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(2) S. WAKITA. ;'2. ￣phoric acid. Measurement. of uranium of uranium. quantity. ･same. to. tate). the. reacted. ･the author. solution,. Was. two. FOf these I_2. Result. :50%,. 95%. 99%. and. The. 25. uranium. for. water. 3000. at the. examine. ho叶. an. The. r･p･m･. quantity. of each. inorganic･. and. in. dried. of the. was. Then. order･. the. cultured-on. mycelium at 24o,. days. of ethanol. of 200g.. Obtainment. to. method. :. of mycelium 6.0) of 80cc. for. 一(Ballg.7o, pH. minutes. acid-organic. obtainment. a.. three. the ace-. experiment･ of RNA.. (1) Obtainment. Ran. ALLEN･s. of phosphoric. the. of. for. solution. by. analysed. kinds. which. this. of O125%. solution in ice-cold placed. was. Adding. acidt5･6):. phosphoric. (215% TCA. reagent. centrifuged. 1-Supernatant. organic. soluble. washed. it. on. of H2SO4･. malt. extract. dried. was. 22l･. mycelium,. the. of. ej(tract. malt. in O･8%. NaCl, For was. -used.. b.. Extract. and. -into powder, washed NaCl lO% by -with -1owish ninhydrin. acid. 1.5g･. pH. on. Some. N/P-1.83.. of N/P-1.98). (2) Nature. of. The That and paper･. reacti叫the the. pentose. for. is, the. this decrease. the. relations. the. activity. the. of this. a. the. enzyme. ethanol. and. diphenylamine. in. obtained. of. reaction,. will. given. the. approximated. pure. RNA. of yeast･. a. the. it is N,13･93%7);. And. have. ether･. of this was. of dialysed the enzyme.. In the. lhours. was cultured丘1trate The丘1trate experiment･. the. (inorganic bladder. phosphoric. a. and. membrane. dialysed丘1trate. had. acid. sulphur. treated. was. gone. as. found was. nearly paper. mentioned. solution･. isgiven. e葺periment. in activity. between. hours. Then. activity. b･ In且uence. ethanol,. (87･7%, 2･5%. RNA.. and. solution was RNA. with powder. be pure. disturbed. through. of this. result. in the. acid. nucleic. washed. of dialyser:. In且uence. a.. water･. used. crude. acid･. acetic. (1 1) containing. literatures was. natrium. RNase･. 'by 20 hours'treatment). and. the. it would. of the. phosphoric acid, which dialysed for certain previously. above. extracted. in. active. :. negative. much. running. When HCl. of light yellowish. 1.669) which. or. Dialysis.. against. was. (2･8%) of light yel-. but. 10%. with. centrifuged･. precipitate H2SO4. 1.32g.. obtained, which, Unmistakably, DNA.. contain. A).. it. biwrette. the. reaction,. treated. was. was. 7.63%4);. result. amine. eliminated･ by adding. 1.6-2.0. The. precipitated. then dried .Land. p,. was. portion to. -mycelium). in. negative. was. acid. nucleic. crude. of. reduced. not. was. Then. 5･6g･. grinded. was. mycelium. in air･. Lastly,. method”2,3'･. diphenyl. the. and. and. dried. reaction.. tits unsoluble was. ethanol. dried. It. obtained･. reaction. nucleic. The. "direct. the. was. powder. of RNA:. re丘nement hot in 95%. enzyme. noticeable hours: of. was. in Fig･. 10.. found. weaker. especially Experiments. dialysis. With. the. in the. after case. have. bladder. been. the. of the made. membrane. dialys上s, sulphur as for and.
(3) Studies. the. on. Crystalline. Substance. Mainly. ColnPOSed. 9. ′. 13. J=･ 8. 草. 12. 首. 7. -r+. 11. 冨` O -1+ h. 3. 車9. B. ql. 4. JO A. ). 5. 讃 %. 2. ト ,ヨ?. f= .ヨ ql hb4 0 亡 1トJ. 8 ▼. /. 呈6 l. A Fig.. ￣Reaction. う･. G. and. RNA. :. =. hours:. sol.. enzyme. 8cc.,. total. Reaction. 5.0.. pH. 24. 0. hours. Oう. (45o) A. s/y:j･ 6. 17. 41. 11･ rFig･ hours. hours. dialysed. bladder. membrane. Dialysed,. ESSS]. through. parchment. result. B).. Optimum. enzyme. NaOH. The. 55.47〟g/cc.). the was. solution. were. 4･2-6･2 4.0. on. analysed. on. were. regard. to. the. in ice. of. well the. Dialysed,. for. Dialysed,. for. optimum. of 2cc.. medium 17. hours. 41. hours. lasted. pH of. at. for to. the. pH. of eight. the. and. result. the and 4.8-6.6.. ribonuclease, kinds. of. pH. a. 6cc.. 20 hours. at. 4.8.. separation. mycelia. the. Fig.. report. after. upper. 12.. inorganic. of. by. 45o,. uranium. in in. of. ap.d (RNA-P,. The. organic-P. is shown. some. lcc.. and. CH8COONa, into. made. adjusted. nucleotides. acid. CH3COOH,. was. mixture and. of nucleic. HCl,. (N/10. And. practised. extract glycerine by KuNINAKA6).. Aspergillus. ○-O. inorganic-P. the. and. respectively.. production. acid. enzyme. of which. this. cooled it,. to. added is, the. phosphoric With. was. sol. 8cc. (RNA 5.0. Tempesol. 8ccリpH. ロn-dialysed. pH. volume of. RNA. A-A. solution. various. total. activity.. ribonuclease. ll.. mixed. reaction. mixture. reagent. That. to. used), the. were. which. A. pH:. dialysed. the. 45o.. :. -●-●in Fig.. given. adjusted. was. a. paper. is. The. rature. a. the. mixture:. 3･2mg･), through. Dialysed,. between. Relation and. Reaction. medium. -. L48. medium dialysed. hours. medium. C:. 糾. 9. Rours. Un-dialysed. :. B:. /. 2･. 15mg.),. 4cc.,. sol.. 畠. activities before. mixture. (RNA. I. q5 的. dialysis.. the. 4cc･. ldrl斗. C. RNase. medium. う. O. ヽー. a. 10,. the. rafter. ▼. 冒 色 ヽ_･ヽ′-_･′. of. 10. ′￣ヽ U U. 5. that OTANIIO),. it. was. and.
(4) S.. WAKITA. 9 8. o/＼ ∫/＼. 6 O. 革う. 6. ￠＼. i; 5. 勺. ＼. 鶴. ＼斗. /へ･-･. 1. う. ー一一○. 0. 2･之5.8. 4.B. 8.9. 7.8. う.名6.6. o. アH. 10 Mintlt. 12.. Fig.. The. Reaction. sol.. justed. pH. Reaction. Acid. pH to. the. a. seek. pH and. will. repeat. curve. quantity water. E;≡]. the. and. of veronal-buffered with. a. condenser.. the. activity,. of. re丘nement containing solution. After. the. (45o)I. most. and. If. enzyme･. the. enzyme,. result. in various after. unstable it takes too. pH･. studying. on. will show Then the the. basis･. enzyme.. 30cc･. (M/35,. 10, 30 and. (RNA-P. organic-P. pH,. enzyme. the a. 20 hours. soluble. optimum. experiment of. buffer. Inorganic-P. the. of. activity. 2ccリen-. sol. veronal. 6,0),tota17cc･. hours: Acid. of the. activity. above-mentioned. A且ask. the. at. stable. RNA. (pH. after. solution･. 〃g/cc･)I. Reaction. weakest on. not the. of the. concentration c), Heating:. most. the. whichgives. stability,. author. sol.. organic-P. is found. enzyme. the. long. soluble. 2.5cc.. activity. enzyme. 2･5ccリM/35. sol.. 57･88. Inorganic-P. The at. total. the. mixture:. zyme. 55147FLg/CC･)I 20 hours (45o)I. hours:. ○-o ＋-＋. (ad-. ribonuclease. of. Reaction. CⅢ3COOB, 3cc･,. NaOH). and. (RNA-P. cc.. HCl,. sol･. The. heating. 2cc.,. sol,. buffer. N/10. with. CH3COONa 6. RNA. 1cc.,. 13.. Fig.. pE.. various. mixture:. enzyme. activity. ribonuclease. in. 6O. う0 eS. solution. pH. 6･0). 60 minutes,. ofenzyme was. 5cc.. treated of. the. andthe. same. in. boiling. a. mixture. was. tube to 2cc. of RNA (the last conadded solution in a test respectively, case Each was RNA-P 57.88pg/cc.), and then stopped closely.. sunk centration, treated in a thermostat for 20 hours at 45o, and together reagent･ with uranium The in the was inorganic-P analysed･ upper solution Then and organic-P. pipetted. in Fig. 13. result is shown is in close connection Heat resistance with a pH of the of enzyme was is strong ln This the former experiment aCidity･ and, generally out. at. pH. 6.0.. solution,･ carried.
