The entomopathogenic fungus Metarhizium anisopliae B is a powerful biological control agent against Monochamusalternatus, a crucial mediator of the pinewood nematode Bursaphelenchus xylophilus. In this study, production of destruxins(dtxs), insecticidal cyclic hexadepsipeptides, was monitored in the submerged culture of M. anisopliae B. Three typesof dtxs, i.e., destruxin A, B, and E, were produced during the culture. Among the three dtxs, the production yield ofdestruxin A was best, followed by destruxin B and E. Destruxin A production was increased when pH was controlledat 6.0, whereas production of destruxin E was not affected by the pH control. The highest yield of dtxs A, B, and Ewere 16.4, 7.3, and 6.1 mg L-1, respectively. Considering that process for dtxs production has not been optimized, M.anisopliae B has more powerful implication as a biocontrol agent.

The entomopathogenic fungus Metarhizium anisopliae is one of potent biological control agents against a variety ofinsect pests. In this study, we investigated enzyme production of M. anisopliae strains A and B. They produced extracellularenzymes for degrading the epidermis of Monochamus alternatus, a crucial mediator of the pinewood nematode Bursaphelenchusxylophilus. With Q-TOF MS/MS analysis, 29 kDa protein, a major band on a SDS-PAGE gel, was identified as subtilisin-likeserine protease PR1A. M. anisopliae A produced an extracellular enzyme more efficiently than M. anisopliae B: however,enzyme activities targeted for the cuticle were comparable. Our results suggest that the two strains of M. anisopliae havethe biological potential against M. alternatus with insecticidal protease production.

Oxalic acid has a nematicidal activity against the root-knot nematode Meloidogyne incognita. High producer of oxalic acid was isolated, and then named as Aspergillus niger F22. Oxalic acid production was investigated under various temperatures from 20 – 33oC and rotational speeds in 5 L jar fermenters. Yield of oxalic acid increased with decreasing temperature. The highest yield was obtained at 23oC, showing the yield of oxalic acid of 8.7 g/L, whereas oxalic acid production was least at 33oC. At 20oC, the yield was lower than that of 23oC. At a rotational speed of 300 rpm, serious oxygen depletion was present from 48 - 72 h, resulting in low productivity of 26.2 mg /L·h. When a rotational speed was set at 600 rpm, dissolved oxygen tension was over 40% and oxalic acid production increased up to approximately 55%. Viscosity during the culture differed with temperatures. Viscosity increased with the increment of temperatures. When A. niger F22 was cultured at 23oC, viscosity was 810 cP, which was favorable for oxalic acid production.

Biological control has been tried for integrated pest management. It is often comparable, safe, and environment-friendly, making itself an alternative for chemical agents. Filamentous microorganisms, i.e., fungi and streptomystes, produce many kinds of useful metabolites, and some of them have been developed as a biocontrol agent. However, they still have a long way because of the concern of manufacturing cost. Therefore, process development was intensively studied to meet cost-effectiveness. Operating conditions of bioreactor, e.g., agitation and aeration, had an effect on biological and physiological responses such as mycelial morphology, oxygen and nutrient transfer. Understanding relationship between operating parameters and microbial responses in terms of growth, substrate and oxygen consumption, and production yield was critical for process development. This study dedicated to build strategies for mass production of biological control agent using aerobic filamentous microorganisms.