FEMS Yeast Research Advance Access published June 24, 2015
In Delft: a personal account ABSTRACT The author looks back on his development in microbiology and yeast research, and on the establishement in Delft of the FEMS Central Office, FEMS Publications Office, and the birth of FEMS Yeast Research. KEYWORDS History, Delft, Kluyver Laboratory, Yeast research AUTHORS’S ADDRESS W. Alexander Scheffers c/o Department of Microbiology Delft University of Technology Julianalaan 67A NL-2628 BC The Netherlands [email protected] In Delft, Antoni van Leeuwenhoek, upon observing in 1674, with his home-made little lens, for the first time bacteria, described in 1680 the presence of yeast in beer. However, apart from the many breweries along the canals, it then took centuries before yeasts attained an outstanding place in Delft. In 1870, in this town J.C. van Marken founded the Gistfabriek (yeast factory) for the production of baker’s yeast and alcohol; there, in 1885, Martinus Willem Beijerinck became the head of a new microbiological laboratory. Thereupon, being appointed in 1895 as professor of microbiology at the then “Polytechnical School” in the same town, he demanded a laboratory annex private house to be built at the Nieuwelaan along the big canal (Fig. 1). Many of his famous enrichment cultures for a great variety of bacteria have been started with inocula from the canal or from his large garden. His successor in 1921, Albert Jan Kluyver (Ph.D. Thesis Delft , 1914; Kamp et al., 1959) made an audacious new start. With his co-worker he developed the revolutionary vision of “the unity in biochemistry” (Kluyver & Donker, 1926). From the graduates who in the course of years took their Ph.D. under Kluyver’s direction, a considerable part worked on yeasts: Struyk (1928), Stelling-Dekker (1931), Lodder (1934), Hoogerheide (1935), Custers (1940) and van der Walt (1952). Besides, Bulder (1963) formally obtained his degree under the successor. A remarkable achievement are the pioneering micrographs of bacteria and yeasts with the Delft prototype electron microscope (Houwink et al., 1949). As a student in chemical engineering, I became fascinated during the laboratory course on microbes, given personally by Kluyver (Fig. 2), with his assistant Verhoeven, during evening hours, and later by his lectures in the small theatre at the Nieuwelaan. I had found my destination! I decided to do the final stage of my study at Kluyver's laboratory. The subject was bacterial slime formation in a paper mill. Kluyver conducted me to the factory where we took samples for analysis in Delft. When, unexpectedly for us, Kluyver died in 1956, the laboratory was in grief. We had lost our beloved master (Fig. 3). In 1957 the successor, the Swede Torsten Wikén, arrived. In earlier years he had made a short traineeship with Kluyver and further for ten years had filled a professorship in microbiology at the ETH in Zürich. With him I formally had to make my final examination. When I expressed the wish to continue in
microbiology, he had no means to appoint me. Thereupon I independently succeeded in obtaining a small grant from the Delft University Fund. I was intrigued by the "negative Pasteur effect", described by Custers (1940) in Brettanomyces: the inhibition of fermentation under anaerobic conditions, and decided to continue these studies . At the Nieuwelaan, I developed my techniques for manometric measurement of yeast metabolism under aerobic and strictly anaerobic conditions (Fig. 4).
