Fundamental studies on the utilization of olive fruits. III. The pickling of green olives (1) The biochemical changes during the fermentation of Spanish-type green olives-香川大学学術情報リポジトリ

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93. Voユuフ,No・・1(ユ9S5)

FUNDAMENTAL STUD‡ES ON THE UTILIZATION

OF OLIVE FRtTITS

IIIThe pickling of gIeen Olives(l)Thebiochemical

Changes duIing the fermentation of Spanish−tyPe green Olives

KenjiKArAElTRA and TeiichiNARASAKI

(Laboratory of AgriculturalProducts Technology) (Received MayごC,ユ955。Accep七edJunelO,1955。)

INTRO王)UCT‡0Ⅳ

01ives are utilizedin thIee Princiralways,Viz.,aSrlpe Olives,aS green Olives,and for olive oil TheTe are SeVeIalwaysbywhich greenolives mayl〕eutilized・・InJapan,theyareusedmainlya苧

、1\

ferrnented Spanish−tyPe greenOlives,and as showninthe first paper of this series,the yie王d of the green olives tends to develop year after year accompanied byincreasing biochemicalproblems

tol〕e SOlved,

〈2)

VAUG耶,et al. studied chemically and microbiologica11y the processesinvolvedin the production of Spanish・・tyPe gIeen Olives,eSpeCially七he fermentationprocess,and proposedvaluable recommen− datior)SJor tたein]PrOV(TrlentCfits productio‡1。They have divided the fermentation processinto

three stages,i.・e”:the pIimarystage of fermentation,Which persists noImally for7to14days,and containsthemicTOOrganismswhichTr)ay COntribute to deterioration of theolive$i董the fe工mentati−

On does rlOt PrOCeedin a no‡malmanner,nOtably gram−negativebacteria of the genera AeYOba ar!dPseud(nl{710Sandyeas†s;theinteTmediatestageoffeImentation,in which the predominating mic−

roorganisrr)S aIelactic acidbacteria,Leuconos10C and Lac!obacillus,the totalacidityincreasesrapidly, andthestagelasts for訟tOtltt\れO tOthIeeWeeks;andthe finalstageoffermentation,inⅥhichthe

totalacidityincreases from about〔).3 tol..C%aslactic acid,andthe high acidqtolerantlactob− acillipredomina.teT上eimFOrt8r)CeOf ttefirst stsgeof fermentation has been pointed out,and the use of the staIter and tte addit王on of supplenlentary EUgar tO brine solutions have also been discu− ssed.IzQt汀ERDO(S14)also dividedthe fermentationprocessinto three stages

Researches on the feTTr)er)tation processes of various−tyPe green Olives have been continued by COnSiderable workers.BofiBOLLA aIld his associates have studied the effects ofpH and acidity of

thebIine for the feIrnentation(5・6),t上euse of pure ct71tureof Z.aciobacillusinthe feImentation(7・S), and the effect of thelye tIeain7ent folloⅥed byvasIling([,10,‖).Chemicalcor)Stituentswereanaly2=ed in the residuallye solution(1ビ,1S),andan acidic white crystalline substance,C30H4SO3WaS Separ・a− ted(111〉l・A darkening of the brinesolutionof Spanish−tyPegreen Olives hadbeen studied(15),and a Substance,Which ぐauSed丑e darker2irg,ⅥaSミhoⅥn tO be precipitated by3C%1ead acetate.It was

recommended to n7aintain丑e acidity of thebriI二e SOlution at o5,−{9%aslactic acid to avoid the darkening.It has been alsoreported that the growth of naturalye島st flora on the sur・face ofbrine SOlution wasinhibitedby OいCく1L<.C∈%fenneloil(16),andthatfeェ1neloilcouldinhibitthe growthof

Aerobacieraerogenes,Which causes the gasFOCketsinSpanish−typegreen Olives(Ⅰ;〉InS。S.S.R,AFI(i8) has obtainedthe acidityof O.こ5%asiactic acidin amonthwiththeadditionofl%sugar and2% tomato juice,Ⅵhile the王1atUralbrineresulted onlyinthe acidityofO.1%aslactic acid even atthe end of three months of fermentation

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94 Te・CIl..Bull.Ⅸagawa Agr..Coll.

