山梨大学醗酵研究所 研究報告[1254年-1997年]
JOURNAL OF THE INSTITUTE OF
ENOLOGY AND VITICULTURE

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vol.11 (1976)
「ブドウ酒際酵中の酵母について (第9報) 選択培地による野生酵母の分離 (その3)」
Dynamic Aspect of Yeast-flora during Vinous Fermentation
Part 9 Preferential Isolation of Wild Yeasts(Ⅲ)  
小原巌,野々村英夫,山崎豊彦 
Yuwao OHARA, Hideo NONOMURA and Toyohiko YAMAZAKI
pp.1-11[PDF]
  1)前報に引続きSALおよびMAA両培地を野生酵母の選択培地として使用し, ブドウ酒果酸の醗酵中における野生静母群の動態を知るため醗酵初軌中期および後期別に分擬された酵母群別にその分布状態 (生菌数の割合)を求めた。
 2)分離した547株の代表菌株34妹のうち29珠は7属11株と1変種に同定されたが,4株はS.florentinsの類縁菌とし,1株は同定されるものがなくCandlda spp.とした。それらのうちにはブドウ果醪から分康された報告のないC.guilliernonilii var. membranaefaciens(4株) と稀にしか分離されない Schizosaccaromyces pombe(2株), および別に新種として発表したTorulopsis vinacea(4株)があるほかには特に珍らしいものはない。
 3)両培地で測定した野生群母の生菌数は初め10⁵/ml程度であったが,醗酵が進むに従って次第に減少する。特に S0₂を多く (150ppm)加えた場合はその傾向が著るしい。
 4)これらの野生酵母を群別にみると C.krusei,K.apiculata,T bacillarisは 醗酵初期には多いが中期までにほとんど消減したのに対しT.vinacea とS.roseiは終期まで多数に残存する。 しかしS0₂の多い場合S.roseiは中期までに減少し, それに代ってSc.ludwigiiが多くなった。
 5)生菌数は徴畳ではあるが醗酵終期まで Sz.pombeおよびC.kruseiが分離されたほかT.vinaceaは初期から終期まで最も優勢な野生辞母の一つであることを明らかにした。
 The frequency of occurrence of wild yeast flora during fermentation of white must containing 50ppnL (No.1) and 150PPuL (No2) of sulfur dioxide were studied with a synthetic agar containing l-1ysineasso-esource of nitrogen and a malt agar containing 20ppm of actidion. The representative 20 pure cultures out of 524- strains isolated atrandom, and the 14 cultures of 23 strains obtained collectively were identified according to the system of LODDER and KREGR -VAN RU and classified in 14 groups (species) as follows: Rhodotorula mucilaginosa(A),Candida spp.(B),C.krusei(C).C.guilliermondii var. membranifaciens(D),C. albicans(E),C. tropicalis(F), Torulopsis vinacea(G),T.bacillaris(H),Kloeckera apiculata(I),K. africana(J),Schizosaccharomyces pombe(K),Saccgarimycodes ludwigii(L),Saccharomyces rosei(M), S.florentinus(N).
 It was shown that the initial populations of these yeasts were about 1.5×10⁵ per ml of must No.1 and 0,5 x 10⁵ per ml of No.2, and decreasing in numbers toward later. During the initial stages of fermentation, C, G, H, I and M predominated in must No,1 whereas in No,2 only G and M were isolated prevalently. On the other hand, L developed during the intermediate phase while M disappeared (in must No.2), however G servied, and a minute number of C and K was found until later. In this respect, it was reported that the occurrence of K was worth recognizing and also G was the one most prevalently isolated.
「リンゴの発酵的利用に関する研究 (第8報)リンゴを原料とする甘味果実酒の試醸」
Studies on Fermentative Processing of Apple Fruit
Part 8. Experimental Production of an Apple Wine or Sweet Cider from Apples.
増田博,四条徳崇,村木弘行
Hiroshi MASUDA, Noritaka SHIJO and Hiroyuki MURAKI
pp.13-17[PDF]
 リンゴを原料とする甘味果実酒として,アルコール16°,糖濃度10% apple wineおよびアルコール10°,糖濃度40%のSweet ciderを,それぞれ発酵軌上法および発酵完結後の混成法の2種の方法で試解した。その結果apple wineの方が良いきき酒結果を示すはかりでなく,貯蔵管理も容易でSweetciderよりすぐれていること, また謹塑造方法に関しては,必ずしも本格ポートのような発辞停止法にこだわる必要はなく,通常の果実酒にアルコール,白糖,果汁等を添加して混成する方法を用いても,少なくとも新酒については大差のない酒質が得られることを認めた。また各試良酒について,その分析値を示した。
 Two types of sweet beverages were experimentally produced from Kogyoku variety apple , one (A) was an apple wine which contained approximately 16 per cent of alcohol and. 10 per cent of sugars, and the other (B) was a sweet cider which contained approximately 10 per cent of alcohol and 4 per cent of sugars. For each type, two methods of producticn. were carried out, i. e. arresting the fermentation by fortification (A-1) or by sulfiting followed by cooling (B-1), and sweetening by cane sugar (A-2) or by apple juice (B-2) after completion of the fermentation From the results of analyses and organoleptic taste-testing, these sweet beverages were found to have superior qualities. The apple wines were rather preferred to sweet ciders. These methods of production, however, seemed to have no serious effects on the organoleptic qualities and natural composition of the ciders obtained.
