Experimental design applied to the optimization and partial characterization of pectin-methyl-esterase from a newly isolated Penicillium brasilianum

Abstract

The objective of this work was to optimize the medium composition for maximum pectin-methylesterase (PME) production from a newly isolated strain of Penicillium brasilianum by submerged fermentation. A Plackett-Burman design was first used for the screening of most important factors, followed by a 2³ full experimental design, to maximize the enzyme production. The maximum pectinmethyl-esterase activity was 4.0 U/mL at 24 h of bioproduction using a pectin concentration of 32.0 g/L, yeast extract of 30.0 g/L, potassium phosphate of 8.0 g/L, iron (II) sulfate of 0.02 g/L, at 30°C, stirring rate of 180 rpm and initial pH of 5.5. The kinetic evaluation showed that after 27 h of fermentation, a consumption of 15% of total organic carbon and 10% of nitrogen was observed. The crude enzymatic extract kept about 80% of its initial activity after 1848 h under low temperatures. An increase of PME activity was observed after incubation at high temperatures. The residual activity of the extracts after 1728 h of incubation was about 95% for all tested pH values (5, 7, 9 and 11). The application of 0.5% (v/v) of PME crude extract for clarification of peach juice showed a reduction on the viscosity (7.20%) and turbidity (14.11%).

Keywords: Penicillium brasilianum, pectin-methyl-esterase, experimental design, pectin-methyl-esterase (PME).

African Journal of Biotechnology Vol. 12(40), pp. 5886-5896