Thusthe differentphasesof thisindustry,Whichwerepracticedoriginallyinapurelyempirical

manneI,SeemtOhavetoday afirm scientific basisHoweverIthe results repoItedbyvarious aut− hors are rather considerably differentaccordingtothe authors,prIObablybecauset:hechemicalc〇m− positions of the o】ivesusedin each study vary accordingtothelocalitywheretheoliveshad grown

andthefactorsinfluencing the fermentationmay alsodiffer fromeachother.It seems,therefore, Withoutmeaningless toinvestigate theprdblemsinvolvedintheproduction of Spanish−type gr・een OlivesinJapan・Tofacilitユモe theproductioz10f Spユnishqtypegreenolives,itseems tobeneces$ary

toinvestigatethe chemicalandmicrobioiogicalchanges occurringintheprocessesoftheproduction,

especiallyin the fermentation process,and the pretr.eatments should alsobeinvestigatedin conn・

exion with the fermentation proces$

The authorswi11investigate the feごm3ntation ofSpまnish−typ3green Olives to establish the stand−

ardmethod of prepar’ationinthiscountry.,Biochemicalchangesoccurringinthenaturalbrinesolutions duringthefeImentationof Spanish−typegr・eenOlivesmayb∋describedinthepresentpaper

】姐A甘ERIAI.AND三城E甘盟0》

The MiSSion,Seuillano,and肋nzanillo varieties are a11useまfor・the produc七ionof Spanish−type green oiivesInthis country,the Missionvarietyisfavor・eiinits medium size and ease of prepaト

ation,and this varIiety wasusedin the present studies

Theolives weTe Picked aboutNovember2〕th,1954,in ShうdoIsland,COVeredwith dilute sodium

chloride solution,and sent to the authors′1aboratory for the experiments

Theolives wereplacedin shaliow vat;aてId coveredwith2%sodiumhydr・OX.ide solutionat roc)m temperature‥The$01utionwas aIlowedto penetrate two−thirds of the way tothe pits of the fru軋

The treatment took about12hours。After thelye‡reatment,the olives were、WaShed with water to

r?mOVeSOdium hydroxidefor about8hoursThenthe oliveswere packedinto2−L glass bottles>

coveredwith8%sodium chloride solution,and allowed to ferment

Samplesofbrine(5cc)weretakenwithpiっettesfromthelotsoffermentingoliv由,WhichpreviousIY

hadbeen thorIOughly agitated by rollinq and shakingtoinsureuniform distribution of chemicalcon−

stituents,dilutedto25cc with distilledwaterina25−CC VOlumetricflask,andused foranalyses De匂eTmin如ion of p問bft恥ebTine so抽tion、t)HValue of thebrine solu七ionwas determined di_

工eCtlyby七estingtheoriginalbrinesoIutionswith p壬壬testpaper(ToyoFilter PaperCo,Tokyo)

De電eTmina色ion of色o電alacidiiy.A5−CC SaTnpleof the dilutedbrine solutionwaspipe七tらdinto a50−

ccErlenmeyer flaskandtitratedtothe phenolphthalein end pointwitho・OINcarbonate・freesodium hydroxide solution‖Thetotalacidity,eXpreSSed as grams oflacticacidper10〇cc of brine solution, was obtainedbymutiplyin望the nu血ber ofmi11ili七ersof O‖CIN sodium hydroxide solution usedin

titratioヱ1by Cl..09

De竜ermirla毛ion of sodium ch!oridecontem色。Al−CC Sam‡)】e of the diluted brine solutionwas taken withpipetteandtitrat?dwithoIIIN silver nitratesolutionin the presenceofafewdrops of10%

potassiumchromatesolutionuntila slightpermanentredcolorappearslThesodiumchlorideconte−

nt,eさこpreSSed as grams of sodiumchloride perlつO cc of brine solution,WaS Obtained bymultiplying

the number of milliliters of O.ユN silver nitrate solution usedin titration by5Ⅹ0。585

