Development of Mushrooms for Thrombosis Prevention by Protoplast Fusion

概要

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With thrombosis a major cause of death in Japan and the Western world, thrombin-inhibitory agents that constrain the formation of fibrin are sought. We screened for basidiomycetes showing anti-thrombin activity and isolated Laetiporus sulphureus. However, it was difficult to cultivate and its form was not satistactory. We therefore used protoplast fusion between L.sulphureus and the commonly cultivated basidiomycete Hypsizygus marmoreaus to obtain cultivate basidiomycetes that produced an anti-thrombin substance. For the protolplast fusion to L.sulphureus and H.marmoreaus, the protoplast concentration, alternating electric field intensity, dielectrophoresis duration, and field pulse intensity used were of 1×10^7 proplasts/ml, 100V/cm. 1MHz, 60s, and 8kV/cm, respectively. The number of regenerated colonies obtained was 4961, from which 43 strains were selected for electrophoretic analysis. Four of the fusants were found to have a band from each parent in isozyme patterns obtained using their crude extract. The fruiting bodies of the fusants were very similar to those of H.marmoreaus. Crude extract from each of the fusants and from L.sulphureus showed anti-coagulative activity in terms of the thrombin clotting time. We thus obtained improved basidiomycetes that produce an anti-thrombin substance, are easily cultivated, and whose form resembles H.marmoreaus, a commonly used culinary mushroom.
社団法人日本生物工学会の論文
2000-05-25