(5) Studies. The. isolation for. 9%. and. Crystalline. the. on. inorganic-P. of. 60. decreased. On. the minutes･ little affected by heating, boiling for 10 minutes and. was. ･after As. for. a. at. the. of nucleic. recognized. that. more. nucleotidase. than. もoo and. 70o. treated. With. regard. periment,. The. production was. of acid. temperature. than. ･60o, the Heat. resistibility. of. much. by. affected. proved. in. stable. So that ･65o)11￣14,】7). led. to. the. other. at. 60o,. nucleic. -each. the. as. Effect. of. the. of. the 2cc･-. RNA was. poured. the. M/35. tubes,. which. ). at. 14.. Fig.. the. and. Nap. Effect. }. 40. ､---ゾ･･-一. 50. Reaction. the. of. the. on. rature. ヽ--}■■￣. 6O. 70. activity. 2ccリen-. RNAsol.. 2cc., M/35. sol.. tempe-. reaction. ribonuclease. mixture:. zyme. (pH 4･8) 2cc･, 弧32/Jg/CC･). ribonu-. btlffer. veronal. total. 6cc.. (RNA-P. ･. this. in. ■---･一-. 之5. Temper&ture. Reacted. for. 1由b 圧さ]. the. hours.. 20. Acid. soluble. organic--P. Inorganic-P. the. have will probably lasted comparatively. To. ヽ-一■ヽ′￣. 15. riboheat-. from. fact. ヽ■■一∪■■■￣. has. (47%. solution,. ー. mycelium. reaction. added. the. and. test. 5. is. In. isolated. NaF:. into. 軍4. applied,. enzymes.. this. solution. ＼. 60o-. 53･37pg/cc･) consisting. centration,. mg/cc･),. And. acid･. enzyme,. E)･. was. (the last concentration,. about. which The. little inorganic-P. 50o). of. ･case. an. a. cc.-was. and. but. nature,. of incubating. experiment. nucleotide. U. temperature, this. a. ex-. enzyme. pH,. separation. from. cleases. 2. was. comparatively. (the optimum. stable. is heat. nucleotides. 50o,. the. to. acidity14).. are. nucleases. a. the. and. former,. at. heated but. it is generally. from. RNA. organicsoluble at a little higher. 50o.. remained. they. as. of. decreased. seen. And to. of mixed-sol山ion. u'. the. activities. of those. ･86%. cc.. at 40o-50o,. practised. well. which. inorganic-P. of. soluble-P. reagent.. result. treated. when. RNA inorganic-P. 14.. isolation. vigorous.. the. the. great. isolates. cooled,. uranium. Fig.. see. a. deal･. which. 15o, 25o, 40o,. with to. of which,. of enzyme-solution,. left. respectively,. then. of acid however,. minutes. it was that when reported inorganic-P decreased notably,. (pH 4.8), each. solution They were. plugged. for 20 hoursat. ￣were. 10. respectively･. a庁ected. 6. :. 50･32pg/cc･)-consisting. were. for. form･er15).. the. temperature. -RNA-P veronal-buffered. boiling. degrades. ribonuclease. the. unstable Optimum. D)･. not. by. =. has. of. was. acid. the. but. stablel卜叫7), is. isolation. the. and. Composed. production. 37%. minutes,. KuNINAKA6). ribonuclease,. 5･5-6･5,. pH. degradation. P. 90% 60. 22%. to the. contrary,. a. Mainly. Substance. mixed of. the. relations. 6cc.. of. enzyme. solution. M/35. (the or. veronal-buffered. (the last Nap. the. stability. of. long.. solution the. with. concentratior),. last. RNA-P. con･. Taka-diastase solution. (0.25 (pH. 3.3×10-2M).. 4.8) And.
(6) S. WAKITA. then. the. mixed. brought. After. at 45o.. the uranium. with. is glVen. in. In. other. ラ リ. DiAStaSe. the. mycelial-. to. the. RNA. 2cc,. 6cc･. the. abワve. for. 匡5] P. Acid-. 3.3×10￣2M) hours. 20. incu･. and. organic-P. author. inorganic. the. moving. found,. was. phosphoric. had. acid. when. to. the. 〃acid. an. enzyme. of NaF,. addition. lost but least. two. in. part. nucleotide5), The. types. above. Nap,. and. was. of. of. experiment. be. deal of. great. in the. seen. the. with. phos-. cultured re-. of. object. it. MgSO4,. by. affected. NaF,. will One. RNA･ not. presum-. will. and not. con丘rmed. led to the. conclusion. degrades by. to be. dialysIS. by a. to. RNA･. affected｡. mono Or. or scar-. fungl. the. RNA. from. acid. its activlty. that. in. found. affected. RNA. was. remarkably. was. acid. heating･. dialysis, from. acid. be. phosphoric. loses. was. by. treated. phosphoric. stable and inorganic. isolates. enzyme. a. was. phosphoric. therefore,. enzyme,. by. not. previously. of inorganic. is heat. other. of. organic. soluble. decomposition. which. the. isolation. solution. the isolation. the. RNA･. nucleotidase”･. uranium. 1abile, is obstructed Though. by. the. isolating the inorganic of enzyme The of the enzyme,L optlmum PH was 5･8･ As its RNA-P nearly. activity. decreased,. the. with. isolate. of. inorganic.. decomposing. acid. to. culturedr. mycelium. varied. velutiPes. phenome-. in the. acid･ the. that. remarkably. obstructed. was activity ably belongs. of. previously. medium. phosphoric. relations. close. the. therefore,. acid. having. dialysed. explain. the. out. carried the. ribonucleases. -phosphoric As there phoric. medium,. to. that. medium collybia. was. was. experiment. synthetically non. Inorganic-P the. ribonuclease･. on. not. on. e庁ect. obstructive. and. 45o.. at. soluble. (pE. Nap. mixture,. (Nap,. added bated. The. 53 37〃g/. (RNA-P,. cc.). *To. an. Consideration.. 2cc･. sol･ bu庁er. veronal. 3.. 2ccリCul-. sol.. diastase. or. (0･3皿g/CC･),M/35 total. has. recognized. activity.. mixture′:. 4.8) 2cc.,. it is. occasions,. most. nucleotidase. the. on. turefi1trate. the. to. organic-P.. NaF. that. fCiTttruartee Effect of Nap. 15.. Reaction. NaF.. 97%. of. an-. showed. 80%. soluble on. ユ. J. -. ribofluClease. take. NaF. words,. soluand solution for Taka-diastase, the compleof phostion of the isolation of inorganic little･ NaF And sho甲ed phoをic acid. to in order produce acid obstruction,. Fig.. At. result. tion. -_､土j′. cely. The. 15･. culture. う. It. treated. was. reagent.. Fig･. obstruction. 苛4. it. cooled,. processb. for 20 hoursb. incubated. was. out. this. solution. Which or. oligo六. addition This,. ofー heat-. dialysIS･ the. nucleotidase,. this enzyme. is. a. result. nucleotidase. of ;16)-.
(7) Studies. from. and. this from. most.1y 0n. we. then,. as. the. V. varied. a. isolatedr. be. will. from. polynucleotide1. mycelia. Method. of the. (1) Obtainment b.. Medium:. c.. Culture. the. Mycelium. Periodically, ing. 0.9%. and. a. 0.19g. CHUNG's. into. washed. of. carbohydrates.. solution b.. KOH for. treated. 7.5cc.. of. water.. with. into. a a. three the. with. 30. centrifugal. refrigerator times. with. precipitate, water,. then. tube at. washed into 50cc. The in. minutes and. were. author. gathered__. and. for. in. dry. 30mg.. var.. was. cultured. Erlenmeyerjiask with. water. was. was. weight). a. then. of. 5o. for. 15cc. 10. with. with. then. minutes, water. with. in. a. by-. morter,. and.. trichrolo. acetic. cold. loc°. of lO%. a. and. the. centrifuged, And. centrifuged.. solution author. precipi-. the. upper. water.. residuewassuspendedina After boiling water.. 5cc. it. cooling,. The. precipitate into 20cc･ with. centrifuged.. of 30%. KOH. was. of 0.1N. NaOH.. dissolved. it by. Adding heating･. 1-2cc.. a. It. was. and. diluted. was. with twice･. washed. was water･ made solution IOcc･ KOH Fehling's A extract of solution: a 4cc. Fehling's tube, added with of solution, The all night, and precipitate centrifuged.. analysed.. rest. separated. by. the. contain--. The. chitin.. crushed. upper. a. on･l二. 40o.. at. tested.. then. and. tested. 67. velutipes. washed. filter papers. mycelium. twice. water,. The. carbohyd-. :-. The. extract:. Precipitate. c.. below. centrifugal was kept. made. was. air. (about. mycelium. was. which. in. of. It. tbat〝. carbohydrates.. mycelium. 200cc.. a. mycelium. of between. dried21). a. (TCA).. acid tate. extract. malt. as method19) TCA extract:. a.. the. analysed. The. of mycelium: in of 40cc.. obtainment. were. the. of. of. the. max-. supposed. contents. Thus,. uelutil,es 67 and Collybia (Ballg. 9o, pH 6.0). extract. certain number dehydrated. of the mycelia (2) Separation. the. its. after. it is. appeared, in. extract,,.. malt. medium. Collybia Malt. NaCl,. of the. mycelium.. :. of the. surface. have found. cell membrane. divided and. and. acid; RNAー_. of. experiment.. of. a.. also. nothing. Mycelium.. surface in the. accumulate. should of the. periodically,. the. on. be. phosphoric or. in. Carbohydrates. phenomenon. maximum--. days.. 90. cultured. these. components. chief. for. little. that. its. after. inorganic. lSOlates. of. of which. of且uctuations. cultured. 1.. was. Since. mycelium,. recognized. was. Fluctuation. components. quantity. rate,. varied. cultured. mycelium. imum-growth18).. to. r. acid. phosphoric directly. may. gradually. it. mycelium. Periodical. inorganic. the. and. describedl),. in. the. of. itself. already. The. RNA. the. words,. found. Was. in. thoughit. decomposes. growth,. a. nucleotide,. Composed. OCCaSIOn.. In other. a. inorganic. that. mono. Mainly. Substance. conjecture. can. a. Crystalline. the. on. was. pouredし. agitated, was. of made. 2N. left. washed H2SO4. into. lOcc..
