Development of fermented black soybeans with thrombosis inhibitory activity using mushroom mycelia

In recent years, thrombosis, such as myocardial infarction and cerebral thrombus, has become more prevalent and is now recognized as a public health issue. Black soybeans fermented using Schizophyllum commune and Hericium erinaceum exerted strong fibrinolytic effects, while those using Laetiporus sulphureus and S. commune exhibited antithrombin activities. Black soybeans fermented using S. commune, H. erinaceum, and L. sulphureus exhibited antioxidant activities that are 3-, 2.3-, and 2-fold stronger than that of a control （black soybeans）. Therefore, black soybeans fermented using S. commune possess the capacity to inhibit thrombosis through fibrinolytic, antithrombin, and antioxidant effects. Protease was involved in S. commune fermentation, proteins in black soybeans decomposed to some extent and contained many amino acids and peptides as well as essential amino acids. Furthermore, allergen protein levels were reduced, while those of nucleic acids, a component of umami, were increased.departmental bulletin pape

Study of alcohol dehydrogenase involved in alcohol fermentation by Basidiomycetes.

Production of alcoholic beverages using basidiomycetes has been reported. Alcoholic beverages produced by these basidiomycetes have also been reported to have various functionalities possessed by basidiomycetes. The capability of basidiomycetes to produce alcoholic beverages through fermentation may provide advantages. Alcoholic beverages with functionality derived from basidiomycetes can be given added value. However, the basidiomycetes used for the production of alcoholic beverages have no common feature of family and genus. The superior basidiomycete for producing alcoholic beverages is unknown. In this study, a wide range of basidiomycetes with ADH suitable for alcoholic fermentation were screened using various mushroom mycelia. The results indicate that basidiomycetes have both ADH involved in ethanol synthesis and decomposition, and that mushrooms with only ADH involved in ethanol production may be suitable for alcoholic beverage production. L. nuda had a high specific activity of ADH involved in ethanol synthesis, and Native-PAGE showed only the active band of ADH involved in ethanol synthesis. From the perspective of ethanol production, ADH that consume ethanol are to be avoided, thus mushrooms with ADH primarily involved in ethanol synthesis are suitable targets for alcoholic beverage production.departmental bulletin pape

Ergothionein Production by using Bonito Soup with S.commune and Suppression of Browning.

担子菌の菌糸体中にはエルゴチオネインが存在していることが報告されているが、菌糸体中のエルゴチオネインが培養液中へ分泌される可能性については報告されていない。そこで、本論文ではスエヒロタケの菌糸体中でのエルゴチオネイン生産量や培養液へのエルゴチオネイン分泌について検証した。10週間の培養終了後、菌糸体中および培養液中から得られたエルゴチオネイン量はそれぞれ8.2mgと1.1mgであり、菌糸体中のエルゴチオネイン量は培養液中のエルゴチオネイン量よりも7.5倍多かった。すりおろしたリンゴに添加した場合においても褐変防止効果がみられた。さらに、エルゴチオネインの標品よりも、スエヒロタケ菌糸体の無細胞抽出液として用いた方が酸化防止効果が高いことも明らかにした。鰹出汁培地で培養したスエヒロタケ菌糸体の無細胞抽出液が酸化防止剤として利用できる可能性を見出した。It has been reported that ergothionein is present in mycelia of basidiomycete cultured in liquid.However, there is no report on the possibility that ergothionein in the mycelium is secreted into the culture medium after culturing. Therefore, in this study we examined ergothionein production in Schizophyllum commune mycelium and ergothionein secretion into the culture medium. After cultivation, the production of ergothioneine obtained from the mycelium and culture broth was 8.2mg and 1.1mg, respectively, and the production of ergothioneine in the mycelium was 7.5. It was twice as much. The cell-free extract containing ergothionein prepared from the mycelium obtained was extremely high in antioxidant activity, and even when it was added to the apples, browning prevention effect was observed. Furthermore, it was also revealed that the antioxidant effect was higher when using it as a cell-free extract of S.commune mycelia than the standard product ergothionein. We have found the possibility that cell-free extract of S.commune mycelium cultured in bonito soup broth can be used as antioxidant.departmental bulletin pape

