Experimental Design Applied to the Optimization and Partial Characterization of Pectin Liase from a Newly Isolated Penicillium brasilianum

Abstract

Penicillium brasilianum was previously isolated from tea and identified by molecular biology technique. A PlackettBurman design, followed by a complete second order design was used for the screening of most important factors and to maximize the pectin liase (PMGL) activity, respectively. The maximum PMGL activity by P. brasilianum achieved was 9.0 U/mL after 48 h of cultivation in a medium containing pectin (33.0 g/L), yeast extract (30.0 g/L) and potassium phosphate (2.0 g/L) at 30ºC, with a stirring rate of 180 rpm, initial pH 5.5 and 5x106 spores/mL inoculum size. The kinetic evaluation in terms of substrate consumption demonstrated that the maximum production of PMGL was at 72 h, and 40% of the total organic carbon, 25% of the nitrogen, 88% of the magnesium, 13% of the potassium and 66% of the iron were consumed. The pH remained almost stable during the whole period of production (5.33 to 4.9). The partial characterization of the crude PMGL enzyme extract showed optimal pH and temperature of 5.5 and 37°C, respectively.

Biochemical characterization and kinetic/thermodynamic study of Aspergillus tamarii URM4634 β‐fructofuranosidase with transfructosylating activity

“…To investigate the possible effect of temperature and pH interaction on FFase activities and identify the optimum conditions for both, we performed a set of cultivations according to a CCRD, where the levels of these two independent variables were selected based on the above individually determined optimum values. Such an approach, which is still poorly reported in the literature for biochemical characterization, provides valuable information for a better understanding of how operating variables influence the catalytic process and was already used to investigate the biochemical profile of enzymes of industrial interest such as proteases, cellulases, pectin lyases, and FFase …”

“…The optimum pH was 6.0 for both activities (Figure 1b), a value coincident with that reported for an A. oryzae FTase. 31 such as proteases, 32 cellulases, 33 pectin lyases, 34 and FFase. 35 From the matrix of independent variables and responses used in CCRD (Table 1), one can see that the highest hydrolytic (159.81-160.63 U mL −1 ) and transfructosylating (63.16 U mL −1 ) activities were obtained in the central points (runs 9 and 10 carried out at 50 C and pH 6.00) and in run 6 (50 C and pH 7.41), respectively, conditions under which the response surfaces showed optimum regions for both ( Figure 2).…”

Insights into Penicillium brasilianum Secondary Metabolism and Its Biotechnological Potential

“…Penicillium is one of the most used genera in biotechnology and Penicillium brasilianum is also a very important biotechnological target for enzyme production as reviewed here [ 15 , 16 , 17 ]. Jørgensen et al [ 15 ] described a series of biotechnological enzymes produced by an isolate of P. brasilianum IBT 20888 isolated from seaweed in Denmark.…”

“…Zeni et al [ 16 ] reported pectinase-producing microorganisms with polygalacturonase activity including the isolated Penicillium sp. from tea, which was posteriorly identified as Penicillium brasilianum by molecular biology techniques [ 17 ].…”

“…Zeni et al [ 17 ] described the optimization and partial characterization of a pectin liase from P. brasilianum isolated from tea. Based on factorial experiments and Plackett-Burman design, the authors were able to verify the best conditions for highest pectin lyase (PMGL) activity.…”