The next year, in 1958, we moved to the new institute at Julianalaan (Fig. 5). The laboratory, connected to the existing institutes of biochemistry and technical botany, had been designed by Kluyver, inclusive the professor's residence at the side. Alas, he had not lived to see its completion. Meanwhile I made good progress with the manometric Warburg experiments. In 1960, Wikén could appoint me as scientific assistant (Fig. 6). My experiments led me to the insight that the negative Pasteur effect resulted from a disturbance in the redox balance of the NAD+/NADH couple (Scheffers, 1961, 1966, 1967; Tromp & Scheffers, 1969; Scheffers & Misset, 1974). Over the years, various other studies were carried out on yeasts (Wikén et al.,1961, 1962; Tromp et al. 1968; Scheffers & Wikén, 1969; Smith et al., 1981; van Doorne et al., 1984), as well as on food microbiology (Bakker et al., 1968; González et al, 1971; Dijkstra et al., 1972; Golten et al., 1975; de Boer et al., 1975 a,b; Havelaar et al., 1980). Moreover, I conducted the scientific organization of the International Symposium on Analytical Methods and Problems in Biotechnology (Scheffers et al., 1984); a sequel followed four years later (te Nijenhuis et al. 1988). A favourable circumstance was the cohabitation, since Kluyver’s time, of the CBS culture collection of yeasts and our institute. Particularly Nel Kreger-van Rij (Fig. 7) guided me in the systematics of yeasts (Lodder & Kreger-van Rij, 1952, 1964) and provided many cultures from diverging genera for comparative manometric studies (Scheffers & Wikén, 1969). Meanwhile I had been commissioned with a variety of tasks in the department: administration of personnel affairs, management of the chemicals store, developing practical and lecture courses in general, sanitary, and food microbiology, coaching students in their individual research tasks. With a colleague, Peter Arntz, we monitored the microbiological quality of drinking and swimming water around the town, and with the university’s student doctor I monitored the various student canteens and kitchens for cleanliness and pathogens. With all that my own research was the closing entry. Gradually I learned to go my own way (Pernice et al., 1970; Golten & Scheffers, 1975; de Boer et al., 1975 a,b; Alonzo & Scheffers, 1975; Havelaar et al., 1980; Carrascosa et al., 1981; Smith et al., 1981; Nicolay et al. 1982, 1983). Summer 1977, Wikén unexpectedly left in retirement to his native country Sweden, leaving behind a department with meanwhile nine staff members, with administrative and laboratory personnel, and with students of various grades. The faculty then appointed me as the department’s acting chairman, a function that I fulfilled till 1980. First of all, during my summer holidays in 1977, I rushed into the preparation of my lecture course in general microbiology, to be started in September and to be presented in a new, contemporary form and content. I also organized, in the framework of the North-West European Microbiology Group’s meeting of 1977 in Amsterdam, a symposium on marine microbiology, with Rita Colwell as the main speaker. A new period started in 1980 with the appointment of Gijs Kuenen (Fig. 8) as head of the department. With him from Groningen came Hans van Dijken, who joined me in a new start of the yeast physiology project. This was the beginning of a fruitful co-operation, resulting in a flow of papers over the years to come. Besides, from 1980 till 1984, I served as the general secretary of the recently founded Netherlands Biotechnological Society. In continuation on my earlier work (Scheffers, 1961, 1966) we elaborated the role in yeasts of the redox couples NAD+/NADH and NADP+/NADPH (Bruinenberg et al., 1983 a,b; 1985 a,b). Subsequently it turned