(8) :8. S. WAKITA. d･. Acetic. 7cc.. ￣in a. After. acid. of 2N. acetic. it. cool,. The. extract:. was. acid,. centrifuged. and to. solution ￣total upper ma-de e. Insoluble carbohydrate. acetic ￣H2SO4, and. acid. ￣then. into. ･(3) Quantitative Every. CHUNG's 2.. 1ium. were. was. tents. of. which, fraction. was. in. by. analysed. decrease. days･. were. TCA. its its. progress. the. conan. contrary, younger. age;. of One-third. extract. in. the. On in. one. varied the. mycelia. 8.uctuation. with increased gradually.. KOH. or. the. myce-. two. varied. mycelium,. any. culturing. in. in. carbohydrates In the normal scarcely. the. stage,. the. and. insoluble. the. of. ex-. fractions･. of. qualitative 24o. at. divided. was. to. added. into. in. them. the. reaction. 30 minutes to. ￣were. negative of TCA ･extract The result of. the. these. the. glycogen. no. and. and. times.. several. biurette. the. 皿ethyl. pentose. pentose. that. uronic. acid. are. the. the. (Table. actively. there in. Only. reaction.. reaction. indicates. experiments. ethanol. 3 hours. reaction, nynhydrine Fehling's All solution. reduced of HCl Fehling's reduced They of solution･ the. and. showed. left for. above.. not. 3%. a. resorsin. 90%, 90%. the. as. carbohydrates. by. the. incubated. mycelium. varied. various. washed did. and. the. of. concentrationof to. with. KOH. and. decomposition∴no. the. were. iodine. for. 3g.. reactions:. precipitates tcentrifuged. were These fractions negative. _reaction, them boiled. was. the and normal Figs. 16 and 17.. in. was. found. The. the. and. carbohydrate.. was. days. fraction. in the. total. there. was. to. acid. General. -for 30 Ethanol. on. in mycelium.. weight･. component. came. acetic. extra.cted. ether, to it, agitated,. added. carbohydrate. is shown. the. and. extract. Characters a.. days,. mycelial. mycelial. by. tract. noted. result. 60. abundant,. of TCA however,. amount. (2). for the. especially. Was. residue. dehydrated. and. ethanol. cool.. carbohydrates. The. analysed.. of. in. The. :. eXtraCt). of carbohydrate. method,. In incubation about. the. then. and. experiment.. above. 70%. minutes.. method19).. of the. the. H2SO4. (75%. above. (1) Periodical且uctuation By. 30. water,. with. suspended. for. water. twice. washed. was. of carbohydrate.. the. of. anthrone. Result. with. boiling. a. KOH. with. 20cc.. water. analysis. lcc･. in. 70%, 99% washed with A 5cc･ of 75% H2SO4. was. weighed･ 15cc.. made. extracted. treated. and. was. 'by. residue. no. its. or. protein. above. 10).. carbohydrate. fractions. b. little ing and. Paper of. water･. BaCO3･. partition No.. the. A. partition. chromatography: fractions carbohydrate. above. All. the. cases. The丘1trates chromatography･. 51 (40×40 cm･), and. of this were. for the. were. experiment. is, for. developing. of. treated. and. in. condensed. That. 2%. the. for. WaS. one. neutralized. vacuum. and. filter paper. reagent. H2SO4. were. was. used. to. added. hour. in. a. a. boiレ. Ba(OH)2. with tested. by. used. T6y6. n-butanol,. paper. roshi gracial.
(9) Studies. Crystalline. the. on. Substance. Composed. Mainly. 50 占. ヰO. 80. g P. 5. ･r+ ▼･･+ ￠. ---------I--O1------￣￣-￣◎. 7β. ￠ i. q5. O. L 甘. A 一言. Bl. 30. q】. A i O. ,J). .Jj ). ∫￠. LJ. q5. tH. O. O. 乍0. 式20. †･+. (也. ヰユ. ○. ?. 30. 10. i2Q. ラ. [0. 2 -. --A-. 0. -. -i-_▲. 60 days. cqlもure. 16.. (1) (2) (3). Non-precipitate,. by. the. the. content. normal. Fehling's. 75%. by. in. Fluctuation of. Precipitate, Extract,. ----. LIO. 2O. Fig.. 1. Fehling's. carbohydrates. mycelium.. sol,. H2SO4 by. of. sol･. (4). Extract,. (5). Total. (6). Extract,. by. 2N. CH3COOE. carbohydrate by. lox. TCA. L＋0. ゝ＼. う Pro. ー--○---____. ∼ BO. 買う0 ･lヰ l･+ ￠ O. ＼4＼. き 20. B. ∫○. 4) ∼ a トI rtl A A 0 .凸. 岳 O. 細. i. rJ q5 J}. ヽ寸 0. /. ぅノ. ●■. a. ＼▲. 30. 0. 冒+. 之0. 1O. LO 2. __----●. A. ･････-. 0. 60. 斗O. 20. days. Ct11ttlre. Fig.. 17.. Fluctuation of. (1) (2). Precipitate,. (3). Non-precipitate,. Extract,. by by. 75%. in the. Fehling's. the. varied. content. sol･. H2SO4 by. Fehling's. of. carbohydrates. mycelium.. sol.. (4) (5) (6). Extract,. Total Extract,. by. 2N. CIi3COOH. carbohydrate. by. lO%. TCA.
(10) 10. S. WAKITA. Table. lO･. Reaction. Color. Biurette. reactions or. the. Aniline. reaction. 10%. TCA. ex.. 十. KOH. ex,. ＋. Naphtho-. acetone. reSOrCln. reaction. reaction. sol.. Non-preclpitate. by. Fehling's. 十. sol.. CH8COOE. 75%. or. by. Fehling's. 2N. Rosenthaler. or. phloroglucin reaction. 30%. Preclpltate. fractions.. carbohydrate. Iodine. ninhydrin reaction. Carbohydrate. of. ex.. H2SO4. eX.. H.Ac,. n-Butanol,. H20:4,. saturated. ′■ヽ. 0. h 4). Phenol,. n-Butanol,. 1,5. 4. saturated. with. F2,0. with. H&0. ′￣ヽ. +.). 普. 丸l 屯). 鈍 色) ＋}. g>之. cd >. A. ･P q5. ヰJ. >. o. 'g. ち. てI ･J てJ O ￠ rd ヰ). ●3 ●サ. ∼ ■-■1. o･G召. ○タ ● ●J. 吋 O ･f+. p8 e) ＋〉 d. 可3. h. e&. o･2. ヰJ. O O. -o:. 03,L[. rJ 0 F; q5. 41. ･P c8. ヽ. ＋. rJ O 亡 ∼.. 4＋ 0. ●. 0. ○. ●. t4. ●. ●3pl く). ＋S. 0･l. 3. ＋ユ. ￠/. =. ,J) l .上I J 月. ●. ○. ●. o. o. 0･q. tM. ヽ_′. h l¥. 宝. Fk:. o. E1. K･. 18･. Paper. 41. 2うー561(12う斗56 partition. TCA. chromatography. 2･ SOX. extract･. Non-precipitate,. x1. 2･ xylose･. (1･ glucose･. control. 1･ lox. 雷 & FLl. 5, 'd. Fig･. 賀. o,古. ●之. o. L9. 也. by. KOH. Fehling,s. 5･. mannose,. by. 3･ Precipitate,. 5･ 2N. acetic. acid. 0.3. fractions.. carbohydrate 4･. arabinose,. extract,. sol･. the. of. 3･. 6. 25LIう. galactose).. Fehling･s 61 75%. extract,. sol.. H2SO4. extract.. acetic with. acid･. water),. aniline. and. The which. futalic. not. coincides. with. solution. seemed. not. (3). for. and. extract was. (4 : 1. water. 5)･ n-butanol. :. the. reagent. The. acid･. lO%. with. coincide. with. of coloring. result. TCA,. 2N. by precipitated the Rf of glucose･ to. (saturated. the. with. in. is shown acetic. Fehling･s The. was. acid,. solution Rf. Rf. of. 75%. butanol. H2SO4,. showed. of the. (saturated. water), phenol. used a Fig. 18.. a. precipitate. mannose,. soluti.n.f the. and. single. which Fehling,良. by. made. part. spot. which,. however,. Every. 300mg.. could. be. con丘rmed. Chitin.. a･. mycelium. Periodical丑uctuations obtained. at. the. irl. above. COntent. experiment. Of. chitin:. (1). was. crushed. and. of. gained. the the.
