クリカエシ カイフンシキセイシュ ジョウゾウ 二 オケル コウボキンタイナイ エス アデノシルメチオニン

一段仕込みの醪及び固液共存培地の両方を用い,酵母の増殖プロセスと発酵プロセスを区別してSAM動向を検討した。酵母を過剰接種して増殖を廃すると,低温下における酵母菌体内SAM高蓄積などの含硫物の特徴的な動向の差は縮小傾向となった。これにより低温下での酵母菌体内へのSAMの高蓄積には,低温で増殖することの影響が大きいことが明らかとなった。また,酵母の過剰添加は醪の発酵にも大きく影響し,特に醪初期のボーメの低下とアルコール生成が早期化し,結果的に発酵期間は数日短縮された。 また,この醪から酵母菌体を遠心分離により回収し,次醸造へ再利用する回分醸造を試みた。回分醸造でも増殖プロセスを廃して順次スケールダウンして進めると,3回目または4回目以降では発酵に支障はないものの,アミノ酸度と製成酒中のSアデノシルメチオニン(SAM)の増加が顕著であり,酒質も明らかに異なり,味のくどさや苦味が感じられた。その一方,回収菌体を繰り返し利用しても,酵母菌体内にSAMが高蓄積され続けることはなく,1回目の醸造以降に極端な増加が認められることはなかった。 さらに,固液共存培地を用いて比較したところ,糖などは徐々に供給され並行複発酵しているが,醪初期のように米粒溶解に起因する固形物の減少や流動性の向上などの変化がないためか,発酵経過とSAM蓄積は回分醪ほどの差にはならなかった。このことから,醪初期の溶解に伴う流動性の向上及びボーメ低下がもたらす影響が重要であることが明らかとなった。The growth of yeast cells and the fermentation process were studied for the concentration of SAM in sake mash using sake mash and a model media, where the inoculum size was increased to suppress the growth. The result showed that the intracellular SAM concentration was highly accumulated at low tem-perature in sake mash and the model medium. Harvested yeast cells were collected by centrifuging of the mash and reused for the next mashing repeatedly. The fermentation was repeated safely for 3 or 4 times. The concentrations of amino acids and SAM in produced sake increased, and the results of sensory tests of the produced sake showed that the quality of produced sake was more thick and bitter according to the number of times the yeast cells were reused. On the other hand, intracellular SAM did not accumulate highly when the yeast cells were reused. The concentration of alcohol and SAM was compared during the repeated fermentation using the model medium. As a result, the difference of intracellular SAM in the model medium was smaller than that in sake mash. It was suggested that this result was caused by the existence of solid materials and the dissolution particularly in the early mashing period. This model medium was considered to be eective for the analysis of parallel fermentation in the early sake mashing process

セイシュロウ ト コエキキョウゾンバイチ ニ オケル コウボキンタイナイ ノ エス アデノシルメチオニン ノ ドウコウ

清酒醪中の酵母の存在部位別の菌体内成分について調べるために,従来の固液共存培地の寒天部形状を改変した。酵母は,従来通り固体部を斜面状に成形した培地において固体部の端のフラスコ器底ガラスが露出している部分に直接沈殿していることが観察された。これに対し,平板状に成形した改変培地では,器底部に形成した固体培地の上へ直接沈殿し,差別化が可能であった。培養中の液部のメチオニン濃度は,10℃の平板状固体培地でも低濃度一定で推移した。さらに15℃でも同様に一定濃度で推移する傾向を示した。菌体内S-アデノシルメチオニン(SAM)は,低温でかつ酵母菌体が固体部上へ直接沈殿する発酵条件で特に高蓄積が認められた。 次に醪中の酵母菌体を米粒に付着して存在する酵母と液部に浮遊する酵母に分画して回収した。その菌体内SAM濃度は,固体部に付着している菌体で高蓄積され,モデルの結果と合致した。一般に低温醪では米粒の溶けが悪く,粕歩合も高くなり,固形物が多くなることとの関連が考えられた。以上のように,低温醪で認められる酵母菌体内へのSAM高蓄積については,これまで明らかにしてきた醪中のメチオニンの低濃度推移から示唆される選択的な取込に加え,固液界面に存在する酵母が増加することが,更なる一要因として明らかとなった。SAMは,醪中で酵母菌体から漏出して製成酒の酒質に影響を与えることから,今後この改変培地を活用し,SAMの漏出と酒質について詳細な検討をすることが可能と思われる。The accumulation of SAM in yeast cells was examined. A modified model was developed to compare the yeast cells attached on the solid of the medium which was assumed cells in the rice fraction in sake mash. As reported previously, the yeast cells precipitated to the glass on the bottom of the ask in the medium. On the contrary, in this study the yeast precipitated on the surface of the at solid medium in the bottom of the flask. The yeast cells on the at solid medium accumulated high content of SAM at 10℃, whereas the methionine concentration in liquid medium was kept at low concentration. Furthermore, this characteristic tendency was also shown at 15℃. Next, SAM was highly accumulated in the yeast cells at the solid rice fraction of sake mash:this tendency was the same as that of the model medium. These results suggests that the high accumulation of intracellular SAM in the yeast cells was characterized as caused by the sake mash being at low temperature

