Characteristics of wine produced by mushroom fermentation.

Saccharomyces cerevisiae is the main microorganism used in wine brewing, because this microbe has potent ability to produce alcohol dehydrogenase. We have recently discovered that some genera of mushroom produced alcohol dehydrogenase, and made wine by using a mushroom in place of S. cerevisiae. The highest alcohol concentration in this wine was achieved with Pleurotus ostreatus (2.6 M, 12.2%). In the case of Agaricus blazei, the same alcohol concentration (1.7 M, 8%) was produced under both aerobic and anaerobic conditions. This wine produced by A. blazei contained about 0.68% beta-D-glucan, which is known to have a preventive effects against cancer. The wine made by using Flammulina velutipes showed thrombosis-preventing activity, giving a prolonged thrombin clotting time 2.2-fold that of the control. Thus, the wine made by using mushroom seems to be a functional food which can be expected to have preventive effects against cancer and thrombosis.

Characteristics of a cheese-like food produced by fermentation of the mushroom Schizophyllum commune.

Lactate bacteria of the Lactobacillus and Streptococcus genera are normally employed in cheese making because these microbes have potent ability to produce lactate dehydrogenase. A milk-clotting enzyme is also necessary to make cheese. Recently, we discovered that some mushroom genera produce both lactate dehydrogenase and a milk-clotting enzyme. Using the mushroom Schizophyllum commune in place of a lactate bacterium, we produced a cheese-like food that contained about 0.58% beta-D-glucan, which has been shown to have preventive effects against cancer. The food also exhibited thrombosis prevention activity, prolonging the thrombin clotting time to 49.6-fold that of the control.

Development of mushrooms for thrombosis prevention by protoplast fusion.

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 satisfactory. We therefore used protoplast fusion between L. sulphureus and the commonly cultivated basidiomycete Hypsizygus marmoreaus to obtain cultivable basidiomycetes that produced an anti-thrombin substance. For the protoplast fusion of L. sulphureus and H. marmoreaus, the protoplast concentration, alternating electric field intensity, dielectrophoresis duration, and field pulse intensity used were of 1 x 10(7) protoplasts/ml, 100 V/cm.1 MHz, 60 s, and 8 kV/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.

Structure of the human lactate dehydrogenase B gene.

Human genomic clones containing parts of the lactate dehydrogenase B (LDH-B) gene (approx. 25 kb in length) were isolated and characterized. The protein-coding sequence of human LDH-B gene is interrupted by six introns at codons nos. 42-43, 82, 140, 198, 237 and 278-279, and the positions of these introns are homologous to those of LDH-A genes from man and mouse. The 5' non-coding region of human LDH-B gene is interrupted by an intron six nucleotide residues upstream of the ATG translation-initiation site, whereas those of human and mouse LDH-A genes are interrupted at 24 nucleotide residues 5' to the ATG initiation codon. As is the case of LDH-A genes from man and mouse, there is no intron in the 3' non-coding region of human LDH-B gene.