(11) studies. Crystalline. the. on. Table. ll.. The. b.. was. 4.65. 4.93. 2.92. 2.95. 3.61. 3.ll. by the. ether, 4.7%. tained boiling. by. this. the. washed. with. The. result. was. measured7)･. method. The. residue. 3･8%. by. by. HCl. the. of crystalline. for. small. left. It. it. as. an. Indeed, to･. therefore,. Con-. hydrate･. chrolo. substance. analogous. an. into. evaporatingthere･. found. glucosamine or will be chitin. there. in. in. put. turned. impossible,. was. was. it. was. was. was. substance. it. After. condenser･. substance22). analysIS･. results,. a. using. ethanol It con-. mycelium･. reduced･. was. solution. dried. of the. in it. carbohydrate l% HCl･. by. washed. was. solution. conc･. the. insoluble. the. mycelium,. Then. (2)･. of which Fehling's. bit. above. the. crushing. with. crystalline. the. from. sidering. a. too. was. that. was. content. was. When. solution. days, for several. con丘rm. It. (2)･. method. above. KMnO4･. l%. brownish. its quantity. the. nitrogen, for 3 hours. water. above. After. of chitin:. obtainment. of. the. Ill. gained. removed. in. 4. 38. its nitrogen. then. and. Con丘rmation. fraction. by. treated. and ether, in Table. is given. dish. 4.30 2.78. substance･. analogous. fraction. ethanol. a. 4.23. 4.49. insoluble. and. 60. %. 2.88. mycelium. 40. Chitin. its. Varied. 20. 40. or. mycelia･. 60. 20. %. the. of. tl. Composed. mycelium. days. Chitin. was. contents. Culture. N. *. chitin*. Normal. Mycelium. Mainly. Substance. fraction･. uncarbohydrate. (4) Cellulose･ By ble. fraction. and. And. a. by neutralized for those substances, dried,. dissolved. tion. to. added. were. the. the. in. boiling. a. in 50%. boiled. treated. with. solution. H2SO4･. the. It. for. ether,. and. hours,. 2. water. solution･. wasI. powder but. Fehling's it brought. reagent,. carbohydrates. which. ethanol. reduced. anthrone. the. The. H2SO4. left. with. in. obtained･. lox. of. by. washed. were. were. diluted grew. substances. and. which. was. solution. diluted. precipitates. precipitates,. was. powder. H2SO4. solution･. centrifuged,. whitish. 75%. cold. insolu･. (2)･ an. experiment. easily. solution,, forth. the. show･. Consideration. In. and. bit of. green. characteristic. for. they. even. was. by. The upper centrifuged･ HCl (1:1), where misty. it was. when it. when. and. a. when. scarcely dissolved. 3.. then. and. while,. above. to. ethanol. was. reagent. in the. dissolved. was. adding. after. StiWEIZER,s. gained･. and As. was. water.. with for. mycelium It gained･. done. as. the. treating. TCA. the. the. growth. soluble. fraction. of mycelia. such. sugar23)I. phospho-related was. found. to. As. it. mycelium･ amylose. in. a. have was. mycelium,. as. In. rapidly generally. important. found. were. glucose, the. trehalose,. varied. decreased recognized. carbohydrates. a. extracted. the. that by. oligosaccharide20〉. certain the. mycelium･ after. necessary. carbohydrates TCA. maximum. there KOH. growth were. frac-. soluble of. glycogen. (unprecipitable. the. and por-.
(12) 12. s. WAKITA. tion. by. Fehling's. and Though. the. amylose-type, the. and. solution). acetic. which,. precipitation. acid. however,. by. made. be. to. seemed. were. to. negative. Fehling's. the. of. iodine. the. solution. glycogen-type. be. would. be recognized from not the result of paper could chromatography･ ing that there was from a mannocaroloseB5,26) obtained certain that. and. there. inquiries. a. be. should of 75%. extract. the. was. is. a. and. mycelium A. glucan-type･ 1ia,. of Collybia. but. other. 5,o左of. VI･. If they. hydrate. if the. this,. some. recognized. More. the. aerobic. cular. would. increase.. polysaccharides. former. in. and. were were. latter. the. be. not. produced. chitin. of chitin.. the. of. varied. maximum-growth. of myce-. production. increase. not. in. the. any. the. carbo-. medium. advance. of. and. the. culture in. thar. anaerobic. large. comparatively. comparatively. the. contains. decomposition,. in. produced. mycein. Medium.. would. mycelial. the analysed contents increasing with the. author. In the. medium. by the. medium. in. after for the. used. occurred. in. contained. may. its. chitin,. mycelium little of. chitin. found. polysaccharides. in the. polysaccharides. condition･. a. or. a. of a certain bacterium little less than that. decreased. mycelium. was. found. be. Polysaccharides. notably. polysaccharides. of these. have. any. and. component. was. not. In the. chitin-type. will. extractable. contents. has. cellulose. of. chitin. of. TCA. the. the. cellulose-type,. cellulose=28)･. nitrogenous. content. to be. the. any. deal. great. Fluctuation. by. in. Considering. days･. its nitrogen. decrease. contents. not. (polysaccharides. carbohydrate. And,. more･. a. the. of chitin. above32',. 1ial components,. exhaustive In the mycelium.. a. types-the. has. investigators22･28-30). uelutipes,. consumed. of. Penicillium24),. mannan,. in. mannan. of three. Though. recognized. were. contained. which. has. chitin found of. Periodical. were. 皿yCelium. has. as. mentioned. existence. mycelium. were. type. nitrogen)31),. As. 〃a. mushrooms･. mycelium. not. it. Consider-. glucan-type.. number in. especially. some. the. which. velutipes. the. Carbohydrates. that. reported. Collybia. of. Were. and. did. which. into. made. H2SO4. chitin-type19,27). It. mycelium. reaction.. mannose,. molecular. small. molepoly-. saccharides.. 1･. Method. of the. Mycelium Medium:. Malt. Incubation: of. in. 60cc.. by. hours. in. ethanol. ‥f U⊥. and. ether. Quantitative. 7o, pH. mycelium It. medium:. water,. was. ･. then. in. the. on. vacuum. Poiysaccharides. (pI1 6.0) dried. cultured. treated. was ･. ethanol. 6.0. malt. extract. 24o.. at. nrtl-,----h-l',i-DLtしし1▲uL⊥uしD fJU⊥J. acetous. ice-cold. the. uar.. Erlenmeyerj7ask. of. ▲ヽ/ヽ-'▲tJ▲しLtしIV⊥▲. precipitated. Ballg.. Generally,. prFI,1;∩;十っh.nll. 67. uelutipes. extract,. 200cc.. a. Condensation ▲. experiment.. Collybia. :. on. and. centrifuged H2SO4 after. 40o.. at. tll-1e. irl. after washing. lTlediulTI Were leaving for 3 95%. and. 99%. in order. analysis. of. polysaccharides. :. Polysaccharides. were. analysed.
(13) studies. by. the. 2.. Result. the. on. Substance. Crystalline. 13. Composed. Mainly. method19)･. anthrone. of the experiment. in As there was much Polysaccharides polyextract medium: malt by the general prepared method, (150 mgノ100 cc.) in the malt extract saccharide had been was extract previously the experiment made which of the malt. (1). by. treated. 別trate. was. made. incubator,. 60. of 5g. were 2.4J. to. Taka-diastase. and. the. and. days･. in the. Then. by. adding. it. cipitate, ethanol.. The. ethanol.. Then. 150cc‥. into. 50cc･. the. solution. the. carbohydrates. contains. of 70%. corlCentration. last. the. water, upper. solution. preci′pitate. by. treated they. fraction. a. ether,. anthrone also. were. carbohydrate. contains. contains cipitate by ethanol and. into. and. hardly weighed･. With. reagent･. treated. regard. by. anthrone. the. initial medium,. reagent. at. the. above. in. contains centrifuged By adding. order･. centrifuged･ 30% and ethanol. by. fraction. This. other the. carbohydrate time･. And the. was. in the. same. 81-90% 50ccリand. was. rest. pre-. of 80%. by. solution. it wassuspended to the. case･. concentration. ethanol. in water･. Then. to. the. 50ccリand. precipitable soluble. was. upper. And. ethanol･ 30% and. made. was. The. ethanol･. by 71-80% precipitable carbohydrates in the concentration of 60%, 50%,40%. precipitation. precipitable into made. was. water. with. added. the. in any. the. for. the丘1trate. of. a. kept. water in. centrifuged. in. the別trate. of 90% adding. The kept. was. with. then. and. By. and. precipitate. half. other. concentration. of polysaccharide:. upper. in. centrifuged. the. the. simultaneously into. to. 1/2 of the丘1trate on. cultured. was別tered. ethanol. made. was. 55o. at. of 2･4l･. extract solution of malt for 6 hours andfi1tered･. mixed. boiling,. coIICentrated. Fractionation. a.. a. After was. medium. and. detail, heated. mycelium. the. incubator. obtained. In. Taka-diastase･. The. the pre-. washed water. and fractions, result. is. 79%. of. in F､ig. 19･. glVen. in. polysaccharides. were. 61mg･. per. lOOccリand. う0 与O O. 営30 I-. ?20 10. 0. 8之O-hard)/う0. うト4O. soluble fraction. Fig.. 19･. ”. polysaccharides. before. E22Z. Polysaccharides. after. *. Treated. by. precipitate,. Polysaccharides. by. Taka･diastase. the. of. the the. extract. malt incubation. incubation. 6ト70. 5l-6o. 4l-50. 7l-80. 81-90. EtO‡壬(港) medium*. ⊂コ. (Ballg･ 7o)･ Non･carbohydrate.
(14) 14. s. WAKITA. them. were. 90%. of small On. ethanol･. molecular the. 187 mg･. weight ･molecular b･ Non-carbohydrates:. it. of. was. the. part,. nitrogen. in the mycelium32), pound or its decomposed product.. (2) Polysaccharides The. mycelia. washed incubated land. the. cultured. there. was. in synthetic. the. removing. especially. chitin. or. medium. the. an. amount. the. water-. be. will. non-. of. analogous. com-. chitin. medium. 680cc･. for. of 30cc･. 25. days,. respectively. the medium. mycelium, by. treated. was. In. the. of. an. of nitrogen. in. in. cultured.. (8.5%. precipitation).. extract malt of =ENNEBERG,s38) medium. After. was. part. percentage. non-carbohydrate. Thefi1tration. obtained･. ; the. the. on on. for 90 days･. 150cc･. As. produced. transplanted. and. 6･2･. was. total. 5･1%. was. mycelium. water-insoluble incubation, and. after. content. the. after. 81-. different. each. a. (7･0% of the. therefore,. carbohydrate,. was. medium. non-carbohydrate. insoluble. lOOcc･,. per. in. precipitated. polysaccharides,. There. in. precipitation). be. to. as. small. contrary'various. were. total. weight-so. were. and. was丘1tered the. as. ethanol above and of ethanol-precipitable was polysaccharide obtained. a･ Separation and measurement : Having of polysaccharide added water to the preclpitation it into and then made 50cc. The author treated the solution by And was ethanol･ each of its polysaccharides fractionized and analysed. The in Fig. 20. result is shown. ･90%. 7 6. gう. 買ヰ 盲 i:. ぅ. H&0-hard(y. 30. 5ト50. soluble frac Lion Fig･. Though `in. these. the. malt (Figs. 19 and. lCharide 2.9% of b･ Table. 20･. That. produced. it in. the. Nature 12･. All. in. those is, he 911%. synthetic. synthetic. medium.. looked. synthetic medium former was less than. noticed. in. of mycelial. another weight. experiment. in the. in. those. malt. that,. like. those. the. latter. polysac-. extract,. but. is. shown. only. medium.. of polysaccharides:. Nature. the. these. sugar. ,耳tQ.tt(%) a. in the the. 8l-9O. 7ト8o. I)y in. produc呂d. polysaccharides. had. 61-70. _6o. Precipitate Polysaccharides. extract,. 20)I. 5l. composlng. of polysaccharides polysaccharides. was. as. tested. for. in their.