セイシュロウ ジョウソウジョウケン ガ セイセイシュチュウ ノ エス アデノシルメチオニン ガンリョウ 二 オヨボス エイキョウ

S-アデノシルメチオニン(SAM)は,貯酒中にメチルチオアデノシン(MTA)に変換されて苦味を生じるが,特に醪末期でのアルコール添加と上槽中の経時変化について検討された例はない。そこで上槽モデルと酒造場からの試料採取によりSAMの酒質への影響について検討した。 アルコール添加によってアルコール濃度が上昇しても,末期品温管理が適切であれば,醪液部のSAM濃度が急上昇することは認められず,酒質に大きな影響はないと判断した。 製成酒中のSAM濃度は上槽中,経時的に上昇し,上槽圧力の上昇と同期していた。また酒造場垂れ口から採取した試料でも,酒袋を使用する酒槽式上槽機及び自動圧搾上槽機のいずれでも,経時的な上槽圧力の上昇に伴い製成酒中のSAM濃度は上昇した。 一方,酵母菌体を遠心除去した後に再混合して,酵母を含まない醪を調製し,これに圧力を掛けて上槽しても製成酒中のSAM濃度が上昇することから,上槽圧力の上昇が菌体からのSAM漏出に直接的に関与している可能性は低いと考えられた。しかし,粕部の遊離SAM含量は液部に比べて高いことから,圧搾によってまず固液分離中の醪から液部のみが徐々に減少し,固形物が相対的に増加した状態で強く圧搾されて酒粕を生じるという,物理的変化が要因として大きいものと推察した. さらに市販酒にSAMを添加して官能評価を行うと,原酒中に含まれる程度の濃度でも苦味として感じられることから,醪末期管理の重要性が示された。The influence of highly accumulated intracellular S-adenosyl-methionine (SAM) of yeast was examined on the quality of sake. Produced SAM was converted to methylthioadenosine (MTA) and had the effect of increasing the bitterness of sake. Then, the change of the concentration of SAM in the filtration process was examined. Especially, the alcohol addition to sake mash was studied with model filter press of the laboratory scale and compared with that of industrial scale of some sake breweries. The result showed the SAM content of sake was not affected immediately after alcohol addition. Therefore, it was suggested that the quality of sake was not influenced by temperature control of the late mashing process. On the other hand, it was clear that the concentration of SAM in produced sake increased during filtration according to the increase of the filtration pressure. The yeast cells were removed by centrifuging from sake mash, and the precipitates were mixed again without the yeast cells. After the filtration of this mixture, the concentration of SAM in the filtrate increased. This result suggests that the increase of SAM in produced sake was not caused by the leakage of SAM from yeast cells. Furthermore, the concentration of SAM in produced sake of sake brewery increased according to the pressure rise during filtration. This phenomenon was confirmed by the tests of 2 types of filter press. As the concentration of SAM in sake cake is higher than that of mash filtrate, the pressure of filtration is considered less likely to be involved in the leakage of SAM from yeast cells directly. In another examination, SAM was added to commercial sake and a sensory evaluation was carried out. The result showed that the increase of SAM in the sake was recognized to increase bitterness. It was suggested that the control of SAM of the late mashing process was important, and SAM content aected the quality of produced sake

