Abstract
Response surface methodology was used to study the cumulative effect of the nutritional parameters and to enhance the production of extracellular α-amylase in solid-state fermentation by Thermomyces lanuginosus ATCC 58157. These nutritional parameters considered include carbon source (soluble starch), nitrogen source (peptone) and a concentrated mineral medium. For obtaining the mutual interaction between the variables and optimizing these variables, a 23 factorial central composite design using response surface methodology was employed. The optimal calculated values of tested variables for maximal production of α-amylase were: soluble starch, 71.10 g/Kg; peptone, 91.97 g/Kg and mineral salts solution, 175.05 ml/Kg with a predicted α-amylase activity of 5.085 ´ 105 U/Kg of wheat bran. These predicted optimal parameters were tested in the laboratory and the final α-amylase activity obtained, 4.946 ´ 105 U/Kg of wheat bran, was very close to the predicted value. Key words: α-Amylase, Thermomyces lanuginosus, solid state fermentation, optimization, response surface methodology.
Highlights
-Amylase have most widely been reported to occur in microorganisms, they are found in plants and animals. -Amylase are extracellular enzymes that randomly cleave the 1,4-α-D-glucosidic linkages between adjacent glucose units in the linear amylose chain
Since thermostability is a feature of most of the enzymes sold in bulk for industrial application, thermophilic microorganisms are of special interest for the production of thermophilic amylases
T. lanuginosus strain ATCC 58157 was maintained on potato dextrose agar (PDA) at 4°C and transferred every 6-7 weeks
Summary
-Amylase have most widely been reported to occur in microorganisms, they are found in plants and animals. -Amylase (endo-1,4-alpha-D-glucan glucohydrolase, EC 3.2.1.1) are extracellular enzymes that randomly cleave the 1,4-α-D-glucosidic linkages between adjacent glucose units in the linear amylose chain. The enzymes are reported to be thermostable, with the half-life of amylase and glucoamylase being 0.6 and 4 h, respectively at 70°C Due to their increased thermostability, these enzymes are potentially useful in the starch industry for production of maltose and glucose. The classical method for the optimization of medium and cultural conditions involves one variable at a time, while keeping the other parameters at fixed levels This method is generally time consuming and requires a considerable number of experiments to be carried out and does not include interactive effects among the variables (Dey et al, 2001). The aim of the work includes the statistical optimization of supplementary nutrients predicted to play a significant role in enhancing -amylase production in solid-state fermentation
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