(15) Studies. Table. Crystalline. the. on. Polysaccharides. 12.. Precipltate. Fehling's. EtOH. by. 71′-80. by. 61-70. ”. by. 51′-60. ”. by. 31′-50. by. 30. ”. the. synthetic. medium･. Component. ＋. ＋. ＋. 3′ヘノ4. ＋. ＋. 十. 4′)5. ＋. ＋. ＋. 8′･)9. ＋. ＋. ＋. 13′.-ノ14. ”. ＋. ”. ＋ ±. fraction. hydrolysed. by products in the 皿OnOSaCCharides by. culture. ･of lOcc･ glass. fluctuation of. poured for 40. ℃ubated. tubes. was. into. test. days,. one. and. 150%, in Fig.. tubes,. each. fourth. of. them. Periodically. of. into. they. the. 50cc･,. the. then･. and. treated. as. fractions, The. fractions･. test. separated･. Treated. water-hardly-soluble. a. with. in. were. in-. were. medium. medium. polysaccharide:. insoluble. ethanol. extract. malt. When. polysaccharides. precipitable. divided. into. concentrated. and. the. the. on. into. sunk. were. medium･. extract. cultured diameter･. in. 2cm･. measurement. became precipitation 66-80% 51-65% and. the. was. power. reducing. in malt. polysaccharides were The mycelia. of polysaccharide:. Fractionation. c.. The. polysaccharides･. of. obtained,filtrated, 80% of the ethanol. above,. hydrolysIS･. after. of. mycelium:. rod. Obtainment. b.. as. conditions. Periodical. averaged number of from the ratio between. obtained. was. method･. (3) Productional a.. and. the. and. chromatography,. polysaccharides. SoMOGYI's. ｡measured A.. paper. of before. powers. reducing. condensation. (glucose). Water-hardly-soluble. the. Molecular. saccharide. reaction. sol.. 15. Composed. Pentose. by. reaction. Mainly. in. produced. Iodine. Saccharide. Precipitate by 81′-90%. Substance. above,. 30%,. result. is. 31shown. 21.. by 60-80% ethanol part precipitated molecular on the decreased ; the contrary, part respectively 一and the water-hardly-soluble increased 50-60% the with one by ethanol molecular precipitated middle days･ Though the middle molecular polysaccharide advancement of the culture decomposition the higher be by moleof to the be 雅ay produced considered lead to the one in the medium, the will medium result of this synthetic cular. In. that. conclusion ･of. words,. other. the. increase. of these. is. polysaccharides. by. autolysIS. in the. medium. caused. mycelium･. when. the. -increased34,35). was. mycelium in. quantity,. precipitable. cultured. in. the. be. medium. polysaccharides. anaerobic, notable. inactivity will. polysaccharides. polysaccharides. was. which The. polysaccharide･. :soluble. cular. the. low. the. due and. of to the. especially the the. in. the. of 66-80%. decrease degradation86). autolysIS. water-hardly-. Of. a. of. higher. mycelium. ethanoI mole-. though.
(16) 16. S_ WAKITA. 90. 80. ＼･＼. 70 ○●. 6o. U. ⊂) ⊂) r-. 盲. 50. ￣--●. E LIO. =). う0. O ●. ”:i4. __-----. ●一--一■￣. '.. 川). ′. /. 20 ′′. _一▲. 10. く.A空車≡ラ. (&J 0. 0. 25. 40. Culture 21･. Fig･. Aerobic. ●-● ＋1----＋ *. in. culture. Anaerobic. Before. (1). Fluctuation. the. culture. EtOH. content. Ballg. fraction,. polysaccharides. the. of. medium.辛. 7o. 30%. (2). 51-65%. (4). precipitate,. of. (on the medium) (in the medium). it was. culture,. Water-hardly-soluble. 50%. a. 60. days. EtOH. EtOH. (5). 31-. (3). precipitate,. precipitate,. 66-80%. EtOH. preclpltate.. the. mycelial This. vitality. displayed. the. under. weaker. varied. to. tendency. a. become. will. that. means. mycelium, into. break. condition.. anaerobic a long. after. the皿edium,. period. especially. of. culturing♪. under. anaerobic. condition.. for. B･. Effect. a･. Culture. 25. floated in. Bed. a. half. the. diameter,. of. in. days. test of. tubes,. them of the. cultured Obtainment. the. ethanol was. above. above was. the. of. 2cc･. half. product. in diameter,. magnesium, in the same. of polysaccharides.. mycelia. of lOcc.. the. on. the. of in. test. about. And. and. washed medium. tubes. medium.. extract. malt. author. HENNEBERG･s. to. the. the. washed several cI Separation and. the. in mycelium. polysaccharide. times. experiment,. about. and. other. experiment,. added. culturing. surface. glucose. upon. After. modi1.9cm. in. each. them. of. days.. 60. b･ in. each. on. sank. and. magnesium. mycelium:. contents. for. was. and. of glucose. mycelium. medium, by 90%. was. 90%. and. dried･. As. medium:. On. in concentration,. the. other. and. the. shown. hand,. some. preclpitation. ethanol.. measurement. polysaccharides. of. in. polysaccharide:Asshown were. SOX, 31-50%, 51-65%, soluble polysaccharides, insoluble The is shown result polysaccharides･ in Table As illustrated 13, polysaccharides. fractionized 66-80% in Table were. into. and 13. water-hardly-. 81∼90% and. produced. the. ethanol-. Fig. much. 22. more.
(17) Studies. on. the. C. A. Crystall′ine. Substance. Mainly. 17. Composed. LLO t4 4) ∼. ql. ■l pt). ･う0. &h. 0 Jl. 岳 ■l I･づ. ＋q1. 20. 0 }l O Jf 〟. 10. o ･■〉. ヽた. 0 A. a. B. A. C. う0. B&0-hardly soltlble fr&ctio71 22.. Fig.. 8. C. A. 5l-50. C. a. !l-占5. アreeipit&te.. Effect. of. glucose of. and. medium medium. lacked. for. glucose. C:. Eenneberg's. medium. lacked. for. glucose. Table. 13.. the. (on (in. culture. Effect of. the. of. the. on. magnesium. Henneberg's.. culture. C. a. 81-90. production. polysaccharides.. Henneberg's. Anaerobic. A. b3苫tOI王(A). B:. 匡≡]. a. B. 占6_80. A:. Aerobic. A. and. magnesium. medium) the. medium). culture. the. the. on. conditions in. polysaccharide*. production. medium･ Polysaccharide. Saccharide･ Method. Medium. Ⅱenneberg's. (control). medium. Lacked Lacked and. *. glucose. for glucose magnesium. Polysaccharide,. under on. for. its. surface, it･. quantity. 18.3. 1.8. anaerobic. 12. 38.0. 3.6. aerobic. 93. 70. 19.7. 2.1. anaerobic. 94. 26. 22.3. 2.4. 17.3. 1.8. 25.3. 2.6. 97. 03 100.. anaerobic. also. (mycelium). 00. condition. but. cc.. mg/100. (medium). 106.. by. also. in. 90%. the. much. 07. EtOE･. than. medium in. more. the. is generally absence of autolysIS in the grown nourishment, polysaccharides In duct this way the product of autolysIS. in. cc.. mg/100. 100.. aerobic. and As an. the. aerobic. precipitated. anaerobic. to ratio mycelium. in quality･. In. other. in. the. case. of aerobic. of glucose. existence. the. stimulated･by medium. will. not. of polysaccharide wards,. de丘ciency. merely varied. polysaccharides. culture･ in the. than. be not were. of. a′'pro-. only･ pro-.