「リンゴの発酵的利用に関する研究 (第9報)トマトピュレーを添加したリンゴ酒の試醸」
Studies on Fermentative Processing of Apple Fruit
Part 9. Experiments With the Addition of Tomato Puree the Apple Juice in Cider-Making
増田博,四条徳崇,村木弘行
Ooki NAKAYAMA and Hiroko KOIKE
pp.19-26[PDF]
 リンゴ酒の香味改良の試みとして, トマトの混和離進を行なった。その結果,混和量が適当であれば,異香味を与えることもなく,コク味を増してリンゴ酒の酒質改良に有効であることを認めた。ただし過度に多く混和すると濃色と異香味を生じて結果がよくない。 またトマト混和により,多少清澄しにくくなり,滓の量もふえるが,10%程度の添加量では障害となる程ではない。 トマトを添加するとリンゴ果汁の発聾は促進され,残糖分は減少する.エキス分,全窒素,遊離アミノ酸窒素, ペプチド鮎窒素はいずれも増加する。P.C.によりアミノ酸組成を検討した結果,通常のリンゴ酒からはアスパラギン酸, アスパラギン, リジン, アルギニン, グルタミン酸,セリン, グリシン, スレオニン, グルタミン,α-アラニン,チロシン, プロリン,バリン, メチオニン, ロイシン (イソロイシン)および未確認のスポット1個が見出され,トマトを添加すると, グルタミン酸,グリシン,スレオニン,α-アラニン,チロシン, ロイシン (イソロイシン)の量が増大する他, フェニルアラニン,β-アラニンおよび未確認のスポット1個が新たに生ずることが認められた。 また P.C.による有機腰の検索結果, クエン酸, リンゴ酸, コハク酸,乳酸を確認した他,グリコ-ル酸に近いRfを与える酸が見出され, トマト添加によってクエン酸およびグリコール酸相当の酸が増加することを認めた。たゞし酒のpHはほとんど変化しない。  
 A tomato puree(Bx.115) was added to apple juice before fermentation in the ratio of 10 and 25 percent by volume, and the qualities of the resulting ciders (C-10,C-25) were compared with the normal cider (C-0).  
 The fermentation of the apple juice became rapid and vigorous. As the result, the ciders gave reduced sugar contents. While the amounts of total nitrogen,free amino-nitrogen and peptide-nitrogen in the ciders were increased considerably. By paper chromatography, fifteen amino acids (i.e aspartic acid, asparagine, lysine, arginine, glutamic acid, serine, glycine, threonine, glutamine, α-alanine, tyrosine, proline, valine, methionine and leucine) and one unidentified spot were detected in the normal cider(C-0). Whereas it was observed that in the cider(C-25) two amino acids (i.s.phenylalanine and β-alanine) and one more unidentified spot further appeared, besides the amounts of glutamic acid, glycine, threonine,α-alanine, tyrosine and Leucines were increased.  
 As for the organic acids in the ciders, citric, malic, lactic, succinic and glycolic (?) acid were identified. Among these acids, citric and glycolic (?) acid were found to be more abundant in the cider(C-25) than in the normal cider (C-0).  
 The results of the organoleptic taste-testing showed that the addition of 10 percent tomato purge gave a superior dry cider with rich body and no off odors, while the addition of 25 percent unfavorable.
「リンゴの発酵的利用に関する研究 (第10報) リンゴ酒の加熱処理について」
Studies on Fermentative Processing of Apple Fruit
Part 10. Thermal Treatment of Ciders
増田博、四条徳崇、 村木弘行
Hiroshi MASUDA, Noritaka SHIJO and Hiroyuki MURAKI
Pp27-35[PDF]
 通常のリンゴ酒,リンゴ・ブドウ混醸酒,トマトピュレーを添加して窒素成分を増したリンゴ酒,およびショ糖を添加したリンゴ酒について,それぞれ60℃で60日間加熱処理を行い,分析値の変化および利き酒結果を示した。リンゴ酒については、加熱処理によってベーキング・シェリー酒のような特異の芳香の生成を期待することはできず,またコク味の不足を補なうこともできないが,焦臭に似た1種の芳香を附与することはできる。処理リンゴ酒は,単独では必ずしもすぐれた酒質のものとはいえないが,他の酒類に対する混和剤としての利用価値は認めてよいと思われ,たとえばベーキング・シェリー酒に対する少量の混和は,それ程の酒質の劣化を生じない。しかも原料としてブドウよりは安価である。
 加熱処理によって,リンゴ酒中の揮発酸,揮発エステル, アルデヒド等が増加し,不揮発酸タンニンおよび色素,窒素成分の減少がおこるが,これらはブドウ酒の場合と全く同様であった。
 Eleven ciders prepared experimentally in the previous papers, i. e. a normal cider, a series of ciders blended with up to 75 per cent by volume of grape wine, two ciders enriched in nitrogenous compounds by the addition of tomato purée before fermentation, and a cider sweetened by sucrose, were baked for 60 days at 60°C, and the results of the chemical analyses and the organoleptic taste-testing of the treated ciders were shown in comparison with the corresponding untreated controls.
 Baking sherry-like beverage such as from grape wine could not be obtained from any ciders, but a burnt smell could be somewhat given to the baked ciders, especially in the presence of sucrose. These baked ciders were rather of blending use with other wines or spirits.
 It was just alike in the case of grape wines that the appropriate amounts of volatile acids, volatile esters and aldehydes in these ciders were increased, while that of fixed acids, tannins, coloring matters and nitrogenous compounds were decreased by the treatment.
