α-Amylase from a still culture filtrate of Tricholoma matsutake, an ectomycorrhizal fungus, was isolated and characterized. The enzyme was purified to a homogeneous preparation with Toyopearl-DEAE, gel filtration, and Mono Q column chromatography. The α-amylase was highly purified (3580 fold) with a recovery of 10.5% and showed a single protein band by SDS-PAGE. The enzyme was most active at pH 5.0-6.0 toward soluble starch and stable within the broad pH range 4.0-10.0. This α-amylase was a relatively thermostable enzyme (optimum temperature, 60°C; thermal stability, 50°C). The molecular mass was 34 kDa by size-exclusion chromatography and 46 kDa by SDS-PAGE. This enzyme was not inhibited by the Hg 2ϩ ion. Measurement of viscosity and TLC and HPLC analysis of the hydrolysates obtained from amylose showed that the amylase from T. matsutake is an endo-type (α-amylase). Substrate specificity was tested using amylose with different polysaccharides. This α-amylase readily hydrolyzed the α-1,4 glucoside bond in soluble starch and amylose A (MW, 2900), but did not hydrolyze the α-1,6 bond and cyclic polysaccharides such as α-and -cyclodextrin.

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Transplantation of matsutake-infected pine tree into a pine forest has great potential as an artificial cultivation method of Tricholoma matsutake. Matsutake-infected pine trees had previously transplanted by this research group into the experimental area from 2001 to 2004 and its survival rate determined in 2006 was 20%. For the survived samples, the characteristics of mycelial growth and the development of fairy-ring formation of T. matsutake have been evaluated until 2009. It was found that size of the indeterminate ring showed significant differences among the individual trees and varied from 4 cm x 4 cm to 52 cm x 35 cm. The variation was possibly resulted from the differences in production area of the matsutake-infected pine tree and those in site characteristics of the transplanted spots. For the characteristics of mycelial growth, it grew in the shape of a cudgel or an acuteangled 'V' in early stage, and then the shape became more wider and changed into an obtuse-angled 'V' as time passed. We expect that matsutake mushrooms may occur from the fairy-rings of some of these matsutake-infected pine trees in the autumn of 2010.

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Omphalotus japonicus is known to as a poisonous mushroom in East Asia, but Illudin S, bioluminescent compound, has been recently reported for an anticancer effect. The fungus is quite rare in natural condition in Korea. Therefore, we tried to cultivate in the mushroom on the sawdust cultivation to provide sufficient amount of mushroom for pharmaceutical study for the future. The mushroom could be harvested from all sawdust media used in this study, and the sawdust of Pinus densiflora was considered to be the best for mushroom cultivation with considering mushroom productivity and cultivation period. The mushroom was produced 43~80 g fresh weight per 600 g sawdust medium in average, and could be harvested one to three times during cultivation period.