タネコウジ セイゾウ オヨビ セイキク ニ オケル コウジキン Aspergillus oryzae ノ セイギョインシ KpeA ノ キノウ カイセキ

麹菌の転写因子KpeAは平板培養でのコウジ酸生産の制御因子として見出されたが，その後，広く二次代謝と分生子形成の制御に関わることが明らかとなった。種麹製造及び製麹は分生子形成の制御を伴う醸造工程であり，また，米麹中の二次代謝物は醸造物の品質や安全性に影響する。そこで本研究では，実際の醸造における培養工程である種麹製造と製麹において，KpeAの遺伝子破壊株と高発現株の形質を調べることでKpeAの醸造における役割を明らかにすることを目的とした。製造した種麹の状貌と遺伝子解析から，KpeAは，分生子形成の主要因子であるBrlAの遺伝子発現制御を介して，分生子形成を促進する役割を果たしていることが示された。製麹においても米麹に着生する分生子に差が生じたことから，KpeAは醸造工程においても分生子形成に重要な転写因子であることが分かった。一方，米麹中のコウジ酸量が破壊株で顕著に増加した。コウジ酸の推定生合成酵素KojAの遺伝子発現解析から，KpeAは平板培養と同様に，製麹においてもコウジ酸生産を転写レベルで制御することが確認された。さらに破壊株の麹抽出液は，コウジ酸が多く含まれるために褐変しにくいことが明らかとなったことから，KpeAの遺伝子破壊によるコウジ酸生産の増加が，米麹の褐変抑制に効果があることが示された。以上よりKpeAは種麹製造と製麹において，分生子形成やコウジ酸生産を制御しており，実際の醸造においても重要な役割を果たす転写因子であることが確認された。KpeAの遺伝子破壊株は，米麹の酵素活性が若干低下するものの，製麹において分生子が形成しにくく，かつ米麹の褐変性が低いといった清酒醸造に使用する麹菌として好ましい特性をもつため，KpeAは醸造における麹菌の育種ターゲットの新しい候補として期待できる。We previously identified a novel transcription factor, KpeA, in Aspergillus oryzae. KpeA is involved in secondary metabolism and conidiation when A. oryzae is cultured in liquid or agar medium. To clarify the role of KpeA in sake brewing, we analyzed the functionality of KpeA in rice koji making and seed koji making. In seed koji making, the kpeA disruption (ΔkpeA) strain showed longer aerial hyphae and the number of conidia was one-third that of the control strain. In accordance with conidiation, expression of brlA, a pivotal gene for conidiation, was also decreased in the ΔkpeA strain, whereas it was increased in the kpeA overexpression (OE) strain. These results suggested that, similar to the phenomenon that occurs in agar culture, KpeA induced conidiation via positive regulation of brlA expression in seed koji making.Rice koji samples made with ΔkpeA strain, OE strain, and control strain were analyzed for conidiation, enzyme activity, and secondary metabolites. The OE strain and the control strain formed conidia after 46 and 72 h of inoculation, respectively, whereas the ΔkpeA strain did not. In contrast, activities of representative hydrolytic enzymes in rice koji did not differ among the strains. Regarding secondary metabolism, we confirmed that kpeA is responsible for the enhanced kojic acid production via inducing the expression of the putative biosynthesis gene, kojA. Notably, the browning of rice koji extract was not observed in the ΔkpeA strain sample. We confirmed that additional dosage of kojic acid at the level of the rice koji of the ΔkpeA strain prevented the browning of rice koji extract of the control sample. As a result, kpeA disruption led to increased kojic acid amount in rice koji, which consequently prevented the browning of rice koji.Taken together, our results showed that KpeA regulates secondary metabolism and conidiation of A. oryzae during rice koji making and seed koji making. Although enzyme activities of the ΔkpeA strain were reduced slightly, we consider that the observed phenotypes, such as suppressed conidiation or repression of the browning of rice koji, are beneficial for rice koji or rice koji making. Thus, we propose that kpeA could be a new target for the breeding of A. oryzae for traditional brewing

ADAMTS-1: A metalloproteinase-disintegrin essential for normal growth, fertility, and organ morphology and function

