Parameters optimization for enzymatic assays using experimental design

• Main Authors: J. F. M. Burkert, S. J. Kalil, F. Maugeri Filho, M. I. Rodrigues
• Format: Article
• Language: English
• Published: Brazilian Society of Chemical Engineering 2006-06-01
• Series: Brazilian Journal of Chemical Engineering
• Subjects: inulinase; Kluyveromyces marxianus; activity; factorial design and response surface analysis
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66322006000200002

The conditions for maximization enzymatic activity were determined using experimental design and inulinase from Kluyveromyces marxianus ATCC 16045. The effects of substrate concentration (sucrose and inulin), pH and temperature on inulinase activity were verified using four factorial design and surface response analysis. Using sucrose as substrate. It has bean shown that the effects sucrose on enzymatic activity is not statistically significant and the best condition for the highest activity (110 U/mL) was achieved with temperature between 60°C and 68°C and pH between 4.5 and 5.0. Using inulin as substrate it was verified that temperature is the only variable statistically significant and the maximum activity was 7.3 U/mL at temperature between 50°C and 51°C.