Center of Technology, School of Chemistry, Laboratories of Bioprocess Development, Federal University of Rio de Janeiro, 21949-900, Rio de Janeiro, Brazil.
Appl Biochem Biotechnol. 2010 May;161(1-8):411-22. doi: 10.1007/s12010-009-8875-6. Epub 2009 Dec 15.
The present study aimed at maximizing cellulase production by Penicillium funiculosum using sequential experimental design methodology for optimizing the concentrations of nitrogen sources. Three sequential experimental designs were performed. The first and the second series of experiments consisted of a 2(4) and a 2(3) factorial designs, respectively, and in the third one, a central composite rotational design was used for better visualizing the optimum conditions. The following nitrogen sources were evaluated: urea, ammonium sulfate, peptone, and yeast extract. Peptone and ammonium sulfate were removed from the medium optimization since they did not present significant statistical effect on cellulase production. The optimal concentrations of urea and yeast extract predicted by the model were 0.97 and 0.36 g/L, respectively, which were validated experimentally. By the use of the desirability function, it was possible to maximize the three main enzyme activities simultaneously, which resulted in values for FPase of 227 U/L, for CMCase of 6,917 U/L, and for beta-glucosidase of 1,375 U/L. These values corresponded to increases of 3.3-, 3.2-, and 6.7-folds, respectively, when compared to those obtained in the first experimental design. The results showed that the use of sequential experimental designs associated to the use of the desirability function can be used satisfactorily to maximize cellulase production by P. funiculosum.
本研究旨在通过青霉(Penicillium funiculosum)的序贯实验设计方法来最大化纤维素酶的生产,以优化氮源浓度。进行了三个序贯实验设计。第一和第二系列实验分别为 2(4)和 2(3)析因设计,而在第三个实验中,使用中心复合旋转设计可以更好地观察最佳条件。评估了以下氮源:尿素、硫酸铵、蛋白胨和酵母提取物。由于对纤维素酶生产没有显著的统计学影响,因此从培养基优化中去除了蛋白胨和硫酸铵。模型预测的尿素和酵母提取物的最佳浓度分别为 0.97 和 0.36 g/L,实验验证了这一预测。通过使用理想函数,可以同时最大化三种主要酶活性,从而使 FPase 的值达到 227 U/L、CMCase 的值达到 6,917 U/L 和β-葡萄糖苷酶的值达到 1,375 U/L。与第一个实验设计相比,这些值分别增加了 3.3 倍、3.2 倍和 6.7 倍。结果表明,使用序贯实验设计并结合理想函数的使用,可以令人满意地最大化青霉(Penicillium funiculosum)的纤维素酶生产。