「酵母菌態を利用するブドウ酒の醸造について」
Utilization of Yeast Cells in Wine-Making
 Part 2. Mechanical Disintegration of Yeast Cells in Wine Lees with Decompression Rupture for improving the Quality of Table Wines
増田伯、村木博之
pp. 37-43[PDF]
 甲州種ブドウ果およびブドウ酒母OC-2を用いて醸造した白ブドウ酒の滓を、ドライアイスを用いて凍結し、過剰のドライアイスと共にステインレス製耐圧タンクに入れ、密閉して温度を上げ、融解させるとともに炭酸ガスによって61-63kg/㎡の圧力を生じさせこれをノズルから急激に噴出せしめた後、遠心分離によって上澄みを採取してブドウ酒長に液を製造した。この処理によって滓中の酵母菌体が酵素による分解作用を受けやすくなるため、酵素添加によって窒素成分の溶出を増加させると共に、処理液からの上澄収量を増大させることができる。しかし調味液としての酒質改良効果は、単に勤怠破壊処理を加えたのみのものの方がやや優れていた。
  この方法は処理に長時間を要せず、また滓の酸化、汚染、酵母臭の発生等の防止が容易で、優れた滓の利用方法と思われる。
「赤ブドウ酒の色調に及ぼすペクチナーゼの効果について」  
The Effect of Pectinase on the Depth of Color of Red Wine
曽根公平、加賀美元男、大村智、高山正則
pp. 45-51[PDF]
  Black Queen種のブドウを原料として赤ブドウ酒を醸造する際,ペクチン分解酵素 (スクラーゼ)を応用し,その添加量および作用時間等を異にする7区分の試醸酒について, 主として色調の変化を比較検討した。
 スクラーゼの添加により色調は濃くなるが,その効果は必ずしも添加量に比例しなかった。 しかし工業的に使用できる0,02%程度の添加量でも一応の効果が認められ, また色素の抽出はスクラーゼの最適温度(45℃)で作用させ1-2日間皮発酵させたのち添加する方法が効果的であることを明らかにした。 メタノール生成量はスクラーゼ添加で僅かに多くなる有毒飲食物等帯締令による許容量(1mg/ml)には達しない程度であった。
「ブドウ果汁の処理工程における粘性挙動について」
Effects of Certain Physical and Chemical Conditions on the Viscosity of Grape Juice
鈴木康夫、加賀美元男
Yasuo SUZUKI and Moto-o KAGAMI
pp. 53-58[PDF]
 ブドウ果汁の流動性に関する基礎的資料を得るため、二、三の処理工程における年生挙動について比較検討した。
 1)果汁の殺菌などの目的により加熱された場合,その前後において粘度に及ぼす影響を調べた結果,ほとんど影響のないことが認められた。
 2)果汁の示すpH変化に伴う粘性への影響は, pHの上昇に従って粘度も増加する傾向をを示した。
 3)果汁中の主成分である糖濃度の変化と粘度との関係を追究すべく,加熱濃縮および稀釈による変化の影響を調べた結果、本実験における糖濃度の範囲において粘度は糖濃度の指数函数で表わされることを明らかにした。
 4)ペクチン含有量の相異による粘度変化を検討した結果,ペクチンの増加に伴い粘度は増加し,その傾向はペクチンの多いほど大であることが認められた。
 5)温度差による粘度変化は糖濃度の大なるものほど,その温度依存性の大なることが明らかにされた。
 6)ブドウ糖, ショ糖、ペクチンおよび酒石酸の4物質からなる混合水溶液を調製し,果汁類似液としてその粘度を測定し果汁のそれを比較した結果,果汁の粘度はこれら4種の物質によってほとんど決定されるものと思われる。
Under certain physical and chemical conditions comparable to practical cellar operations, various factors affecting the variation in viscosity of grape juice were studied.The results obtained are as follows :
 (1) The heat treatment of the grape juice have little effect on the viscosity.
 (2)The Viscosity Of Juice Increased with pH range from 2 to 4.2 and 、with the pectine contents.
 (3) A linear relationship between the Viscosity and the Sugar concentration in grape juice was obtained on semilogarithmic coordinate. 
(4) The susceptibility of viscosity to temperature of juice obviously appeared in the case of high sugar concentration.
 (5) According to the relationship between the viscosity of grape juice and of the modal juice, it seems likely that the variation in viscosity is practically indeed by glucose, suclose, pectine and tartaric acid contains.
「低酒精飲料の研究 (第1報)弱補糖果汁から出発した低酒精ブドウ酒の試醸」
Studies on the Low Alcoholic Beverages
Part 1. Experimental Production of Soft Table Wines from slightly Sugared Grape Musts
櫛田忠街,中山大樹,小池弘子
Tadae KUSHIDA, Ooki NAKAYAMA and Hiroko OKOIKE 
pp.59-64[PDF]
糖分20%まで補臆した1963年度の MuscatBaileyA(MA)種および甲州種のブドウ果汁を用いて,低酒糖分甘味ブドウ酒の試軸をおこなった。残糖が約7%になったとき,冷却(6-7℃1l週間)遠沈および火入れ (65℃,1分間)によって発酵を抑制し, 防腐剤としてサリチル/酸 (0.2g/L),パラオキシ安息香酸ブチル,(0.25g/L),メタカリ (0.20g/L),デヒドロ酢軟ソーダ (0.05g/L)またはソルビン酸 (0.05g/L)を加え,2L瓶詰めて約1年間貯蔵し,化学分析および男女別の利き酒をおこなった。その結果,次の成績を得た.
1)発酵停止処理をしないものおよび冷却処理し,防腐剤を加えないものは,糖分が喰い切られた。
2)冷却処理したものは,1週間の冷却期間中に 2 (甲州種)または 3 (MA種)%程度、糖分を消費したが,加熱区は,処理した時のままの状態を一年間保った。
3)防腐剤は,いずれも有効であったが,中でメタカリは製品を脱色する鯛向があり,またソルビン酸は褐変を促進する傾向がある。
4)防腐剤を添加しないものは一般に香味が劣り, また甲州瞳の場合、冷却処理したものにくらべて,加熱処理したものの方が,香味が安定していた。
 Two procedures of arresting the fermentation when the must has fermented to the point (7 per cent residual sugars) were compared, that of centrifuging the chilled (6-7℃) musts, and that of pasteurizing for 10 minutes at 60℃. After storage for a year, effect of various antiseptics on the chemical and organoleptic quallty of soft table wines obtained were examined.
 Residual Sugars of the pasteurized wines were not decreased, but of the centrifuged wines, especially of the wlnes without antiseptics were somewhat decreased. Al l antiseptics used, i.e.salicylic acid, paraoxybutylbenzoate, potassium metabisulfite, Sodium dehydroacetate and sodium sorbate, were satisfactory in preventing undesirable challges during the storage, except sorbate which accelerated browning of the pasteurized wines, and potassium metabisulfite which somewhat decolorized the red wines.
「ブドウ酒醸造における酸化防止剤の利用に関する研究
(第2報) 白ブドウ酒に対するアスコルビン酸類と亜硫酸との混用試験」
Studies on the Use of the Antioxidant in Wine Making
柳田忠衛,丸山智章,吉田紘一
Tadae KUSHIDA, Chiaki MARUYAMA and Koichi YOSHIDA
pp. 65-69[PDF]
  常法の如く醸造した甲州産白ブドウ酒を使用してアスコルビン酸(C)またはエリソルビン酸(E)S0を各種の割合に混用した場合のブドウ酒品質に与える影響を比較検討した。主なる結果は次の通り。
 1) 前報でも認められたように,CまたはE加の多いほど,S0₂残量が多い。しかし,S0₂の添加量とCの残量との間には一定の関係がない。つまり,S02がCの酸化分解を防止するのか,または促進するのか,今回の実験では不明であった。
 2) 吸光度よりみると,S0₂と混用したCまたはEの効果は,D430においてはS0₂の効果にかくれてしまってほとんど認められないが、D530においては大きくあらわれた。結局,全色度はS0₂ Cまたは Eとの混合割合によって変化するが、それらの値より概算すると,200ppmの Cまたは Eは大体 100ppmのS0₂の脱色効果に匹敵するようである。
 3) 利き酒の結果ではS0₂Cまたは Eを混用したものはいずれも大差がなかった.従って,混合割合はどの位がよいか、にわかには決定できないが、SO₂無添加のもの,C単独使用のものは成績が悪いので,必要最低限のSO₂使用量があるように思われる。
 Effects of I-ascorbic (C), erithorbic (E) and sulfurous acid (SO₂) on the absorbance, chemical composition and olganoleptic quality of white wines prepared from Koshu variety were examined.
