METHODS AND RESULTS: Five pathogen: antagonist inoculum ratios (100:0, 75:25, 50:50, 25:75, and 0:100) were employed. Non-aflatoxigenic Aspergillus flavus Af1KD and Af5TD, and Penicillium janthinellum were used against aflatoxin B1 (AFB1) and aflatoxin B2 (AFB2) by A. flavus. Against OTA by A. niger, and fumonisin B1 (FB1) and fumonisin B2 (FB2) by Fusarium verticillioides and F. proliferatum, respectively, the antagonists Trichoderma asperelloides, T. asperellum, and T. harzianum were used. Non-aflatoxigenic A. flavus Af1KD was the most effective against AFB1 and AFB2 at all tested ratios and substrates. All Trichoderma spp. were effective against OTA by A. niger on grain corn agar at all tested ratios. Trichoderma asperelloides and T. asperellum were effective against FB1 and FB2 produced by F. verticillioides at all tested ratios and substrates. Trichoderma asperelloides was effective against FB1 and FB2 by F. proliferatum at all tested ratios and substrates.
CONCLUSION: The native biocontrol candidates were effective against mycotoxigenic fungi and mycotoxin production on grain corn agar and grain corn kernels, and could be developed into biocontrol agents.
RESULTS: The production medium was first optimized using a statistical optimization approach to increase pectinase production. A maximal enzyme concentration of 76.35 U/mL (a 2.8-fold increase compared with the initial medium) was produced in a medium composed of (g/L): pectin, 32.22; (NH4)2SO4, 4.33; K2HPO4, 1.36; MgSO4.5H2O, 0.05; KCl, 0.05; and FeSO4.5H2O, 0.10. The cultivations were then carried out in a 16-L stirred tank bioreactor in both batch and fed-batch modes to improve enzyme production, which is an important step for bioprocess industrialization. Controlling the pH at 5.5 during cultivation yielded a pectinase production of 109.63 U/mL, which was about 10% higher than the uncontrolled pH culture. Furthermore, fed-batch cultivation using sucrose as a feeding substrate with a rate of 2 g/L/h increased the enzyme production up to 450 U/mL after 126 h.
CONCLUSIONS: Statistical medium optimization improved volumetric pectinase productivity by about 2.8 folds. Scaling-up the production process in 16-L semi-industrial stirred tank bioreactor under controlled pH further enhanced pectinase production by about 4-folds. Finally, bioreactor fed-batch cultivation using constant carbon source feeding increased maximal volumetric enzyme production by about 16.5-folds from the initial starting conditions.