金沢大学医薬保健研究域医学系A disintegrin and metalloproteinase (ADAM) represents a protein family possessing both metalloproteinase and disintegrin domains. ADAMTS-1, an ADAM family member cloned from cachexigenic colon adenocarcinoma, is unusual in that it contains thrombospondin type I motifs and anchors to the extracellular matrix. To elucidate the biological role of ADAMTS-1, we developed ADAMTS-1-null mice by gene targeting. Targeted disruption of the mouse ADAMTS-1 gene resulted in growth retardation with adipose tissue malformation. Impaired female fertilization accompanied by histological changes in the uterus and ovaries also resulted. Furthermore, ADAMTS-1(-/-) mice demonstrated enlarged renal calices with fibrotic changes from the ureteropelvic junction through the ureter, and abnormal adrenal medullary architecture without capillary formation. ADAMTS-1 thus appears necessary for normal growth, fertility, and organ morphology and function. Moreover, the resemblance of the renal phenotype to human ureteropelvic junction obstruction may provide a clue to the pathogenesis of this common congenital disease

Interferon regulatory factor-4 activates IL-2 and IL-4 promoters in cooperation with c-Rel.

Interferon regulatory factor (IRF)-4 is a member of the IRF transcription factor family, whose expression is primarily restricted to lymphoid and myeloid cells. In T-cells, IRF-4 expression is induced by T-cell receptor (TCR) cross-linking or treatment with phorbol-12-myristate-13-acetate (PMA)/Ionomycin, and IRF-4 is thought to be a critical factor for various functions of T-cells. To elucidate the IRF-4 functions in human adult T-cell leukemia virus type 1 (HTLV-1)-infected T-cells, which constitutively express IRF-4, we isolated IRF-4-binding proteins from T-cells, using a tandem affinity purification (TAP)-mass spectrometry strategy. Fourteen proteins were identified in the IRF-4-binding complex, including endogenous IRF-4 and the nuclear factor-kappaB (NF-κB) family member, c-Rel. The specific association of IRF-4 with c-Rel was confirmed by immunoprecipitation experiments, and IRF-4 was shown to enhance the c-Rel-dependent binding and activation of the interleukin-4 (IL-4) promoter region. We also demonstrated that IL-2 production was also enhanced by exogenously-expressed IRF-4 and c-Rel in the presence of P/I, in T-cells, and that the optimal IL-2 and IL-4 productions in vivo was IRF-4-dependent using IRF-4-/- mice. These data provide molecular evidence to support the clinical observation that elevated expression of c-Rel and IRF-4 is associated with the prognosis in adult T-cell leukemia/lymphoma (ATLL) patients, and present possible targets for future gene therapy

SRM・CRMの観点から考察する商社営業のコンピテンシー

Stem cells have captured the imagination of the general public by their potential as new therapeutic tools in the fight against degenerative diseases. This potential is based on their capability for self-renewal and at the same time for producing progenitor cells that will eventually provide the building blocks for tissue and organ regeneration. These processes are carefully orchestrated in the organism by means of a series of molecular cues. An emerging molecule which is responsible for some of these physiological responses is adrenomedullin, a 52-amino acid regulatory peptide which increases proliferation and regulates cell fate of stem cells of different origins. Adrenomedullin binds to specific membrane receptors in stem cells and induces several intracellular pathways such as those involving cAMP, Akt, or MAPK. Regulation of adrenomedullin levels may help in directing the growth and differentiation of stem cells for applications (e.g., cell therapy) both in vitro and in vivo. © 2012 Elsevier Inc.Peer Reviewe

Serial assessment of multimodality imaging in anti-leucine-rich glioma-inactivated 1 antibody encephalitis: A case report

In autoimmune encephalitis, abnormalities of diffusion-weighted imaging (DWI), fluid-attenuated inversion recovery (FLAIR), arterial spin labeling (ASL) in magnetic resonance imaging (MRI), single-photon emission computed tomography (SPECT) and 18F-fluorodeoxyglucose-positron emission tomography (18F-FDG-PET) have been reported. However, there are few studies of long-term follow-up of imaging. We report a case of anti-leucine-rich glioma-inactivated 1 antibody encephalitis whose MRI (DWI, FLAIR and ASL), 99mTcHM-PAO SPECT (PAO-SPECT) and 18F-FDG-PET were evaluated through the clinical course. ASL, PAO-SPECT and 18F-FDG-PET consistently showed abnormalities in almost the same area. Serial assessment of these imaging modalities is useful in evaluating disease activity and efficacy of treatment