 An appreciate amount of SO₂ in the wine were found in accordance with the increased amound of C (A) added during storage, however, there was little correlation between the content of C (or E) and the amount of SO₂ added. A distinct effects of C(or E) on the decrease in absorbance at 530nm (D530) were showed, but that on the D430 were not observed owing to the presence of SO₂. The sum of the absorbance at 370-610 nm of the wine varied, accordingly, with both amount of C (or E) and SO₂ added.
 The olganoleptic quality of the wines added with a varied amount of two. antioxidants, e.g.C and SO₂, was superior to that of the wines added with C (or E) alone or with them.
「梅および梅酒中のシアン化水素について」
Studies on the Hydrogen Cyanide inthe Ume (Japanese plum) and Ume-shu (Japanese plum liqueur)
田中健太郎,飯沼静子
Kentaro TANAKA and Shizuko IINUMA 
pp. 71-76[PDF]
 数回の仕込み結果より考察すると、仕込みの際35°アルコールに砂糖を完全に溶解したのち、原料と混和すること、この方法により、果実成分の抽出が速やかに行われ、梅酒中の青酸含量は約15日~20日で最高となるが、約1か月後には少なくなり、味、香りともに良いものが得られる。
 青酸中毒については、致死量が60mgと言われているが、これは青酸含量の最も多い20日前後のものでも約9Lに相当することとなり、飲料として適当となる仕込後1か月以上のものについては、中毒は全く考慮する必要はないと思われる。
 利き酒結果より見ると、完熟期の梅を原料としたのもが、味、香り共に最もよく、未熟期のものからは種子成分のうち、ベンズアルデヒド臭が特に乏しく、またアルコール臭も残り、苦みも強く不適当である。このことは種子中の成分が梅酒独特の香味を作り出す上に大きな影響を与えているものと考えられる。
 次に梅酒中の青酸含量が仕込後20日以降は徐々に減少するが、生産が如何なる状態で消失するかについて、先づ賛成の梅酒中から徐々に揮散することが考えられたので、これをアンモニア水に吸収させる装置を作って検討したら、現在のところ確認されていない。また他の有機物と結合して、梅酒中に他のかたちで存在することも充分考えられるので、今後検討を行うつもりである。
pp.77-82[PDF]
ブドウ酒のマロラチック醗酵に関する研究 (第3報) リンゴ酸分解最近の分離及び果醪への試用(英文)
Studies on Malo-lactic Fermentation
III. Isolation of Malo-lactic Organisms and Inocula-~ion of the Isolates into Musts
野々村英夫、小原巌、加賀美久、風間敬 Hideo NONOMURA, Yuwao OHARA, Hisashi KAGAMI and Kei-ichi KAZAMA:
Nippon Jozo Kyokai Zasshi (J. Soc. Brew. Japan) 59, 513 (1964)
 Three hundred and twenty four strains of malo-lactic organisms were isolated from 26 samples of musts and lees (representing 7 grape varieties) with ROGOSAS tomato juice-liver extract medium modified by the addition of 20ppm actidione, 20ppm eurocidine,500ppm sorbic acid and 2% agar. The ratio of the organisms to total isolates increased in the progress of must fermentation. And it was usually greater in red wine lees than in white examined. The representatives, 45 strains, of the isolates were classified into 4 groups (included 9 isolates) of Lactobacillus, and 6 (36 isolates) of Leuconostoc. Nine strains representing 8 groups out of above 10 were inoculated into the red musts (Black Queen and Merlot) and the white musts (Koshu) respectively, each must was 20l in volume, and contained 50 ppm S0₂, 23 or 25 % sugars and 2 % starter (ca.10⁶cells/ml). A strain of heterofermentative Lactobacillus and a strain of Leuconostoc (sp. related to citrovorum) were effectively induce the fermentation in all t!1e three kinds of musts, and the latter strain seemed to cause the fermentation with the greatest regularity. The wines from the musts, in which the fermentation took place by these strains, were lower in acidity, higher in pH value, and better in flavor and taste than the control wines.
日本産ブドウ酒酵母に関する研究(第8報)
山梨県産Flor Yeastsの分類(英文)
Studies on Japanese Wine Yeasts
VIII. Classification of Flor Yeasts Isolated in Yamanashi Prefecture, Japan.
横塚勇、両角劦喜
Isami YOKOTSUKA and Shuki MOROZUMI Zasshi (J. Soc. Brew. Japan) 57, 836 (1962)
Nippon Jozo Kyokai
 Thirty three typical strains of f10r yeasts were selected from 847 cultures isolated from 470 samples collected at C6 wineries and several vineyards in grape and wine producing areas.
They were classified in accordance with the procedures and systems of LODDER and K. -van RIJ as follows: 5 strains of Saccharomyces cerevisiae; 6 of S.rosei ; 2 of S.steineri ; 2 of S.exiguus; 3 of S.helerogeniclis and 15 of S.ovijormis.
  Following two noticeable characteris were observed for all strains tested: (1) fseudoffijceliums were not formed and
(2)Protuberances resembling to conjugation tubes were always present on the cultures on GORODKOWA agar slant or gypsum block.
日本産ブドウ酒酵母に関する研究(第9報) Flor Yeastsの発酵期における生理学的諸性質(英文)
Studies on Japanese Wine Yeasts
IX. Physiological Properties in Fermentation Stage of Flor Yeasts Isolated in K6fll Valley, Japan
横塚勇
Isami YOKOTSUKA Ibid., 57, 943 (962)
  The comparison of properties in fermentation stage between flor yeasts (I) and non-film forming yeasts (II) was conducted by using 33 strains of 1 and 51 of ordinary II isolated from many wines made by spontaneous fermentation.
