Sequential Optimization Methods for Augmentation of Marine Enzymes Production in Solid-State Fermentation: l-Glutaminase Production a Case Study.

There is an increased l-glutaminase market worldwide due to its relevant industrial applications. Salt tolerance l-glutaminases play a vital role in the increase of flavor of different types of foods like soya sauce and tofu. This chapter is presenting the economically viable l-glutaminases production in solid-state fermentation (SSF) by Aspergillus flavus MTCC 9972 as a case study. The enzyme production was improved following a three step optimization process. Initially mixture design (MD) (augmented simplex lattice design) was employed to optimize the solid substrate mixture. Such solid substrate mixture consisted of 59:41 of wheat bran and Bengal gram husk has given higher amounts of l-glutaminase. Glucose and l-glutamine were screened as a finest additional carbon and nitrogen sources for l-glutaminase production with help of Plackett-Burman Design (PBD). l-Glutamine also acting as a nitrogen source as well as inducer for secretion of l-glutaminase from A. flavus MTCC 9972. In the final step of optimization various environmental and nutritive parameters such as pH, temperature, moisture content, inoculum concentration, glucose, and l-glutamine levels were optimized through the use of hybrid feed forward neural networks (FFNNs) and genetic algorithm (GA). Through sequential optimization methods MD-PBD-FFNN-GA, the l-glutaminase production in SSF could be improved by 2.7-fold (453-1690U/g).