PRODUCTION AND CHARACTERIZATION OF INULINASE BY SOLID STATE FERMENTATION IN
BATCH AND FED-BATCH MODE
V. Astolfi1, H. Treichel2, M. A. Mazutti3, E. Rigo, M. L. Ugalde4. 1 Department of Food Engineering – URI,
Campus Erechim, Av. Sete de Setembro, 1621 - CEP: 99700-000 - Erechim - RS - Brazil, Tel.: (54) 35209000 - Fax: (54) 3520-9090. 2 Laboratory of Biochemical Engineering School of Chemistry and Food,
Federal University of Rio Grande (FURG) - P.O. Box 474 - CEP: 96201-900 - Rio Grande - RS - Brazil. 3
Department of Chemical Engineering – Federal University of Santa Maria, Av. Roraima, 1000, Santa Maria RS, P.O. Box 97105-900 - Santa Maria - RS - Brazil. 4 Federal Institute Farroupilha, Campus Júlio de
Castilhos, P.O. Box 38 CEP 98130-000, Júlio de Castilhos - RS, Brazil. [email protected]
(55)3271-9500.
Inulinases are important enzymes for the production of fructose by the enzymatic hydrolysis of inulin and in
the production of fructooligosaccharides, a functional ingredient for food industries, considered a prebiotic
compound. The objective of this work was the production and characterization of inulinases by solid state
fermentation using strategies of batch and fed-batch fermentation using the yeast Kluyveromyces
marxianus NRRL Y-7571. To evaluate the kinetics of production of the inulinase, fermentations were
performed destructive, i.e., each measured time interval corresponding to a new fermentation. The partial
characterization of the crude enzymatic extract obtained by fed-batch fermentation was also carried out.
Among the results obtained for inulinase production showed that the best experimental condition was by fedbatch fermentation, resulting in a maximum production of 470.72 U.gds. -1, after 24 hours of fermentation.
The enzymatic extract obtained by fed-batch mode was characterized and presented optimum temperature
and pH in the range of 52-57 ºC and 4.8-5.2, respectively. For a temperature range from 40 to 70 ºC the
enzyme presented decimal reduction time, D-value, ranging from 5748 to 47 h, respectively. For a pH range
from 3.5 to 5.5 the enzyme showed good stability, presenting D-values higher than 2622 h. In terms of
Michaelis-Mentem parameters were demonstrated that the crude inulinase activity presented higher affinity
for substrate sucrose compared to inulin.