 The data obtained confirmed the results of many workers, and indicated that 1 resembled ordinary II in their temp. for growth or fermetnation, their tolerance to S0₂ or pH values, their fermentation velocity, and their abilities of producing alcohol, organic acids, esters and aldehydes. But the results of testing of their wines showed that bad yeasts seemed to be more abundant in 1 than in II and that good ones. were more in II than in I.
日本産ブドウ酒酵母に関する研究(第10報)
Flor Yeasts Sherry醸造並びにDry Table Wineの品質改善(その1)
Flor YeastsのFlor Filmと被膜細胞について(英文).
Studies on Japanese Wine Yeasts
X. Utilization of Flor Yeasts to Flor Sherry Making and Improvement of the Quality of Dry Table Wine. (1) Flor Film and Its Cells of Flor Yeasts.
横塚勇 Isami YOKOTSUKA: Ibid., 57, 1197 (l962)
 Twenty nine strains isolated in Kofu valley, three (Jerez-5, Xeres-l and Xeres-2) of typical Spanish and two (D-Douroveu and E-Evora) of Portuguese for yeasts were cultured on dry wine, and film formation, type of film and its cells were observed as follows;
 (1) Each strain had a special type of film which was classified into five groups.
 (2) Excessive film formation occured frequently on wine contg below 14% EtOH under favorable environmental condition, which deteriorated the quality of wine.
 (3) Cells of film did not essentially differ from that in the fermentation stage. They gathered into a flock, but did not form a pseudomycelium.
「日本産ブドウ酒酵母に関する研究(第11報)
Flor Yeasts Sherry醸造並びにDry Table Wineの品質改善(その2)
皮膜形成化のブドウ酒の成分並びに香味の変化(英文)」
Studies on Japanese Wine Yeasts
XI. (2) Changes Brought about in the Composition and the Flavor of Wine by the Flor.
横塚勇、後藤昭二
 Twenty nine strains of Japanese and five of foreign yeasts mentioned above were cultured on dry dessert wine and dry table wine, and the changes brought about in the compn. and the quality of wines by the flor were estd. as follows: There was no much noticeable difference in the changes brought about in the compn. of wines by the f1or of between the Spanish yeasts and the others. Flor character developed noticeably and the aldehyde (I) increased to between 150 and 250mg/l by the fourth to fifth week. The strength of flor character increased as the amount of total 1 increased. But it is likely not the amount of total 1 but the amount of fixed 1 that has strong effect. The amount of EtOH used by the f10r was only below 1%.
 In most of the cultures volatile acids decreased to below 0.2g/l one year after inoculation. A fall in fixed acidity ranged between 0.5 and 2g/l. The pH values of the cultures did not change so much. The amount of sugar used by the flor seemed to be very little. EtOH, glycerol, acetic acid or lactic acid were likely a good carbon source for film growth. Total esters increased a little, but the amount of neutral esters did not almost change. Total I increased remarkably and in some case it amounted to about 800mg/l after one year. Fixed 1 increased in wine as total I increased, but not in a constant ratio.
「日本産ブドウ酒酵母に関する研究(第12報) Flor Yeasts Sherry醸造並びにDry Table Wineの品質改善(その3)Flor Yeasts の被膜形成に及ぼす外囲の培養条件とブドウ酒組成の影響:培養温度、ブドウ酒の酒精含量とpH値の影響について(英文)」
Studies on Japanese Wine Yeasts
XII. (3) The Influence of Environmental Factors and the Composition of the Wine on the Growth of Flor Film: Culture Temperature, and Alcohol Content and pH Value of the Wine.
横塚勇
Isami YOKOTSUKA and Shoji GOTO Ibid., 58, 169 (963)
 The influence of incubation temp., the ale. content and the pH value of the wine on film formation (1) of flor yeasts were examd. under lab. conditions using 27strains of Japanese and five of foreign yeasts montio:1ed above. The results obtained were as follows:
 (1) Temp.: On table winos conig. less than 13% EtOH, the rapid Ⅰ occured at temp. up to 28°C with most strains used. It has been proved by many investigators that the optimum temp. for Ⅰ on wines contg. between 14 and 15% EtOH is about 20°C, and that at temp. higher than about 23° to 25°C, Ⅰ does not often occur. At ION temp. below about 15%, Ⅰ was usually very slow. At about 10°C on wines contg. less than 12% EtOH, one third of the strains used failed to develop as a fi1m and at temp. down to between 5° and 7℃ all strains did not form complets films, and, Ⅰ, if it occurred, was restricted to scant, small patches.
 (2) The ale. strength of the wine: An optimum ale. content for Ⅰ by most strains used was between 13 and 14%, while the max, for complete films with some strains was above 15%. In this respect four strains resemble quite closely MARClLLA'S typical Spanish sherry yeasts. The rate of Ⅰ was increased by lowering the ale. content, but Ⅰ at such low strengths were too much, very unstable and apt to sink.
 (3) The pH value of the wine: The optimum pH values for Ⅰ was between 3.0 and 4.0. As the pH values of most Japanese ordinary wines are between 3.0 and 4.0, so it is unnecessary to adjust the pH value for Ⅰ.
pp. 83-87[PDF]
Studies on Japanese Wine Yeasts XIII. (4) The Quantities of Sulfur Dioxide and Sugar in Wine.
Isami YOKOTSUKA, Shoji GOTO and Yoshihide YAMAKAWA: Ibid., 58. 1221 (1963)
 The effects of the quantity of S0₂ added and sugar in wine on the film formation (I) by flor yeasts in Delaware white table wine contg. below 12% EtOH were examd. and the results obtained were as follows:
 (1) The toxity of S0₂ on I was very strong. It could not be checked by culture temp. or ale. concn. of wine.
 (2) The tolerance of I could not be raised by training.
 (3) The quantity of sugar used in I on table wine was so small, and usually below 0.2%.
 (4) Good I was observed on the wine, which contd. only below 0.1% of reducing sugar estd. by BERTRAND'S method and therefore was lacking in fermentable sugar. No active I was caused by adding sugar in the wine on which flor yeasts had completely developed, but it occurred when small quantity of new wine contg. almost no sugar. Accordingly there seems to be better growth factors than sugar.
Studies on Japanese Wine Yeasts XIV. (5) The Concentratio: J of Ethanol in Wine.
