BOHLEY AND FRÖHLICH Reprinted from New Beer in an Old Bottle: Eduard Buchner and the Growth of Biochemical Knowledge, pp. 43–50, ed. A. Cornish-Bowden, Universitat de València, Spain, 1997 A PRIZE-WINNING DISCOVERY OF 1896: BUCHNER PROVIDES EVIDENCE OF CELL-FREE FERMENTATION Peter Bohley and Kai-Uwe Fröhlich The discovery of cell-free fermentation by the chemist Eduard Buchner (1860-1917) is now regarded as one of the great landmarks in the development of biochemistry. Nonetheless, his appointment to the University of Tübingen in April 1896 almost failed. The Faculty of Philosophy had proposed to fill an extraordinary professorship that Fig. 1. Eduard Buchner’s letter of appointment as Professor of Analytical and had recently become vacant by granting it to one of the lecturers of Pharmaceutical chemistry at the University of Tübingen. (Source: Universitäts- philosophy. Fortunately even the rectorate confirmed that a professor- archiv Tübingen, Catalogue No. 126/76) ship of analytical and pharmaceutical chemistry was urgently needed, to assure a reliable and regular presentation of these subjects in the biological course of events, which would never be completely (gesicherte regelmäßige Vertretung dieser Fächer). After some argument the understood… This hypothesis of the vitalists might have dramatically factual necessity of chemical instruction (sachlichen Bedürfnisse des inhibited the progress of research into the processes of life if it had not chemischen Unterrichts) in the Faculty of Natural Sciences prevailed, been possible to falsify it. The vitalists were totally convinced, for and Professor Eduard Buchner from Kiel was finally appointed (Fig. example, that only living yeast cells could produce the much sought- 1) as a capable representative of these subjects (tüchtigen Vertreter für after alcohol from sugar, together with the carbon dioxide that escapes diese Fächer). as bubbles. Separation of this process of fermentation from the living As early as the summer semester of 1896, Buchner taught ana- cells seemed impossible, and had indeed never been achieved. On the lytical chemistry to ten students in Tübingen, and in the winter other hand the heretical idea existed that the “vital forces” were no semester of 1896–1897 he was teaching pharmaceutical chemistry to more than molecular forces, an idea that had been advocated by 14 students and the chemistry of fermentation to 27. He was later Buchner in his lectures on the chemistry of fermentation. His last appointed a professor in Berlin in 1898, but he had made his trail- lectures in Tübingen on this subject were given in the summer blazing discovery shortly after he had taken up his office in Tü- semester of 1898, and attracted students from far and wide: among the bingen. In view of the earlier anxieties of the Faculty of Philosophy, it 32 students (Fig. 2) were W. A. Osborne from Belfast and A. was fitting that it concerned a problem that was of the greatest interest Medwedev from Odessa, and we may also note the name of Otto to philosophers: the miracle of life could not be explained by the laws Dimroth, later famous as a chemist. of physics or chemistry, because a vis vitalis was at work in the 52 51 BOHLEY AND FRÖHLICH A PRIZE-WINNING DISCOVERY OF 1896 Reprinted from New Beer in an Old Bottle: Eduard Buchner and the Growth of Biochemical Knowledge, pp. 43–50, ed. A. Cornish-Bowden, Universitat de València, Spain, 1997 A PRIZE-WINNING DISCOVERY OF 1896: BUCHNER PROVIDES EVIDENCE OF CELL-FREE FERMENTATION Peter Bohley and Kai-Uwe Fröhlich The discovery of cell-free fermentation by the chemist Eduard Buchner (1860-1917) is now regarded as one of the great landmarks in the development of biochemistry. Nonetheless, his appointment to the University of Tübingen in April 1896 almost failed. The Faculty of Philosophy had proposed to fill an extraordinary professorship that Fig. 2. The list of participants in Buchner’s last course at Tübingen, in the Fig. 1. Eduard Buchner’s letter of appointment as Professor of Analytical and had recently become vacant by granting it to one of the lecturers of summer semester of 1898. (Source: Universitätsarchiv Tübingen, Catalogue No. P51h/a6r6m)aceutical chemistry at the University of Tübingen. (Source: Universitäts- philosophy. Fortunately even the rectorate confirmed that a professor- archiv Tübingen, Catalogue No. 126/76) ship of analytical and pharmaceutical chemistry was urgently needed, to assure a reliable and regular presentation of these subjects in thDe ubriionlgo gai cvails ict otuor shei so fo ledveern tbsr, owthheicrh H waonus ldin nMevuenr icbhe icno mOpclteotbeelyr (gesicherte regelmäßige Vertretung dieser Fächer). After some argument the u1n89d6e,r sEtodouda…rd TBhuicsh hnyepro thhaeds ism oafn tahgee dv itaol