. Media containing 1% galactose and 1% maltose along with 0.1% asparagine showed maximum L-asparaginase with relatively less or negligible glutaminase activity. Statistical experimental designs like Taguchi and CCD of RSM were applied to further fine tune the media and enhanced the L-asparaginase yield from 5 to 24.33 U/mg. Conclusion. Pectobacterium carotovorum MTCC 1428 could be a better source for the production of L-asparaginase and also it is to declare that the formulated media composition with galactose as carbon source has enhanced the enzyme production with reduced glutaminase side activity for better treatment of ALL.