Isami YOKOTSUKA, Shoji GOTO and Yoshihide YAMAKAWA: Ibid, 59, 84 (1964)
 The effect of the strength of EtOH on the development of flor character and the changes in the component of wine brought about by flor was studied. The flavor was tasted and components were analysed on four kinds of dry white wine; contg. 15.0, 14.3, 13.5 and 12.5% EtOH, after being stored under the films of Jerez-5, WF-107, W-210 and W-500 for one year respectively. Results obtained were as follows:
 (1) The amount of EtOH consumed by the flor was SJ small that it was not a serious problem from economical point of view.
 (2) Both total acids and non-volatile acids were consumed the most by the f10r in the wines contg. 15.0% EtOH and the least in the wines contg. 12.5% EtOH. The amounts of volatile acid; consumed by the flor for all wines tested were so much that they amounted from 60 to 70% of their initial amounts. The strength of EtOH-I under vigorous flor seemed to be check the flor to consume acids in wine.
 (3) Both the accumulation velocity and the accumulated amount of aldeh, de (I) under vigorous flor were generally not affected by the strength of EtOH-I.
 (4) The flor character (flavor) of the wines under flor increased to some extent as the amount of 1 increased. It was found that the amount of 1 accumulated did not dominate the flavor, and that the flavor depending on the strains. The amounts of I, in which superior flavor bad always developed under the film of Jerez 5, were always less than those in the wines under the films of the others.
Studies on Japanese Wine Yeasts XV. (6) Culture Temperature.
Isami YOKOTSUKA, Shoji GOTO and Yoshihide YAMAKAWA Ibid., 59, 163 (1964)
 The effect of culture temp. on the quality of wines: under flor was investigated by observing the film formation, tasting flavor and analysing the components of white dry table wines contg.12.8% EtOH, on which the films of Jere-5 and WF-107 had covered for eight months at 10°, 15°, 20° and 25°C. Results obtained were as follows:
  (1) No film formation at 10°C. Slow and a little formation at 15°C, but complete film cover was never made through the whole period studied. Very rapid at both 20° and 25°C; complete film cover after 20days and thick films after 30days, and then the films grew too much.
 (2) There was no effect of temp. on the consumption of EtOH by the flor. The consumption of volatile acids increased as temp. rose, but no remarkable difference was recognized in the amount of total acids consumed. The accumulation, velosity and the accumulated amount of aldehyde increased as the temp. rose.
  (3) After one month flor character was developed in all wines except the wine incubated at 10°C. The flavors of the wines incubated at 20° and 25°Cwere equally strong, but that of the wine at 15°C was comparatively weak.
 After 40 days they were all satisfactorilly strong, but after 50days the flavor of the wines incubated at 25°C, smelled of yeast and was inferior to the other wines. It is worth to notice that sufficiently strong flor character was developed in the wines on which the film were only 10 to 15%. The complete thin film cover seemed to be best.
Studies on Japanese Plum Liqueur
 Ⅰ.The influence of Manufacturing Processes upon the Quality and the Composion of Organic Acids d Liqueur.
Isami YOKOTSUKA and Shoji GOTO: Nippn Jozo Kyckai Zasshi (J. Soc. Brew. Japan) 59, 633 (1964)
Six kinds of the liqueurs(I)were made by the following different processes:
 (a) Moderately ripe plums and sucrose were put in approx,35 % EtOH for about two months, then the extract was separated, bottled and matured at room temp.
 (b) After plums were put in 35% EtOH for two months, the ext. was drawn off, then sucrose put on the same plums for' the same period, and the ext. sepd. Both ext. were blended, and the mixture was bottled and matured. 
 (c) Completely crushed plums were used.
 (d) Extn. was done with approx. 50% EtOH.
 (e) Over-ripe plums were used. (f)Extn. was done for one year. The compn. of I produced was examined by a general chern. analysis, organic acids (II) in I were detd. by column chromatography, and I tasted. Thus the influence of the pricesses, applied upon the compn. esp. of II, and the quality of I were studied. The results obtained were as follows:
 (1) There was only little difference in the yields of I.
 (2) Considerable changes were observed in the amounts of II, except malic acid, present in I. Their amounts depended to some extent on the processes, but the same kind of II were always detected in all I. The content of malic acid was considerably affected by the processes.
 (3) The kinds and the amounts of II detected in I by column chromatography were approx. as follows (expressed in % to total acids): citric acid 55, malic acid 33 as a mean value, unidentified acid 10, oxalic acid 2, and acetic acid 1.5.
 (4) The quality of I examd. by tasting was as follows: In 10 months after bottling I (a) was ordinary. I (b) was the best. I (c) was somewhat too sour and rough in taste and smelled of seeds. In 20 months after bottling: I (c) had improved considerably in quality. I developed strong fragrant flavour, and was rich in bod; and rounded in taste. I (b) was almost as good as I (c), but it was somewhat flat and watery in taste. I (e) developed strong fragrant flavour, but was fairly cloud; in appearance. Both I (d) and I (f) had always fairly cloudy appearance and bad off-odors.
Studies on Japanese Wine Yeasts XVI. (7) Strains of Flor Yeasts.
Isami YOKOTSUKA, Shoji GOTO and Yoshihide YAMAKAWA Ibid., 59, 267 (1964)
 The effects of 27 strains of Japanese and three of Spanish yeast mentioned before, on the quality of both dessert wine and table wine under their films for 90 days were investigated by tasting the wines and analysing their components, and the conclusions were obtained as follows:
 (1) The Strains of the yeasts, whose oxidative activities and decompg. abilities on acids at their film stage were too strong or too weak, could not develop strong fragrant flor character, and most strains of Japanese yeasts tested fell into this category in their abilities.
 (2) Only the strains, which oxidized alc. to aldehyde, and aldehyde to acids to adequate degrees, and then decompd acid, moderately, much improved the quality of the wine3 by reducing the acidity and developing strong fragrant flavor.
Studies on Japanese Plum Liqueur  I. The influence of Manufacturing Processes upon the Quality and the Composition of Organic Acids Liqueur.
 Isami YOKOTSUKA and Shoji GOTO:Nippn Jozo Kyckai Zasshi (J. Soc. Brew. Japan) 59, 633 (1964)
 Six kinds of the liqueurs(I)were made by the following different processes:
 (a) Moderately ripe plums and sucrose were put in approx,35% EtOH for about two months, then the extract was separated, bottled and matured at room temp.
 (b) After plums were put in 35% EtOH for two months, the ext. was drawn off, then sucrose put on the same plums for' the same period, and the ext. sepd. Both ext. were blended, and the mixture was bottled and matured. 