out that these coenzymes play a determining role in the fermentation of xylose (Toivola et al., 1984; Bruinenberg et al., 1983, 1984 a,b; Verduyn et al.,1985 a,b,c; van Dijken & Scheffers, patent 1985; Pronk et al., 1988). Further work on these redox couples resulted in papers by Wijsman et al. (1984), Bruinenberg et al. (1985 a,b), and a review by van Dijken & Scheffers (1986). The isolation of mitochondria and exploration of their role in yeast metabolism were tackled in a series of articles (Bruinenberg et al., 1985 a,b; van Urk et al., 1989; Verduyn et al., 1991). In the meantime other, related work was published on fermentation in “non-fermentative” yeasts (van Dijken et al., 1986), radiorespirometric study of glucose metabolism (Bruinenberg et al., 1986 a,b), orthogonal-field-alternation banding patterns of DNA from yeasts (de Jonge et al., 1986), adenylate cyclase of yeast (Purwin et al., 1986; Noshiro et al., 1987). Besides, analytical methods were developed for measurement of alcohol production (Verduyn et al., 1983, 1984 a,b), and for protein determination (Rouwenhorst et al., 1991). A variety of enzymological aspects of yeasts came to the fore in articles on alcohol dehydrogenases (Verduyn et al., 1988 a), metabolism of 2,3-butanediol and dihydroxyacetone (Verduyn et al., 1988 b,c), hydrogen peroxide (Verduyn et al., 1988 d), pyruvate (van Urk et al., 1989), inulin (Rouwenhorst et al., 1988, 1990 a,b, 1991; Hensing et al., 1993). Sugar transport in yeasts led us to a series of papers: van Urk et al. (1989); Postma et al. (1988,1989, 1990), Weusthuis et al. (1993). At the same time, also many papers dealing with aspects of regulation appeared: van Urk et al. (1988, 1990); Postma et al. (1989 a,b); Verduyn et al. (1990 a,b, 1991, 1992), Visser et al. (1990); Weusthuis et al. (1994 a,b); Pronk et al. (1994 a,b). Besides, I got involved in studies on the molecular biology of yeasts (Boekhout et al., 1993, 1997). From 1987 till 1997 I was a member of the CBS Science Commission. In 1990, my official, obligatory retirement from the university procured me time to take up new tasks. I became a board member of the Netherlands Society for Microbiology and founded its Section Mycology (Scheffers, 1994). As members of the International Commission on Yeasts, Hans van Dijken and I organized the 16th International Specialized Symposium on Yeasts on Metabolic Compartmentation (Scheffers & van Dijken, 1993). In 1995 we took part in the organization of the Beijerinck Centennial (Scheffers & van Dijken, 1995), in 1995 I contributed a commemorative paper on the centennial of the Delft School of Microbiology (Scheffers, 1996) and in 1999 I edited the contributed papers of the Eijkman Centennial (Scheffers, 1999). In 1997, the FEMS Executive Board asked me to investigate the possibility of establishing, in The Netherlands, a Central Office for its organization. In view of the historical part of Delft in the development of microbiology, I saw this as the right place. So I was happy to obtain permission for the use of a few rooms in the former professor’s house adjacent to the laboratory of microbiology of the Delft University of Technology. In this office the FEMS Board Members assembled shortly after that, to see the applicants. Diman van Rossum was then selected as the first Executive Officer of FEMS Central Office (1998-2006). He was supported by a small administrative staff. Soon followed the appointment of Wilma van Wezenbeek as the Executive Officer for FEMS Publications (2000-2006) and her assistant Gillian van Beest. Shortly afterwards, FEMS Executive Board invited me to set up its fifth journal, FEMS Yeast Research. An important task would be to find eligible candidates for the future editorial board. An excellent opportunity presented itself during the Tenth International Symposium on Yeasts, organized in 2000 at Papendal (Arnhem) (van Dijken & Scheffers, 2000). The invited candidates showed great approval and soon I could start with an outstanding editorial board. The new journal developed favourably, and after five years I
could turn over chief editorship to Teun Boekhout for another five years. Since then the task is in the able hands of Jens Nielsen. In 2003 I was honoured with the FEMS Special Award (Fig. 9). I enjoyed being involved in so many aspects of microbiology, in particular of yeasts. The concerted action with so many capable colleagues and friends was a great privilege and joy. I was happy to be a co-author in a collective paper (van Maris et al., 2006) by the group of Jack Pronk (Fig. 10). And I was delighted when our xylose-fermenting yeast, Pichia stipitis, was relegated to the new genus Scheffersomyces (Kurtzman & Suzuki, 2010). I congratulate FEMS at the occasion of its 40th anniversary and wish it a bright and golden future ! And may FEMS Yeast Research continue to be a valuable and esteemed offspring on its stock!
ACKNOWLEDGEMENT To Cocky, my wife, and our children Peter and Irene, for their love and support throughout the years.