Es電imation o電■redllCingS嶋arS..Al−CC Sample of the dilutedbr’ine solution was tai【en,deprotei−

nizedby addingwith9cc of;LO%trichloroacetic acidsolution,andused for the determination of reducingsu.gars‥The HA(首EDORN−JEZ(SENmethod(19)wasemployed′rheresultsweIe eXpreSSedas gra−

ms of glucose perlCO cc of brine solution

DeteTmination oflaetic acid eonten七.Al−CC Sample o壬the diiuted brine sollユtion was taken and used for the determlnation oflacticacidafterdeproteinizingwith tIichioroacetic acidar3ddesuga工Iing

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Vol.フ,No.ユ(1955) 95

WiththemiⅩture Of copper sulfate and calciumIlydroxide.,The determination was made colorimetr−

ically according to BAREER and Sun‡MERSON(20)

Sampling was made at3rd,5th,8th,i3th,2Cth,2プth,4フth,66th,and85thdays of fe軍mentation and the arlalyses were completedin 24hours after sampling at each time of $ampling.After・Sampling WaS done,aboutユO cc of′8%sodium qhlo工ide solution was added to eachbottle to fillit up.

RESUむTS A:ND D王SCtTSS‡ON

The analyticalresults were summarized graphicallyin Figurel.Jn TableI,the temperature va− riationint・he fermentation roomwas given,forthe variation of the roomtemperature,under which

the expeIiments were performed,SeemS tO be verylmpOrtant tP COnSider the analyticalresul鴨 TableI.Temperature Variatiollin the FermentatioIIRoom

Temperature Stage of Fementation(Days)

(O Cい)

C・−3 3−5 5・−8 8−ユ3 13−20 20−2フ 2フ−47 47−66 66・−85

Maximum 18.6 20..5 ユ9小C 17..0 ユフ..5 ユ.4。5 ユユ..0 8“5 14.5 MinimuTn lC.8 12..5 10..9 9..0 5.0 6.5 1.0 ユ.5 1.5

The results shownin Figurelarein Close agreement with the descrlption of VAUGFN,eiallr(2),anditmay be seen that the fermentation of the sample proceeded in anom]almanner”Very active chemical Changes were observedinthe early part of the primary stage of董ermentation,andthe importanceofthisstagein the whole stages Of fermentation willbe easily understood Asindicatedin Figureユ,theprimary$tage SeemS tO PerSist for7to14 days at room temperatureinthis countryasintheUnited

States..During the七ime the bTine solution reached nearly the stabili2:ation point;the reducing sugars became available for’the bacteriainthebrinesolutionandthenbegan to decrease,COincidently,there were 一次OUヱUS将観︶巴亀コS穿−U苫“貞 ㍍∴︵宅一レ一、菩〇↑苫一“p叩レくUl︶Uq− U lO 20 30 40 50 60 70 80 Stageof FeTmentation(Days†

Figurel..CharBgeS Of the Chemica!Consti電t!entS i‡l七he NormalBri王Ie So音吐電ion dⅦごingt駄e FeE・mell− tation of Spanish−竜ype Gree王10王iYeS

definite steadyincreasesintotalacidityandlactic acid,andacorresponding dropinpH・・Sodium

chloride decreasedrapidlyinthe firstfive days andthenreachedtheequilibIiumAfter theprlmary

stageof ferTnentation,the pH and the reducing sugars of the brine solution decreased further,and the totalacidityincreased constantly uptoaboutO..25%aslactic acidinmore two to three weeks of fe‡mentation‖The$tageSeemStOCOrreSPOndtotheintermediate stageof VAUGHN,・etal.(2)Inthe

last stage of fermentation,the totalacidityincreased further,but theirlCrea3einlactic acid was COmparatively・Small。I)肌1fOU・z()S,eial,く21)have reported that the normalbrinesolutioncontained only

acetic andlactic acids as acidic constituents.Therefore,itmまy be)COICludeithlt theincr’eaSeSinthe

七つね1a=!i持上1th?1a3t Stユ;eislarg∋1y owingJ=O theincreasein a〇etic acid,and the high】y acid・

