Abstract
Abstract Background The present study focused on utilization of agrowaste byproducts generated from oil mill for L-asparaginase enzyme production using Serratia marcescens under solid state fermentation. Classical and statistical methods were employed to optimize the process variables and the results were compared. Results The classical one factor at a time (OFAT) and response surface methodology (RSM) are employed to optimize the fermentation process. When used as the sole carbon source in SSF, coconut oil cake (COC) showed maximum enzyme production. The optimal values of substrate amount, initial moisture content, pH and temperature were found to be 6 g, 40%, 6 and 35°C respectively under classical optimization method with maximum enzyme activity of 3.87 (U gds-1). Maximum enzyme activity of 5.86 U gds-1 was obtained at the predicted optimal conditions of substrate amount 7.6 g of COC, initial moisture content of substrate 50%, temperature 35.5°C and pH 7.4. Validation results proved that a good relation existed between the experimental and the predicted model. Conclusions RSM optimization approach enhances the enzyme production to 33% when compared to classical method. Utilization of coconut oil cake as a low cost substrate in SSF for L-asparaginase production makes the process economical and also reduces the environmental pollution by converting the oil mill solid waste into a useful bioproduct.
Highlights
The present study focused on utilization of agrowaste byproducts generated from oil mill for L-asparaginase enzyme production using Serratia marcescens under solid state fermentation
Optimization of bioprocess by one factor at a time (OFAT) is a wellstudied method but it has some disadvantages like more time consumption, more number of experimental runs and lack of knowledge about the interaction between the variables involved in the process [26]
L-asparaginase activity was determined by measuring the amount of ammonia released by nesslerization according to the method described by Wriston and Yellin [28]. 0.2 mL of cell free supernatant was mixed with 0.8 mL of 0.1 M sodium borate buffer and 1 mL of 0.04 M L-asparagine and incubated for 10 min. 0.5 mL of 15% TCA was added to stop the reaction and again centrifuged for 10000 rpm for 10 minutes. 0.2 mL of the supernatant was taken in a test tube in which 3.6 mL distilled water and 0.2 mL Nessler’s reagent were added
Summary
The present study focused on utilization of agrowaste byproducts generated from oil mill for L-asparaginase enzyme production using Serratia marcescens under solid state fermentation. Because of the above disadvantages, Solid state fermentation (SSF) has emerged as an alternate cost- effective process for bioproducts production by utilizing the agrowastes and byproducts [19]. It has been employed for production of various microbial metabolites such as enzymes [3,4,20], organic acids [21,22,23] and antibiotics [24,25]. The main objective of this study was to produce L-asparaginase by selecting an appropriate low cost substrate and to determine the optimal level of the process variables by both OFAT and RSM methods
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