担子菌の発酵能による機能性大豆食品の開発

We evaluated the physiological activity of soybean fermented with mushroom mycelia．Without heating，fibrinolytic activity was observed in Schizophyllum commune（1418mm2），Pleurotus cornucopiae（ 1635mm2），Hericium ramosum（1608mm2）and NAPA（Ganodermataceae produced in Thailand） （1284mm2）． The cell-free crude extract form Schizophyllum commune（563mm2）and Pleurotus cornucopiae（ 369mm2）showed the fibrinolytic activity．Concerning the antithrombin activity，the thrombin clotting time was more than 600 seconds for Coriolus versicolor，Pleurotus cornucopiae，Lenzites betulina，Wolfiporia cocos，Schizophyllum commune， NAPA， Hericium erinaceum，Wynnea gigantea， Ramaria botrytis，Cordyceps militaris，Hericium ramosum and Stropharia rugosoannulata. Even after heating，Wolfiporia cocos and Macrolepiota procera showed the clotting time of more than 600 seconds. Compared with the blank（soybeans before fermentation），the total amino acid level increased to 38-fold for Schizophyllum commune and Pleurotus cornucopiae，to 20-fold for Hericium ramosum and to 15-fold for NAPA. In addition，the possible conversion from the glycoside type of isoflavone to the aglycon type was suggested

Development of functional Okara-Miso by mushroom fermentation

豆腐製造時に生じるおからは、その多くが産業廃棄物として処理されており、有効利用が望まれている。 おからを原料として味噌製造を試みることとしたが一般的な味噌作りで用いられているコウジカビではおからの繊維質を分解することが困難であることが、過去の研究で明らかにされている。そこで、セルラーゼやプロテアーゼ、さまざまな機能性を持つ担子菌を用いて、機能性おから味噌の製造を試みた。 ムキタケ（W-２）はプロテアーゼ活性だけでなく、コウジカビには認められなかった高いセルラーゼ活性を有し、特に50％おからを配合した試料において高い活性が認められた。さらに50％おからを配合し食塩を発酵３週目に加えることで、遊離アミノ酸や抗酸化活性が増加し、より旨味のある機能性味噌の製造が可能であることが明らかとなった。またセルラーゼ活性が高値を示し、味噌中の残渣が減少したことから、ムキタケを用いることでコウジカビでは困難であった機能性おから入り味噌の製造が可能であることを確認した。Okara is generated during tofu production. Most of okara have been treated as industrial waste, so effective utilization of okara is desired. However, it has been demonstrated in previous studies that it is difficult for Aspergillus oryzae（NBRC5238）which are used in a general miso making to decompose the fibrous okara. Therefore, we tried the preparation of miso containing okara having functionality using basidiomycetes which was produced protease, cellulase and variety of functions. Sarcomyxa serotina （W-２） has not only a protease activity, high cellulase activity, which was not observed in A.oryzae. Especially, higher activity was observed in the sample prepared with blending 50% okara. Then, the addition of salt to the samples prepared with 50% okara after ３ weeks cultured was fermented by S.serotina, free amino acids and antioxidant activity is increased, it became clear that it is possible to produce a functional miso a more taste. The cellulase activity indicates a high score and the residue in miso was reduced. So, it was confirmed that it is possible to manufacture a miso containing functional okara, which is difficult by using the A.oryzae, by using the S.serotina.departmental bulletin pape

魔法の酵素 ―きのこ酒誕生? そして新たなる挑戦―

departmental bulletin pape

The ClpXP ATP-Dependent Protease Regulates Flagellum Synthesis in Salmonella enterica Serovar Typhimurium