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Tecb小Bull一.Kagawa Agr.Con

96

consist mainly of the heterofeImentative、tyPe.h this stage,the totalacidityincreased at

considerablerate,but thepHof the brine solutionpemainedatalmostthesamelevel,i・・en,pH4・2・

The fact$eemS tOindicatetheexistenceof abuffersysteminthefermentingbrinesolution”The

impoItanCeOfthe buffersystemofthebIine solutionhas been demonstratedbyBoRJ30LLA,elal・

(5,6,Z2)inthefermentat、ionof Spanish−typegreen Olives・・Theyhave found thatthe capacityofthe

buffersystemincreasesasdiffusionofthesol一ユtion throughtheskinof the fruit and thelactic

fermentationare proceedir)g,anditsIpaXimumⅥaSShown to beat pH3・フ1−3・8(22〉・Thebuffer

systemwilldifferaccordingtothedifferences ofthelyetreatment,thewashing,the varietiesof

olivesusedthelocalities wherethe;01ives had grown andthe fermentation was per−formed,etC

FromtheFigure,itwillbeseenthatthebrinesolutionhadanintensebuffer・aCtionbetweenpH4・41−

4.0.ThebrinesolutioncontainedO小2%reducingsl増arS ati:heendof theexperiment・Thesubstance

expressedasthe reducingsugaISin血estudiesseemstoinclude nonfeImentablereducingsub鏡anq

ces,Whichmaycontainthebitterglucoside,01europein(23),anditsdecompositionproducts

Fromtheresultsofthepresentexperiements,itmaybe concluded that supplementary sugaris

necessarytoobtain thes11fficientlevelof aciditytoinhibitthebacterialputrefaction

StTMMARY

(ユ).Thebiochemicalchangeswer・eStudiedinthenoImalbIine solutions duringthefermentation ofSpanish−tyPegIeenOlives,andtheresultsobtainedwe‡eSummari2;edinFigurelgraphical1y

(2).Aninten$ebufferactionwas showninthefermentingbrinesolution$aboutpH4・0・

(3)..Itwasfound thattheMilSSionvarietyresultednotsufficient enoughaciditywithouttheadd− itionof supplementary sugars,icり0.4%aslactic acid

(4).Thereseemslittledangerofdeteriorationattheroom temperatureunder200C・・Withoutsp・

ecialcontrolfor preservation

AC:瓦NOWIノEDGMENT

The authorswish toexpress their appreciationstoProflSin,itir・6:KAWAmTRA,Laboratoryof Bioト ogicalChemistry,this College,forhisconstantinterestandmany helpfulsuggestionsinthiswork〃 TheyarealsoindebtedtoT・YAⅡAMOrOand O‖Yos‡ⅠIEAWA,bothundergraduatestudentsofthislabor・−

atory,for their participationsina significantpor七ionof theanalyticalworkinthestudies

LITERATtJRE CITED 515(1951);C.A..,46,フ248c(ユ.952) (6) ‥ βoJ..わzS才一.鋸扉..オブ乍〃β.S≠‖ qgγβ乃り12, 115(1952);C.A.リ4■7,657Cg(ユ953). (フ) :Gγα.Sα.S.γαCβ言おS,3,91(1952);C..A., 47,ユ2692h(1953). (8) :A〝αJ♂・=♂αJS∂Cい β坤α刀../諾s。γ く揮お玖, 50$,497(ユ.95、4);C..Aリ48,14032a(1954). (9) :∫み査d‖,48B,42フ(ユ952);C.Aい,46,922フb (1952). (1)KATAKURA,K,NARASAXI,T,.:Tech”Bull小 励gαぴαAgγ..CoJJ“,6,1(ユ.954). (2)VAUGH2q,R”H.,DouGLAS,H‖C..,GIuLLAND, J.R.:C(ZJ≠/■.Agγ.丘砂5fαリBq互互.678,3(.1943). (3)IzQtJ柑RDO TAnIAYO,A.:GγαSαSツ αCβZねS, 3,79(1952);C..A。リ47,ユ2692g(1953). (4) :乃£d.,4,ユ69(1953):C..A.,48.10251 (1954).