 (c) Completely crushed plums were used.
 (d) Extn. was done with approx. 50% EtOH.
 (e) Over-ripe plums were used.
 (f)Extn. was done for one year. The compn. of I produced was examined by a general chern. analysis, organic acids (II) in I were deted. by column chromatography, and I tasted. Thus the influence of the pricesses, applied upon the compn. esp. of II, and the quality of I were studied. The results obtained were as follows:
 (1) There was only little difference in the yields of I.
 (2) Considerable changes were observed in the amounts of II, except malic acid, present in I. Their amounts depended to some extent on the processes, but the same kind of II were always detected in all I. The content of malic acid was considerably affected by the processes.
 (3) The kinds and the amounts of II detected in I by column chromatography were approx. as follows (expressed in % to total acids): citric acid 55, malic acid 33 as a mean value, unidentified acid 10, oxalic acid 2, and acetic acid 1.5.
 (4) The quality of I examd. by tasting was as follows: In 10 months after bottling I (a) was ordinary. I (b) was the best. I (c) was somewhat too sour and rough in taste and smelled of seeds. In 20 months after bottling: I (c) had improved considerably in quality. I developed strong fragrant flavour, and was rich in bod; and rounded in taste. I (b) was almost as good as I (c), but it was somewhat flat and watery in taste. I (e) developed strong fragrant flavour, but was fairly cloud; in appearance. Both I (d) and I (f) had always fairly cloudy appearance and bad off-odors.
Studies on Japanese Plum Liqueur  II. Identification of Two Unlmcwn Organic Acids found In Japanese Plum Liqueur.
Isami YOKOTSUKA and Shoji GOT0 Ibid., 59, 636 (1964)
 Two unknown organic acid; (I) found in the liquer were studied by paper chromatography, column chromatography, gas chromatography, and infrared spectrum analysis. The identification of the neuteal compounds fron I, and the comparison between the amount of alkali used to neutralize definite amounts of I and that used to saponify them were also made from results obtained in these expts., these two I were presumed to be monoethylcitrate (II) and its isomer, respectively.
 Next, II was synthesized and its pure crystals were obtained and demonstrated. Then the same expts. as mentioned above were, repeated using pure crystals of synthetic II, and the data obtained in the former expts. and in the latter ones were compared. Thus it was concluded that I were II and its isomer, respectively. II was found for the first time in the liqueur in this study.
Studies on the Application of Ion Exchange Process to Fermentation Industries XIII. On the Stabilization~ of Metallic Deterioration of Wines (Part 1)
Moto-o KAGAMI : J-Iakko Kogaku Zasshi (J. Ferlli. Tech. Japan), 42, 565 (1964)
 The present paper describes the initial steps in an investigation of the effectiveness of the chelating resin on table wines contg. 20mg/l Cu and 10mg/l Fe. The behavior of some typical ions in the wines were investigated using three forms (H, Na, Mg) of the resin (Dowex A-1) with batch process. It was shown that the adsorption attains equilibrium slowly, and of the cations, Cu was removed to the largest degree followed by Fe, Ca and Mg, however K was removed scarcely. It has been found that the treatment is not only detrimental to the flavor but in many case the wines appear to have improved in organoleptic characters.
Studies on the Application of Ion Exchange Process to Fermentation Industries XIV. On the Stabilization of Metallic Deterioration of Wines (Part 2).
Moto-o KAGAMI and Satoshi OMURA: Ibid., 42. 571 (1964)
The effect. of the chelating ion exchanger in the three forms mentioned above on the removal of Cu added to table wines was examined. Treatmeat of wine with the resin was carried out by passing at the rate of 10-30 v.v.h. through a columns contg. 7.5ml and 40ml of the resin. It was found that treatment in all three forms removed Cu very effectively from the tested wines e,pe:ially from white wine.
Studies on Fermentative Processing of Apple Fruit I. Experimental Production of a Fruit Wine from Apples Combined with Grapes.
Hiroshi MASUDA, Noritaka SHTJO and Hiroyuki MURAKI: J-Iakko Kogaku Zasshi (J. Ferill. Tech Japan), 41, 655 (1963)
 In this study a fruit wine was exptl. produced from a combination of apples (I) and grapes (II). The results are summarized as follows:
  (1) High acidity of II in Japan was lowered by the addn. of I. When malo-lactic fermentation occurs in the wine, the acidity will be further lowered by the consumption of malic acid which is replaced with II tartaric acid by the addn. of I. Thus the most adequate acidity can be expected in the resulted wine.
  (2) Nitrogenous compd. and tanninsin the wine were inevitablly of less quantities, than pure grape wine.
  (3) The mixing of both fresh juice; followed by fermentation gave better quality to the resulting wine than the mixing of both after the fermentation. By the former method. a ratio of less than 50% I juice gave good results in 0rgganoleptic taste-testing.
  (4) The max. absorptions at about 230 and 320mμ of grape wine diminished by the addn, of I. This diminution was consistently related to the quantity of I added in the case of fermentation after mixing, and irregularly in the case of mixing after sep. fermentation.
Studies on Fermentative Processing of Apple Fruit
II. On the Maceration of Fresh Pulp in Cider-Making.
Hiroshi MASUDA, Noritaka SHIJO and Hiroyuki MURAKI Ibid., 42, 7 (1964)
 In the process of cider-making, the effects of the maceration (I) of the crushed apple pulp for 20hrs. were tested in two ways ; one was in an oxidative state by contact with air and the other was in a nonoxidative state using S0₂ , The yield of juice was increased by I in both ways, The oxidative I decreased acid and tannin contents in the juice by the action of the enzymes in apple tissues, Probably due to the high acidity of the Kogyoku variety, the activity of polyphenol oxidase and the decrease of tannins was not so remarkable and caused no difficulty in normal cidermaking. The decrease of acids reached about -45% compared with the unmacerated juice, and the resulting cidor abo had a reduced acid content. Sugars in the juice also decreased but volatile acids increased slightly, The non-oxidative I caused no decrease in the acid content, but conside rable increase in the tannin content in the juice and the resulting cider. The scores of organoleptic taste-testing showed the good effects of I on the quality of cider, especially in the case of the oxidative I. From these results it is concluded that I of the fresh pulp may be recommended for the apple varieties with relatively high making. The decrease of acids reached acidity.
