Characterization of earthworm α-amylases for dietary supplement development and biomass utilization.

Earthworms are useful soil-decomposing animals that possess various saccharification enzymes such as cellulases and amylases. Earthworms have also been traditionally used as antipyretic agents and medicines for preventing thrombotic diseases such as brain infarction. We previously developed a novel earthworm dietary supplement with fibrinolytic, cellulase, and amylase activities using high-pressure technology. However, the optimal temperature and pH required for amylase activity in bioindustry have not yet been investigated. In the present study, we purified and characterized two α-amylases of Eisenia fetida Waki, EfAMY1 and EfAMY2, which were monomeric enzymes of 63.8 kDa and 64.0 kDa, with specific activities of 69.2 and 40.4 units/mg, respectively. The optimal pH was 5.5 for both enzymes, and the optimal temperatures were 45 °C and 35 °C for EfAMY1 and EfAMY2, respectively; however, the enzymes were stable over a wide pH range (5-10) and at high temperature (up to 40 °C). These amylases showed higher specific activity and cold tolerance than those previously reported. These data should help to promote the development of E. fetida AMYs as functional dietary supplements and in biomass utilization.

Characterization of two endoglucanases for the classification of the earthworm, Eisenia fetida Waki.

Eisenia fetida and Eisenia andrei are vermicomposting species that are used as model animals for testing chemical material toxicology. Eisenia spp. are grown commercially in various fields in Japan. However, these two species have not been classified because it is difficult to distinguish them morphologically; thus, all bred earthworms are called E. fetida. However, it has been proposed that these two species have different expression regulation mechanisms. Here, we classified a sample of earthworms purchased from several farms, confirming that both E. fetida and E. andrei are present in Japanese earthworm breeding programs. We also characterized two highly active endoglucanases (EfEG1 and EfEG2) from the E. fetida Waki strain, which contained strong fibrinolytic enzymes for improving human health. We confirmed that EfEG1 is 1371 bp long and belongs to GHF9. Thus, E. fetida Waki may have commercial application for biomass utilization and as a dietary health supplement.