(18) 18. S. WAKITA. in the. ･duced. low-molecular,. condition,. aerobic. in. and. anaerobic. high-molecular.. comparatively. ･condition, 3.. in. both. medium,. Consideration. From. beginning. the. ☆ere. polysaccharides. disappeared,. they. -mycelia,. of the experiment, found in the malt. on. A went FOf culture polysaccharides, ￣inthese. and. a. ℃hitin. component. time. be. will. that,. was. saccharide. days,. probably,. certain. for. used. quantity. the. of. culturing. increased. polysaccharides. of. the. as was. containing nitrogen be a decomposed will. days. noticed. prわduct. of. produced. than. the. on. cultured. in the malt. into being the. synthetic. substance. It. extract.. attracts. which. for a long cultured It was plain enough. of the mycelium.. products. mycelium. that. in the medium. polysaccharides. was. and. a. cell membrane37,38). autolysed. come. will. ･substance for long. extract. substance. which, the. this, the. mostly the. when. non-protein. of. from. Judging. another. though. and. is probable in the. autolyse be. will. less. medium,. removed. a. medium. new. cultured. the. when. poly-. that. is. medium. renewed. As. regards. Substance. produced to. peculiar. the. ･､substance. had. and. species. a. certain. an. development. mycelium,. young the. on. effect. insisted. was. which The. one.. close. under anaerobic condition. rconsplCuOuSly a large that quantity organism of polysaccharides of the. that. a. that. a. ￣was. small great. that. -noticed On. produced. the. glucose. certain. the. bottom author. the. micro･ decom-. yeast. also the. and. noticed. produced. when. anaerobic. SHIODA42) condition. in the existence of. produced. addition a. to. autolysed. by. produced. The. autolytic. was a. be. would. peculiar. on. of the. case. were. the. in the. was. great. mycelium. decreased. they of glucose, deal of polysaccharides were. the. as. medium. in. part. solution46).. The. formerly. autolytlC. Mechanisms. of. the. above. experiment. to. was. be. di銃cult. Then. he. certain. enzymes. various in. a. the. will. myceliu皿may. SubstanceW.. related. that. make. examined. MgNⅡ4PO4･6H20. of a. crystalline. in the. produced to. in. substance. nuclease43). related. Crystallization. author. MgNH4PO4･6H20. is known. complex. it･. in these. Previously. an producing autolytic is it thought that and. of. is very. concerned. VII.. of. under. not. studied. yeast.. were. that. noticed. mechanism. microorganism. take. and. author. much. by. autolytic. us.. Thus,. be. medium,. the. when. the. of polysaccharides. the. contrary,. top. polysaccharides incubated and. medium. amount in. glucose. the. tells. an. of. in. them. of. quantity. in the. soaked. little. quantity. in the. found. were. cell membrane. the. mycelium. KAIBARA41). position. that. terminalis, of Bacillus NoMURA40) an obtained. microorganisms.. of. species. from. GREENBERG39). a substance, in the spore. such. a. pure. the. reason. substance. mycelium precipitate why. there. in MycelitLm. composed. of Collybia. velutipes45). It. of MgNH｡PO｡･6H20 was. mainly. precipitated. in such. a a.
(19) Studies. Crystalline. the. on. kind. mainly of crystalline substance And led this examination mycelium. 1.. 0Ⅹalic. its pH. and. 2.. acid. varied. A. salt. of the. Mycelium. the. and. 2.. of. Result. the. of. By. It. pH:. cultured. medium. was. acid. extremely. for. quali丘ed. the. uar.. distilled. adding. to. water. was. bymeans. made. a. of. the. ll･5o. were. glass. extract. malt. prepared,. The. electrode.. above45).. experiment.. (1) Production ture. old. of Ballg. 3o, 5o, 7o, 9o and incubated on it.. in the. as. 67. velutipes. was. mycelium. were. in the. 5.8), media. pH. Measurement others. recognition:-. on.. went. oxalic. the. experiment.. incubation:. of. 14.5o,. (Ballg.. culture. following. the. in. MgNH｡PO｡･6H20. of. produced. days. containing. Collybia. :. Method. to. 19. Composed. MgNH4PO4･6H20.. of. Method. the. composed. were. ammoni･a. solution. precipitation 1.. and as. Mainly. Substance. and. of ammonia. oxalic. acid,. and且uctuation. of. in cul-. pH. term.. in medium:. Ammonia. a.. its death and. ; in. a. certain. especially,. result. they. produce. was. produced. its. after. bases47,48).. days. of. the. of. into. decomposed. result into the. of deamination, ammonia. In the. sydowi.. after in the medium. ammonia. by. autolyse. are. product As the. inqulrleS. made. of autolysis of Asp. increased exceedingly. course. ･Collybia velutipes, ammonia Fig. 24 shows the content of. degraded. WooLLEY37,49,50). in the. ･advancement. into. acid,. thingbeginsto. is acted along with its growth22), it becomes As the vlgOrOuSly.. autolysIS. its. and. nucleic. ammonia･. living. a. maximum-growth,. protein. and. acids,. however,. mold,. of autolysIS,. amino. Generally. its. which of. mycelium. maximum-growth･ increaslng. With. the. cuト. lure.. b.. 0Ⅹalic. Collybia. pectedto. produce. extract. -malt the. ･′as. above, acid. oxalic ･tured. for. on. のn.. long. the. obtained. content. of. isshown. pH. of. days. the. ∴. 白_. ち 0. acid. Fig,. last. mediumis. Relation. 23. medium. 25. so. 75. Days. went. in Fig.. 60. 4). 25. studi-. oxalic. will. production a. and. the culture. as. Theresult. as. Treating. author. ェ. 6. medium.. specially,. the. This. when in the. cultured. 7. ex-. out of the medillm Cul70 days45). He incubated. -the mycelium ped increasing. an. was. oxalicacid. was. -the mycelium. as. it. so. producer,. acid. medium:. known. was. velutil)es. oxalic. in. acid. (1) 9o,. Ballg･. (4). and 3o, 5o,. Ba11g･. between the. culture. (2). Ballg･7o,. 11･5o･. the. pⅡ. of. the. days･. (3). Ballg･.
(20) 20. S. WAKITA. increaslng. basic, 2ち. ￣ducts of (S). be. not. /. 16. ○. O. A. }4. acid by. 14 12. E. ,}> /-ーX. 10. of. the. the. will. 6O. 75. acid Fig.. 24.. tion. of. ture. days.. the. concentra-. NH40Hinthemedium. Ballg･. 3o,. Ballg･. 9o,. (1) (4). between. Relation. (2) (5). the. and. Ballg.. 5o,. Ballg.. ll.5o.. conditions. magnesium a. of. therefore,. and,. The. contents. from. substance 6H20, so. cuト. as. has. acid. nium. phosphate･. medium,. and,. substance. study. of. solution a･. and. ammonium. pH,. the. of. of. a. are. important. upon. mentioned. this acid, in the. we. conditions. of salts. Effect of pH. mycelium.. and. mixed. solution. Many the. no-. of oxalic began to. will be potent. oXalic. In view. enough. producing. magnesium. on. production. of crystalline to. the. of. the. three. crystalline. that ammo-. magnesium. in. is produced. to. the. produce. author. ammonium. above. the. recognized5卜59). acid. ofallthis,the. consisting by. of MgNH｡PO｡t. those. with. product. above,. establisbed. ammonium, and di丘cult. in. students. of. corresponding. one. was. ammonium. has. the. crystalmade. a. in. the. phosphate. substance:. 2MgC12＋(NH｡)2SO｡＋2KH2PO｡一2MgNH｡PO｡ A. old. took. compounds. varied. of magnesium MgNH｡PO｡16H20. effect in the. think,. a. in close connection. composition As. in. an. of producing in salts solution.. contents pure. is concerned･. an. acid. pH. in. students46). fact that. produced. magnesium. phosphorus,. a. mycelium. (2) Conditions Some. production. as. down. MgNH4PO｡･6H20. (3) Ballg.7o,. the mycelium. long. oxalic. line. solution-a the. long,. decomposition. when. veluti-. oxalic. NAGATA66). the. de-. autolyse.. the. of phosphorus,. of. goes. a. acid. Collybia. of. one.. after. increased.. activities. before. medium. tice of. D4yS. of oxalic. oxalic.acid. the product. cease. the. 45. 之5. been. pH inorganic. or. medium,. cultured. O. 23.. have. lower. a. organic. ♪es, too,. C. Fig.. S甘IMAZONO51). to. reducing. carbohydrase In the mycelium. 』メ. to. on. adding to. pH. his studies in a certain destroy fungi that, by means. wood. (り. will a. tendency. a. enzyme5ト56).. reported. r一. old. as. decomposition. a. an. acid. endlessly, has. into. made. //I. t〉. to. oXalic. produced. and. protein. of medium as in fall down shown And many Inquiries. ･(.'/. 18. pro-. regard. the. 20. 盲. acid,. mycelium,. cultured. 22. g. nucleic With. the◆ others.. '之斗. decomposed. --------(1) equation. was. prepared. in.
(21) Studies. 且asks.. Crystalline. Substance. was. 1ラ. the. on. A. of the solution pH by dropping NH40H. adjusted. they. when. it.. on. certain. flasks. the. respectively,. a. ど:. pH. were. cork-. 11. The. left all the night. result in Fig. 26 (the is shown result shows ed. and. percentage. b.. Effect. duction. to. of precipitation. deposited. amount. a. at. of. the. :Fr. 9. '. 8 I) O. :. substance. 7. O. 2MgC12＋C204(NH4)2-ト2KH2PO4-. ○. ■■■. ヽヽ. ” ≡. ---(2). mixed. to. was the above preequation in flasks, and as by treating. done. in. 6. correspond-. solution. precipitation above, The obtained. result is shown in Fig. 26 (the resultshows a per-. the. deposited. amount. When the. of. to. of. in salts was. between. already the. Table. tion. and. content. magnesium 14.. that. Relation. Between. Content of MgC12, added. in. Composition. in. 1/2. l.0167. equivalent. B. l. ”. C. 2. ”. 2. 0333. 1/2. B. l. C. 2. *. A. percentage. ”. 2. 0333. ”. 4. 0666 to. thete. are. A. B, after. red. the. ratio in. the. 1()Occ. of. Ballg･. 5o,. (5). Ballg･. 11･5o･. the. important. the. and in. the. existing. compounds. precipitate. and. of. solution. JI. H20. the. the. solution.. J∫. ･. :. substance relations. 1. 3609. ′タ. Preclpト. Pod/Mg. (moトratio). 48. 35. 0.93. 100, 00. 0.94. 109.. 91. 0.92. 49. 57. 0.99. 82. 00. 0.97. KH2PO4 1. 3609. 119. heated･. the. Ballg･7o,. KE2PO4. JI. 0. 7104. and. (3). tate*. C204(N甲4)2. concentra-. medium. (2). crystalline. solution. 0. 6603. l. 0167. equivalent. 9o,. (N‡Ⅰ4) 2SO4. 4. 0666. MgC12･6HO A. in. (g) MgC12･6EO. A. 3o,. Ballg･. in. content. the. between. acid. Ballg.. PO4/Mg. the. 75. days.. ratio. the. Relation. oxalic. of. (1) (4). poヰ/Mg. recognized. 25.. culture. com-. magnesiup. Test. Fig.. magnesium. solution. 60. 45 payβ. the. be. 25. o. in. ammo-. seemed. Relations. content. It. 0. difBcult.. paratively c.. therefore,. (l). /. the. acid. magnesium. phosphate. ♯. 9.25).. pH. wasoxalic. solution,. production nium. a. at. there. salts. to. precipitation. ,. ど. was. of. (2. ♂. ラ. the. centage. /,:. ラ. A. pared. /･}/. pro-. ofoxalicacidon. 2MgNH4PO4. (早). /ノ:ニ. 8.80).. pH. crystalline. 0. Jl. O. ing. 21. Composed. 12. a. attained. Mainly. 29. 0.74.