(5)BoRBOLLAY ALCALA,J。M.R DELA,elal.:

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Vbl・フ,餌0・1(∽写5) (10) :属放れ一房腐軌い郎gfcαdα,5,143(1951); C一.A一っ46,5741f(1952)二 (ユ1) :乃£d.,7,231(1953);C…A.,48,4フ22b (1954). (12)ⅤIGUERALo8d,J一.Ma。:A〝dgβぶノ盲・ゞ.・.=押吻. 41,520(1945);C..A.,41,4251b(1947.) (13) ,B乱LA王)ARr,F.A。.:A那加.s γβαJ.s¢㌫ 吋α招いノ菖ざ..ツす衰研..,46Ⅰ享,ユ43(1950);C.A..よ 44,ユ0953f(1950). (14) :∫み才d.49Ⅰ王,317(1953);C小A.,47, 12692i(19声3). (15)MERZARl;A..H..,雨占山yA,J..Sい:点料.ム㌢− g♂搾勧ααgダ〃〝り15,225(1948);CA.卜43, 55ユ1c(1949). (16) :乃は・,16,234(1949);C小A.,44, 9507a(ユ.950). 9フ (1フ) :乃g♂り,け,241(1950);C.A..,45,6687f (1951). (18)ApT,F阜:劫切払通わわ如.γα,20,415(・1951) C−Aリ46,9227iく1952). (.19)HAOEpORⅣ,HりCい,.JEySEN,B.N..:βねd始椚. Z..,ト137,92(1923), (20)BA甲柑R S”BりSun(MERSO2q・WいH”:JIBiol・ Cカβ研.リl由,535(ユ.941). (2・1).、pEⅦOlフZ()Sト丁りG.,SrÅDェ・MA¶,F‖H」・,VAl了Gn甘, R.且:./・Agγ..動odCゐβ肌,1,333(ユ954). (22)B?R甲印ヰY ALCALA,J・・M・R・DE 彪dい眉〝g‖Cゐ♂研リ44,2227(1952). (23)CRtⅧ白s,Ⅵ7.Ⅴ..,AI.SBER6,G.L.:J.A傲肌 Cゐβ桝,5∂C.,56,2115(1934).

オ.リブ果実利用に関する基礎的研究

Ⅱ オブ

の発酵中㌢こ斡ける生化学的変化

片 倉 健.ニ・楢 崎 丁 市

緑異境蔵はオリブ利用法の最・も重要なものの・一づである.従来は全く経脚勺に行われていた塩蔵寄港も,

CRUESS,VAU¢甲,その他の多くの研究により,今計ではかなり科学的基勧ミ確立されて来たように見える・Lかし

乍らそれ等の研究結果の振れから理解される様に、,他国の研究結果声泌ずしも我が国の加エに適用し得るとほ衰え ない. 弼等は,我が酌在来の′方法を出来る長け壷重し乍ら外国に於ける最近の研姦結果を取入れた琴準正を確 立する為に・,先ず正常な発酵過程に発酵液申に筆勢する生化学的変化を追究し次の点を明らか㌢こ・した・ エ)発酵初期妃極めて活発な化学成分の変化が認められ %と兢少し,還元糖が0−ト0・・4%と琴出して来,酸が急激に生贅されて来る・ 2)発酵の中期に於て酸は最も多く生産されるが,PHの低下はそれ程著しくない..更に.発酵後期の全酸度とPH との両曲線の廠向から,発酵食塩水はPI‡4−4湖こかなりの綬腐作用を持っている事が推定される. 3)ミツ㌢ヨソ種を用いた場合は全酸度ほ約0.4%(乳酸として)で,腐敗を防止するに十分ではないから乳糖 が必要であると思われる. 4)発酵後期に.も全酸度はかなり増力托するが乳酸は余り増加しない.凡らく酢酸が主に農産されるものであろう が確認はしていない. 泰実陰に収用したのはミッシヨソ種のみで試料ほ小豆島試験場で生産さた呆契を使用した.東研究を行うに当り 引続き有益な助言を与えられつゝある東学川村信一郎教授,並びに贋験を手伝われた山凍・剖順常に感謝する.

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