Application of Plackett-Burman design for the optimization of protease production by Aspergillus niger

Sana Ghorri, Ouided Benslama, O. Benserradj, Ilhem Mihoubi
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Abstract

Proteolytic enzymes represent one of the largest groups of industrial enzymes. Fungal proteases have been used for the large-scale production of industrial enzymes. The optimization of enzyme production is a very important objective, where the aim is to further improve the production process. Therefore, the application of an experimental statistical method is essential. In this study, the production of the enzyme protease was carried out by fermentation of a mold (Aspergillus niger) using two media, one based on wheat bran and the other based on peas chick. The optimization of the synthesis of the enzyme was carried out using a statistical method of experimental planning based on the Plackett and Burman matrices. The variables used are the 3 production factors temperature, pH, and substrate concentration. The results were modeled according to multiple linear regression. The results obtained revealed that the strain A.niger gave an excellent activity reaching 740 IU for the wheat bran medium and 194.54 IU for the chickpea medium. The study of pH optima and incubation temperature has shown that the protease produced by A. niger has an optimum pH equal to 6 and an optimum temperature of 40°C for wheat bran medium and has an optimum pH equal to 5 and an optimum temperature of 30°C for chickpeas. The results obtained from the experimental designs of Plackett and Burman proved to be ideal for the selection of factors influencing protease production.
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应用Plackett-Burman设计优化黑曲霉生产蛋白酶
蛋白水解酶是工业酶中最大的一类。真菌蛋白酶已被用于大规模生产工业酶。酶生产的优化是一个非常重要的目标,其目的是进一步改善生产过程。因此,实验统计方法的应用是必不可少的。在本研究中,采用两种培养基,一种是麦麸培养基,另一种是豌豆鸡培养基,对一种霉菌(黑曲霉)进行了发酵生产蛋白酶。采用基于Plackett和Burman矩阵的实验规划统计方法对酶的合成进行优化。使用的变量是3个生产因素温度、pH和底物浓度。采用多元线性回归对结果进行建模。结果表明,该菌株对麦麸培养基和鹰嘴豆培养基的活性分别达到740 IU和194.54 IU。最适pH值和孵育温度的研究表明,黑曲霉产生的蛋白酶在麦麸培养基中最适pH值为6,最适温度为40℃;鹰嘴豆培养基中最适pH值为5,最适温度为30℃。从Plackett和Burman的实验设计中得到的结果证明是理想的选择影响蛋白酶生产的因素。
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