(22) 22. S. WAKITA. 100. -}>->-'山●. ′･一･. 90. /. 80. /-. 70. /. ′ヽ. 苔6o. 2. 也. ●. IO. td. I: FL. 50. ･rl. 革 L10 Si. Ll. Fi.. う0 20. ●. ●. /. ノ. 10. ○. 5. 7. PH Fig･. 26･. Relation. between. the. (1). produce. the. and. phosphate. (NH4)2SO4. ㌔ammonium in it46). produced. .1!0. To. 誉. 還120. the. 〒芹==. 90. :====... Effect. production. of. Potassium. (2 (3. Ammonium. oxalate. Magnesium. oxalate. to. They. these且asks. was. analysed. (a mixed nesium were. to the. by. adjusted usual. method.. the. a. lOOcc.. equation Ammonium. and. 7.5 The. oxalate,. magnesium. by. cor-. (1) was. above. of oxalic. pH. result. production. of every. oxalate and. mag-. 0.9g.. were. acid. chrolide) of 0.3, 0.6 and to. solution. The. on. substance. oxalate. the. :. solution. solution. and. precipitation,. oxalates. in flasks.. potassium. solution. 14.. of. mixed. prepared. oxalate. a. of. the. Table. Effect. responding. a. on. precipitate･. (1. preclpitation Fig.27. 3.. of oxalates the. between. relations. experimented SO4￣￣ or C20｡￣∴. crystalline. A. (&/1 00cc･). oxa1&te. 27,. 0･9. 0.6. o.5. of. which. in the. in. d･. of. in. ratio. has. is shown. phosphate. contents. containing. 80 o. sure. magnesium. author. (1). よ10e. make. PO4/Mg. .)/. i 1 C. .rlFL･ O O. added. medium.. 出穂hesium was. Fig.. ammonium皿agneSium. the. (2) C20｡(NⅢ4)2. 1んo. ＋ ･r1. of of. pE. NH40H.. result. is shown. Consideration.. According. to. the. experiment,. the. precipitation. of ammonium. magnesium. The in.
(23) studies. began. phosphate a. at. 5･78. pH. of the. will. in. will increase Either potassium. than. a. to. in. magnesium. of. 1. than. increasing. precipitation. Sod--. containing. solution. ln. a-. as. proportion. as. proportion or. that. shown. previously. a. ammonium there is. pH. rise･. a. amount. qualified. it, the. increases,. precipitate. of. (1ess斗. magnesium. ratio. in,the. is. amount excessive become less･ ratio. an. Pod/Mg. and. from･. precipitation. prevents. oxalate. precipitate=. (Table 14).. oxalic. if it is in. acid, the. of. appearance. produced. thought,. therefore,. that. by. produced a. showing in the. the. or. an. metabolism of. and. a. were. irk. WOuld-. excessive. crystalline that. magnesium18) is. which. substance. crystalline. is･. of the substances18'wbich. precipitates. acid. oxalic. there. unless. some. mycelium,. old cultured. It. Mg(NH4)4(Pod)2. that. solution. crystalline･.. Mg(NH4)4(Poヰ)2. as. such. experimented. ammoniac. mycelial. solution-a. little salt. a. (BALAREFF46). in且uence. great. be. must. the･. (mol ratio)･. POJMg-1･2-1･09. mycelium,. there. in it)･ In the. magnesium. old. the. the crystalline substance be produced in a neutral. disturb. (Fig･ 25), will medium cultured In the in the mycelium･ substance. an. crystalline in. substance. are. a. 2詮. Composed. equivalent) in the solution, Pod/Mg 1 (mol ratio) (Table 14), but, if there. molecural. approaches. in. 5･10. pH. --the. oxalate When existence.. into. coming. Mainly. Substance. are in the mediumoxalates is above 5･8, magnesium As a pH of the medium ammoniumin the be if the medium,, exist component salts precipitated,. has. it. and. C204-. author. myceliu皿･. phospbate. a. at. grow. containlng. The. rose･. pH. to. Crystalline. the. on. substance, are. released. composed. mainly. faculty. of mushroom. of. MgNH4PO4･6H20.. Summary l.. Collybia. development,. 2'elutiPes (the. Studying. the. and. sterile. on periodical且uctuation Crystalline descendant.. there. and. the. to. for. trace. a. MgNH4PO4･6H20,. old. its volume. the. of. distinctive. a. of the. sterile They. mycelium･. diminished. was. by. its glassy in air, and surface By at loo° in water･ analysIS♪ qualitative. minutes. minutes. mycelia. observed. in the. years･. several. components. magnesium found. (2×3mm.)I. phosphate, and. author. the. 150o. at. magnesium. and. of silicon･. contaminated. Its. with. chemical one. with. ammonium,. percent. formula. was. of calcium. little. a. identioxalate･･. the. is a product mycelium･ of the autolysIS Of substance crystalline by on medium prepared the 2. The extract malt author cultured mycelia in the Inorganic were at intervals, components analysed metbod･ general and. mycelia was. be. oxalate,. were. system. heating丘ve. much. and. components. heating丘ve. found. was. calcium. The. by. the. descendant,. phosphorus. triclinic. a. one-third dull by. turned. Bed. be. to. about. the. on the malt extract medium cultured between in inorganic difference component. strain. seemed. lost. strain) has. when. the. parent. parent. a. the. medium･. little mineral. In the. component. normal exuded. mycelium from. the. (the mycelium. parent into. strain)･ the. there. medium,.
(24) :24. s. WAKITA. 70. after. -even. 'perature. components. ium･. 3･. The. も･0),and. with increased. these. the. in. and. salt. tem-. cool. (the. mycelium varied 40 days, about such. calcium. into. exuded. compounds. slightly. the. medinto. The. medium.. less than. a. mine-. the. accumulated in. was. which was. mycelium. at. the. for. phosphorous. magnesium in. acid. were. it. mycelium. a. reached Aft占r. the. -pod/Mg. the. they. into. again. by. the. 10 percent. 13･93%,. P. 7･63%, by. (mycelial. the. nuclease. old. optimum. pH for. sensitivlty divided was. inorganic-P. splitable,. of. Collybia on. heating two. the. to. heat. them. Fehling's. lO%. the. young. compounds. to. maximum.. the. decreased. gradu-. mycelium.. Then. culture,. of them. some a. crystalline. the ribonucleic mill, extracted acid barium It was acetate solution. the RNA of the sterile descendant,. by. was old culture丘1trate filtrate, there was much as. prepared activity` NaF･. By. enzyme one. treatment. activity. RNA. NaF･ by. enzyme. The. other and. its. of dialysis, the. of degradable,. dialys上s. By. solution. decreased.. remarkably. the. was. and. an. ininor-. culture. was. and. 丘1trate. stable to heat inorganic-P. NaF･. The. optlmum. 50o.. (the. extract as. in the. producing. mass). culture. interfered. was. were. malt. dialysis. with and. the The. groups.. velutiPes. fractions. NaF,. or. interfered. feeble. by. old. by. in the. releasable. 5･8 interfered. was. into. not. temperature. was. the. culture,. a. system. the. synthetically. the dialysed丘Itrate. treatmerlt,. in. purified. So. ganic-P･. rcultured hydrate. medium.. Applying. l･83.. In. vestigated. 5.. and. In. soluble-P. absorbed. compounds in the increased. mycelium. NaCl,. early. or. inorganic. came. organic-P. compounds the. defatted. acid. phosphorous. mycelium. N/P. decomposition. other. the. compounds. growth,. phospho-. the. were. com-. mycelium. varied in. medium. maximum. and. it into the. organic-P),. is, in. inorganic. abundant. organic. phosphorous. from. in the. the. A7-P. growth,. substance. 4. Grinding. this. to. minimum,. exuded. (RNA). relatively came. maximum. out. returned. culture. phosphorous. residual. That. (Ballg. 7o, pH. medium. mycelial. and. days･. cQmPOunds. and. inorganic. and. ally. the. phosphorous. When. the. In the phosphoprotein-P. were distinctive/fluctuations. by. There. mycelium.. extract. malt. A7-P,. (inorganic-P,. acid-P. the. all. a. quantitatively. analysed. soluble-P. nucleic. almost. on. mycelia. and nucleic acid-P descendants, there. the. and. land. magnesium. incubated. were. But. incubated. were. incubated. author. as. ･sterile. was. when. periodically. pounds 1ipid-P,. the. they. the. ratio. they. of MgNH4PO｡.. ratio. N. mushroom. phosphorus, 50 days incubation,. mycelium. Po鐘/Mg. When. developed.. as. After. the. 24o.. at. culture. the. descendant),. sterile ral. days. soon. parent. medium trichloro. strain) and acetic. were. and. its sterile. analysed. acid､(TCA) 2N. descendant. periodically extractable, acid. were. its carbo30%. KOH. solution precipitable, acetic and extra･ctable, In the normal or there the parent, was mycelium not distinctive di庁erences in the young in any But the varied and old mycelia. or descendant, distinctive there were differences in them. 皿yCelium sterile LeXtraCtable, insoluble carbohydrate･.