Studies on Fermentative Processing of Apple Fruit
III. On Defecating (Keeving) Cider.
Hiroshi MASUDA, Noritaka SHUO and Hiroyuki MURAKI:Ibid., 42. 11 (1964)
 The most suitable method for defecating cider from the apples with a considerably high acidity. was investigated. It was performed by the addn. of CaCO₃ and pectic enzymes "Sclase" to the juice and the cooling of the juice to 5-7°C. The farmentation of the brown head and the clarification of the juice was complete, and the obtained cider was naturally sweet and gave the best results in organoleptic taste-testing. Both CaCO₃ and pectic enzymes were indispensable for successful I defecation. The use of CaHPO, 2H₂O instead of CaCO₂ was undesirable. The yield of cider was lowered through complete defecation.
Studies on Fermentative Processing of Apple Fruit
 IV. Fermentation of Milled Apple Pulp in Cider-Making.
Hiroshi MASUDA, Noritaka SHIJO and Hiroyuki MURAKI: Ibid. 42, 379 (1964)
 Different ciders were exptl. produced by the fermentation of crushed apple pulp. Thick wort of just crushed apple pulp with difficult to ferment, and addn. of apple pomace (I) to the pressed juice in a ratio below 50% is recommended. The ciders obtained by this method had rich body and intense aroma of apple fruit. Addn. of 35 to 50% I gave the best score, in organoleptic taste-testing. Nitrogenous compd. and tannins in the cider increased and fixed acid decreased by the addn. of I. The decrease of fixed acids was greater with the addn. of smaller (5% for example) amt. of I. These changes produced no undesirable effects on the quality of the ciders.
Studies on Fermentative Processing of Apple Fruit
 V. Use of Complex Yeast Cultures in Cider-Making.
Hircshi MASUDA, Noritaka SHIJO and Hiroyuki MURAKI :Ibid, 42, 383 (964)
 Various kinds of apple juice were fermented with Saccharomyces cerevisiae (I), S.rosei (II).Kloeckera Kloeckera(III) and Torulopsis bacillaris (IV) separately and in various combinations. Analyses of the resulting ciders showed that the yield of EtOH was highest with I alone and considerably low with III and IV; II produced the most aldehyde and volatile esters; IV formed a remarkable amt. of volatile acids; III caused a considerable decrease in the fixed acid content. With the mixed cultures of I: III and I: IV in proportion of 2: 1, it was observed that III or IV depressed the fermenting activity of I. and the yield of EtOH was lowered while considerable amt. of residual sugars remained in the ciders. In organoleptic taste-testing, the use of IV and I alone and in combination with each other gave the best scores. By these results it was shown that natural sweet cider of good quality may be obtained by the suitable use of the mixed yeast cultures.
Studies on Fermentative Processing of Apple Fruit
VI. Production of Brandies from Apple Pomace.
Shoro AMEMIYA, Shuki MOROZUMI, Noritaka SHIJO and Hiroyuki MURAKI: Ibid., 42, 388 (1964)
Apple brandies were exptl. produced from the apple pomace (I). A brandy made from the macerates of I had similar flavor to that from the normal juice, while the fermentation of the thick wort of I gave a brandy with inter.se aloma of apple fruit. The quality of these brandies was quite high and showed that I may be used as a component of superior brandies. The yields and analytical data of the apple juice. I macerates, material ciders, and brandies were shown.
Studies on Fermentative Processing of Apple Fruit
VII. Experimental Production of a Sherry-type Wine from Apples.
Hiroshi MASUDA, oritaka SHI]O and Hiroyuki MURAKI : Ibid., 42, 548 (1964)
 An apple juice mentioned above was coned. to a sugar content of 25 %, racked from the sediment, and fermented with a sherry yeast "Jerez-5". Three years later the resulting wines were blended and aged in three kegs after the model of the "solera system". During the storage, the yeast formed only thin and small islands of film on the wines, but the finished wine had the characteristic flavor somewhat of "flor". The addn. of film growth factors has a possibility to improve the quality of the wine. Fixed acids in the wines decreased during the storage, while aldehydes, volatile acids and esters increased. The oxidative maceration of the crushed pulp may be recommended to reduce the acidity of the juice so as not to be too high after the concn. A good apple wine similar to a brown sherry can be obtained by the above process.
酵母による鉄含有物質の生合成に関する研究 (第1報)Saccharomyces cerevisiaeにおける鉄の消長について 酵母による鉄含有物質の生合成に関する研究
Studies on Biosynthesis of Iron Containing Substances by Yeasts.
 Part 1. The Fate of Iron in Saccharomyces cerevisiae
四条徳則、田中健太郎、川西康博、伴野文男:
 日本農芸関東支部大会講演(1964年4月1日 於東京大学農学部)
Noritaka SHIJO, Kentaro TANAKA, Yasuhiro KAWANISHI and Fumio TOMONO
 ⁵⁹Fe(I)をトレーサーとして、過量のFe⁺3をブドウ果汁に加え、ブドウ酒発酵を行ったところ、酵母が鉄を菌体内に取り込み。更にこれをブドウ酒中に鉄含有物質として排出することが推測された。そこで人工培地にⅠ-Fe⁺3を加え、これにS.cerevisiae (OC-2) を加え、振盪培養し、経時的に資料を取り、菌体と培養液に分離したノリ、それぞれ放射能(Ⅱ)、菌体数、菌体重量およびpHを測定した結果、培養の初期には、菌体内のFe⁺3は徐々に増加するが、その後時間の経過とともに、培養液中の鉄含有量も増加してくる。約20時間で菌体内のⅡは最高となり、培養液中のⅡは約32時間で最高となり、以降はほとんど変化しない。次に前述の方法で培養し酵母のⅡ取込量が最高となる32時間で菌体を分離し、これをⅠ-Fe⁺3 を含まない培養液で同様に培養したところ、菌体内のⅡは、時間の経過と共に減少し、酵母と培養液中に排出された鉄含有物質について菓子部吸収スペクトル、ペーパークロマト等の実験を行ったところ、初めの培養液に加えたFe⁺3は、酵母により資化され、Fe⁺₂コンプレックスの形で排出されることが認められた。
山梨大学大学院 総合研究部附属 ワイン科学研究センター
The Institute of Enology and Viticulture, University of Yamanashi
所在地:〒400-0005 山梨県甲府市北新1丁目13-1

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