The ClpXP protease is a member of the ATP-dependent protease family and plays a dynamic role in the control of availability of regulatory proteins and the breakdown of abnormal and misfolded proteins. The proteolytic activity is rendered by the ClpP component, while the substrate specificity is determined by the ClpX component that has ATPase activity. We describe here a new role of the ClpXP protease in Salmonella enterica serovar Typhimurium in which ClpXP is involved in the regulation of flagellum synthesis. Cells deleted for ClpXP show “hyperflagellate phenotype,” exhibit overproduction of the flagellar protein, and show a fourfold increase in the rate of transcription of the fliC encoding flagellar filament. The assay for promoter activity of the genes responsible for expression of the fliC showed that the depletion of ClpXP results in dramatic enhancement of the expression of the fliA encoding sigma factor ς(28), leaving the expression level of the flhD master operon lying at the top of the transcription hierarchy of flagellar regulon almost normal. These results suggest that the ClpXP may be responsible for repressing the expression of flagellar regulon through the control of the FlhD/FlhC master regulators at the posttranscriptional and/or posttranslational levels. Proteome analysis of proteins secreted from the mutant cells deficient for flhDC and clpXP genes demonstrated that the ΔflhD mutation abolished the enhanced effect by ΔclpXP mutation on the production of flagellar proteins, suggesting that the ClpXP possibly defines a regulatory pathway affecting the expression of flagellar regulon that is dependent on FlhD/FlhC master regulators

The Recent Development of the Surgical Treatment for Hepatocellular Carcinoma

The optimal treatment for hepatocellular carcinoma (HCC) should be selected based on tumor conditions, liver functional reserve, and performance status. Surgical treatment, such as liver resection and liver transplantation, is the most favorable treatment method; however, its indication criteria differ according to each country’s guidelines. In Western countries, liver resection is indicated only for early-stage HCC patients with Barcelona-Clinic Liver Cancer staging classification (BCLC) 0/A. While in Asian countries, liver resection is one of the treatment options for advanced HCC, such as BCLC B/C. Recently, the treatment of HCC is about to enter a drastic transitional period. It started with the widespread use of minimally invasive surgery for HCC, followed by a high rate of hepatitis C virus eradication with the advent of direct acting antivirals and developing a multidisciplinary treatment for highly advanced HCC. As a result, the importance of liver resection for HCC is increasing, and it is time to reconsider the criteria for selecting treatment methods for HCC patients. This article outlines current topics in the surgical treatment of HCC

A novel prediction model of pancreatic fistula after pancreaticoduodenectomy using only preoperative markers

Abstract Background Since clinically relevant postoperative pancreatic fistula (CR-POPF) can cause intra-abdominal hemorrhage and abscesses, leading to surgery-related deaths after pancreaticoduodenectomy (PD), its preoperative prediction is important to develop strategies for surgical procedures and perioperative management. This study aimed to establish a novel prediction model for CR-POPF using preoperative markers. Methods On a training set of 180 patients who underwent PD at the Yamaguchi University Hospital, a combination of CR-POPF predictors were explored using the leave-one-out method with a unique discrete Bayes classifier. This predictive model was confirmed using a validation set of 366 patients who underwent PD at the Osaka University Hospital. Results In the training set, CR-POPF occurred in 60 (33%)　of 180 patients and 130 (36%)　of 366 patients in the validation set using selected markers. In patients with pancreatic ductal adenocarcinoma (PDAC), the main pancreatic duct (MPD) index showed the highest prognostic performance and could differentiate CR-POPF with 87% sensitivity and 81% specificity among 84 patients in the training set. In the validation set, the sensitivity and specificity of the MPD index-based model for 130 PDAC samples were 93% and 87%, respectively. In patients with non-PDAC, the MPD index/body mass index (BMI) combination showed the highest prognostic performance and could differentiate CR-POPF with 84% sensitivity and 57% specificity among 96 patients in the training set. In the validation set, the sensitivity and specificity of the MPD index/BMI-based model for 236 non-PDAC samples were 85% and 53%, respectively. Conclusion We developed a novel prediction model for pancreatic fistulas after PD using only preoperative markers. The MPD index and MPD index/BMI combination will be useful for CR-POPF assessment in PDAC and non-PDAC samples, respectively