isptrso mvei gthhta th aitv ei sd praomssaibtilcea ltloy factual necessity of chemical instruction (sachlichen Bedürfnisse des ipnrhoidbuitceed tah ec eplrlo-fgrreees s eoxft reascet arccahp ainbtloe tohfe pferromceesnsetsin ogf lsifueg iafr i.t hOand n9otht chemischen Unterrichts) in the Faculty of Natural Sciences prevailed, bJaeneuna rpyo s1s8ib97le, htoe fsaelnsitf ya ipt.r eTlihmei nvaitrayl imstse swsaegree (tBotuaclhlyn ecro,n 1vi8n9c7e)d f, rofomr and Professor Eduard Buchner from Kiel was finally appointed (Fig. eTxüabminpgle,n thtoa tB oenrlliyn ,l iwvihnigc hy eaarsrti vceedll so nco 1u1ldth p Jraonduuacrey ; tihne itm huec hd essocurgibhets- 1) as a capable representative of these subjects (tüchtigen Vertreter für athftee re xaplceorhimole fnrtos mof sOugcator,b teorg 1e9th86e rt hwait hh athde m caadrbeo tnh ed mioixriadcele t hpaots seibsclaep: es diese Fächer). as bubbles. Separation of this process of fermentation from the living As early as the summer semester of 1896, Buchner taught ana- cells seemIte isd firstimp ofos sallib lprovede, and hthatad ithende initiationed never ofbe theen aprocesschieved of. On the lytical chemistry to ten students in Tübingen, and in the winter other hafnedrm tehnet ahteiornet idcoael si dneoat reexqiustiered stuhcaht at hceo m“vpitlaicla ftoedrc aeps”p awreartue s naos semester of 1896–1897 he was teaching pharmaceutical chemistry to more thtahne myeoaslet ccuelal.r R faothrceers a, saunb sitdaenac et hina ts ohluatdio bne ies nto abdevocated by 14 students and the chemistry of fermentation to 27. He was later Buchnerc oinn shidies rleedc tausr tehse ocnar rtiheer ocfh tehme ifsetrym eonf tfaetirvme eenffteactti oonf .t hHeis last appointed a professor in Berlin in 1898, but he had made his trail- lectures eixnt raTcüt,b winitgheonu t odno utbhti sa psurobtjeeicnt; iwt eshreal l gbieve nna mine dt hzyem assuem. mer blazing discovery shortly after he had taken up his office in Tü- semester of 1T8h98e, iadneda tahttarta ac tsepde cstiuald penrotst efirno mde rfaivre adn fdro wmid teh: ea myeoanstg the bingen. In view of the earlier anxieties of the Faculty of Philosophy, it 32 studecneltlss c(aFuigse. s 2fe)r mwenreta tWio.n Aha. s Opsrbevoironues lyf rboemen Bexeplfraessts eadn bdy A. was fitting that it concerned a problem that was of the greatest interest MedwedeMv. fTrroamub eO ind e1s8sa5,8 aans dth ew een mzymaye oarls foe rmneontte ththeeo rny,a amned othf is Otto to philosophers: the miracle of life could not be explained by the laws Dimroth,h laast elart efarm boeuesn adse af ecnhdeemdi sets.pecially by F. Hoppe-Seyler. The of physics or chemistry, because a vis vitalis was at work in the separation of such an enzyme from the yeast cells has not 52 51 53 BOHLEY AND FRÖHLICH BOHLEY AND FRÖHLICH A PRIZE-WINNING DISCOVERY OF 1896 1 succeeded up to now. In a lecture before the Deutsche chemische Gesellschaft (the German Chemical Society) in Berlin on 14th March 1898, Buchner (1898) proved his discovery in a series of well prepared experiments, with all details publicly demonstrated and elaborated: The alcoholic fermentation of sugar has frequently raised the interest of scientists. There were numerous theories about the achievement of this process until Pasteur’s pioneering works, beginning in the middle of this century, led to the final proposition that there is no fermentation without organisms. Specifically, Pasteur viewed the process as a physiological act closely and inseparably tied to the life processes of yeast cells. Other researchers, however, such as Moritz Traube, Berthelot, Liebig and Hoppe-Seyler, were of the opinion that yeast, in the same manner as it produces a certain chemical substance, an unorganized ferment or enzyme, invertin, likewise produces a substance that brings about the effects of fermentation. Clear as the theory was, however, experimental proof was lacking, because in spite of many attempts nobody had been able to Fig. 2. Tsheep alirsta toef thpaer tpicoipteanntsia lin f oBru fcehrnmere’ns tlaatsito nco furrosem atth Te ülbivininggen y,e ains tthe Fsuigm. m1.e rE dsceuemalrle.ds tTeBrhu eoc hfp n1ue8r9’es8 l.y le (vtStioetaur lriocseft :i acUp tpnhoiveienortrsmiyt eätnthsta erarcseh fiPovr roTefü erbsseionmrg eaonifn, eACdna attalhyloteigc uauel n aN-nod. P51h/a6r6m)acecuotincadl itcihoenmails trwy inatn ethre. University of Tübingen. (Source: Universitäts- archiv TübingeWn, eC ahtalvoeg uael lN boe. e1n26 r/a7i6s)ed in the conceptions of Pasteur. When experimental facts came into my hands that in thDe ubriionlgo gai cvails ict otuor shei so fo ledveern tbsr, owthheicrh H waonus ldin nMevuenr icbhe icno mOpclteotbeelyr appeared to argue in favour of cell-free fermentation, I u1n89d6e,r sEtodouda…rd TBhuicsh hnyepro thhaeds ism oafn tahgee