(25) Studies. Especially, in. the. TCA mycelium.. 3.8%. of. anthrone N. reagent,. 4.8%,. There. stance.. It. in. reduced. Fehling's. glucose. and. some. which When ･ble. they. were. ous. substance, it will. Perhaps 7.. to. the. 5.78.. The of. was. normal. Mr. for. versity. Of. kind. the. our. advices. this. of writing. ･course. author. or. KH2PO｡,. and. the. the. produced. there. S. WAKITA: G. M.. CLARK,. I. S.a. MoTOE,. A.J.L.. ALLEN:. M.. Agr.. Chem.. IMAMURA: Biochem.. nitr-. reaction･. exist the. at. phosphate MgCl2 and. were. began. 5･10,. pH. oxalate. ammonium. But. normal･. of. to. the. T6ky6,.the. helps. late Dr･. universityand and. Soc.. which. 35, 583. Japan,. Biochem.. I. Agr. I., 34, 858. the. of. by. C20｡(NH4)2,. and. to. were. (1940),. was. Prof･. tO. (1958)･. him. S･. Prof･. Of Harvard. (1961)I 223. the. MgCl2. excessive. glVen. pH. decrease. of oxalate. GR口EN. I., ll, 319 (1917)･ Chem. Soc. Japan, 32,. adding. C204(NH4)2･. 0r. to cordial thanks Prof･ K･ ToGASHI,. have. forms,. precipitation･. obtained. existence. H･E･ they. were. different. time. unless. his. express. temperature in. (NH｡)2SO4. MgNH｡PO4･6H去0,. wishes. in. of pH. thesis.. ScHRYVER:. was. carbohydrate. Re ferences. (1) (2) (3) (4). a. when. "water-hardly-solu-. normal. 皿ay. liquid. mother. precipitation. University. Of the K･ NARITA. the. compound,. the. produced. andvariation. on. phosphate. of. for. precipitate. I.existed. In conclusion FuNAⅢASHI. formation. potassium. of. there. days.. 75. magnesium. used,. extract. polysaccharides in abundance･. In this fractions. production, for. of. solution. malt. were. medium,. were. protein. partition. ethanol･ the incubation,. anaerobicalli, any. ammonium. periodically of MgNH｡PO｡. mixed. a. show. the. on. much. derivative･. magnesium. effect the. produce. iMAI,. and. ammonium. of. (NH4)2SO｡. ･to. acid. not. its. or. the. normally 60% ethanol medium. mycelia, paper. polysaccharides on. or. of. It. H2SO｡･. Though. it with. other. hot. by. observed. before. in abundance･. the composition. _NH｡OH When. weight. 50%. it did. ammonium. to according Precipitates. and. in the. chitin. Condition. for. `tested,. be. tested. were. b).. 6.17%,. 0Ⅹalic. a).. medium. in. produced. N. in. subfraction･. insoluble 50%. descendant. medium. decreased,. were. fractionsりwere. the. incubated. were. precipitated incubated. they. was. sterile. of. crystalline. carbohydrates. polysaccharides in. a. in. readilysoluble. mannose･type. chitin-type,. by reaction carbohydrate Fehling's reduced solution,. HCl. hydrolysed. or. chitin. yielded in the cellulose-type. or. mycelia. it, they. on. grew. When. it.. cultured. and. 13･15%. and. young,. was. to. negative. and. the. or. polysaccharides. mycelium. in. In. the. there. nynhydrin, by con°.. hours. H2SO4,. fractionized. and. were. 3. hot. lO%. ･chromatography. 6. The author medium,. protein. solution･ little mannose. a. was. by. little cellulose. a. was. insoluble. was. to. treated. being. after. It. mycelium.. or. fraction,. 25. Composed. in. 39.73%. was. In the insoluble. the. Mainly. Substance. fraction,. extractable. old. about. Crystalline. the. on. S･. Uniin. the.
(26) 26. S. WAKITA. (5). D･AI. (6). A･. (7). S･ AKABORI:. (8) (9) (10). E･. VISCHER,. E･. BAUMANN:. H･. 6TANI:. (ll). R･. DuBOS,. (12) (13) (14) (15) (16) (17). R･H･S･. (18) (19). S･ WAKITA:. ibidリ35,. CIW･. CHUNG,. W･J･. (20). W･E･. TREVELYAN,. (21). S･ WAKITA:. (等) (23). M･ M･G･. MACFARLANE. (24). W･N･. HAWORTH,. (25). P･W･. CLUTTERBUCK,. MACFADYEN:. J･ Biol･ Chem･,. KuNINAKA:. G･. J･ k6. Chem･. ken. so. 1. ibidリ61, Report R･H･S･. a. P･A･. DuBOS:. KuNITZ:. L･. SHUSTER,. N･0･. N･. Z6LLNER,. J･. A･. KuNINAKA:. J･ Gen･. 24, 15. FELLIG:. Chem”. 535. Soc･. 29, 55 Chem.,. 208,. Japan, J･ Biol.. Biochem･. Soc･. 7, 241. Biochem･. I., 30,. RAISTRICK,. W･N･. 395. (1954). (1936). 1369 (1936).. M･. STACEY:. HAWORTH,. E･. ibid･, 29, 612 RAISTRICK,. GI. T･. (37) (38) (39) (40) (41) (42) (43) (44) (45) (46) (47) (48) (49) (50). D･W･. FuKUMOTO,. T･. WooLLEY,. N･. Bo日ONOS,. R･A･. YAMAMOTO. W･HI D･WI. S. NoMURA: H.. SHIODA,. Chem･. T.. S. WAKITA:. MINE. 35, 686. S. NoMURA:. ibid.,. 30, 237. S. WAKITA:. ibidリ35,. D･. A･A･ M･ D･W･. BALAREFF:. Zeit.. CHRISTMAN‥. STACEYニ. The W･H･. ibid･,. 574. Chem.,. Revs･,. Enzymes. 121, 507. Japan,. 30, 233. 69, 45. (1940).. 31, 421. 114, 85 Arch.. Bact.,. 102, 361. (1957).. (1937).. Biochemリ1,. 319. (1942).. (1955).. (1956).. 173. (1957).. (1961)･ (1956). (1961)･. Physiol･. WooLLEY,. PETERSON.I. 102, 241 32, 303. (1918). (1952).. (SuMNER, JI Biol･. MyRBÅcK, Chem.,. eds･) Vol･ I, (1937).. 114, 85. (1937). SflIMAZONO: 42, 321 (1955). (51) J･ Biochem･, L･ FRANKENTllAL (52) A FoDOR, : Biochem･ Z･, 225, 417 (1930). ibid., 96, 1 (1919). (53) M･ STAlmELIN: 55, 359 (1948). (54) J･V･ BHAT: J･ Bact･, Nature, J･V･ BHAT: 174, 696 (1954). (55) S･R･ KHAMBATA, :. H･. M.. (1954).. anorg.. LASKOWS王くⅠ:. I.. SocI Japan,. ibidリ31,. :. ibid.,. Soc.. PETERSON:. W･H･. 28, 260. Chem.. JI Biol･ Chem.,. EALVORSON:. H･OI. ibid･,. I. Agr.. PETERSON:. J･ Agr･. KAIBARA:. :. WooLLEY,. GREENBERG,. T.. (1935). SMITI寸,. (1934).. : Zentr･ Bakt･ Parasitenk･, Abt. ll. (26) R･ GARZULY-JANKE 15, 537 (1928). Am･ (27) R･CI THOMAS: J･ Botany, Arch･ Mikrobiol･, 10, 515 (1939). (28) KI NABEL: Monatsh･, 29, 1023 (1908). (29) E･ ScHOLL: Biochem･ Z” 167, 68 (1926). (30) N･T･ PROSKURIAKOW: 37, 445 (1939). (31) R･ BROWN: J･ Zmmunol･, J･ Agr･ Chem･ Soc･ Japan, 35, 690 (1961). (32) S･ WAKITA: : ibidリ28, 707 (1954). (33) Compt･ soc･ rend･ biol･, 143, 719 (1949). (34) M･ WELSCH‥ WELSIIIMER: Bact., 61, 153 (1951). J･ (35) H･J･. (36). (1954). (1951).. I., 50, 298. 28, 429. Japan,. Mikrobiol., :. 201,. (1961).. Chem･. H･. (1938). (1938).. 423. (1953). (1953). (1955).. NICKERSON:. Arch･. (1935). (1938).. 1了3, 223. Chem･ 579. J･ Agr･. (1955).. 17, 323. 124, 501. J･ Physiol.,. Am･. J･S･ HARRISON:. ScHMIDT:. 1, 162. (1940).. Biol･. I. Univ･,. Chem･,. ibidリ126,. Physiol･,. Agr･. method), (1948).. 125, 65. ibid･,. KAPLAN:. J･. Imp･. Biol･. J･. LEVENE:. M･. 28, 94. Ky6to. Med”. T日OMPSON:. THOMPSON,. (1934). (1954).. 28, 282. (1924).. Fac･. of. 107, 297. Soc･ Japan,. k沖h6 (The enzymological 176, 715 JI Biol･ Chem･,. E･ CHARGAFF:. ScliMIDT,. ibid‥. Agr･. p･. 956. (1951)∼.
(27) Studies. (56) (57) (58) (59). Y.. NAGATA,. L.W. N.. K.. WINKLER: NEUBAUER:. A.Ⅴ.. EppERSON:. the. on. HAYASHI Zeit.. Journ.. Cristalline. :. I. Agr.. Amer.. Chem.. Chem･,. angeu).. ibidリ50,. Substance. Chem. 321. Soc.. 31, 211 Soc･,. (1928)･. 16,. Mainly. Japan,. Composed. 30, 86,. (1918)I 289 (1894)I. (1956)･. 27.
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