REFERENCES Alonzo V & Scheffers WA (1975) Studies on hydrolysis and synthesis of fats in Staphylococcus aureus and Candida lipolytica. Boll Ist Sieroter Milanese 54: 90-97. Bakker G, Scheffers WA & Wikén TO (1968) A new method for the determination of clotting times in milk. Neth Milk and Dairy J 22: 16-21. Boekhout T, Renting M, Scheffers WA & Bosboom R (1993) The use of karyotyping in the systematics of yeasts. Antonie van Leeuwenhoek 63: 157-163. Boekhout T, van Belkum A, Leenders ACAP, Verbrugh HA, Mukamurangwa P, Swinne D & Scheffers WA (1997) Molecular typing of Cryptococcus neoformans: taxonomic and epidemiological aspects. Internatl. J Syst Bacteriol 47: 432-442. Bruinenberg PM, de Bot PHM, van Dijken JP & Scheffers WA (1983) The role of redox balances in the anaerobic fermentation of xylose by yeasts. Eur J Appl Microbiol Biotechnol 18: 287-292. Bruinenberg PM, de Bot PHM, van Dijken JP & Scheffers WA (1984) NADH-linked aldose reductase: the key to anaerobic alcoholic fermentation of xylose by yeasts. Appl Microbiol Biotechnol 19: 256-260. Bruinenberg PM, Jonker R, van Dijken JP & Scheffers WA (1985) Utilization of formate as an additional energy source by glucose-limited chemostat cultures of Candida utilis CBSB21 and Saccharomyces cerevisiae CBS8066. Evidence for the absence of transhydrogenase activity in yeasts. Arch Microbiol 142: 302-306. Bruinenberg PM, van Dijken JP, Kuenen JG & Scheffers WA(1985) Critical parameters in the isolation of mitochondria from Candida utilis grown in continuous culture. J Gen Microbiol 131: 1035-1042. Bruinenberg PM, van Dijken JP, Kuenen JG & Scheffers WA (1985) Oxidation of NADH and NADPH by mitochondria from the yeast Candida utilis. J Gen Microbiol 131: 1043-1051. Bruinenberg PM, van Dijken JP & Scheffers WA (1983) A theoretical analysis of NADPH production and consumption in yeasts. J Gen Microbiol 129: 953-964. Bruinenberg PM, van Dijken JP & Scheffers WA (1983) An enzymic analysis of NADPH production and consumption in Candida utilis. J Gen Microbiol 129: 965-971. Bruinenberg PM, van Dijken JP & Scheffers WA (1986) A radiorespirometric study on the contribution of the hexose monophosphate pathway to glucose metabolism in Candida utilisCB3621 grown in glucose-limited chemostat cultures. J Gen Microbiol 132: 221-229. Bruinenberg PM, Waslander GW, van Dijken JP & Scheffers WA (1986) A comparative radiorespirometric study of glucose metabolism in yeasts. Yeast 2: 117-121. Bulder CJEA. On Respiratory Deficiency in Yeasts. Ph.D. Thesis, Delft. Groningen: VRB, 1963.
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glucose between the yeasts Saccharomyces cerevisiae and Candida utilis. Appl Environ Microbiol 55: 3214-3220. Postma E, Scheffers WA & van Dijken JP (1988) Adaptation of the kinetics of glucose transport to environmental conditions in the yeast Candida utilis CBS621: a continuous-culture study. J gen Microbiol 134:1109-1116. Postma E, Scheffers WA & van Dijken JP (1989) Kinetics of growth and glucose transport in glucose-limited chemostat cultures of Saccharomyces cerevisiae CBS8066. Yeast 5:159-165. Postma E, Verduyn C, Kuiper A, Scheffers WA & van Dijken JP (1990) Substrate-accelerated death of Saccharomyces cerevisiae CBS8066 under maltose stress. Yeast 6:149-158. Postma E, Verduyn C, Scheffers WA & van Dijken JP (1989) Enzymic analysis of the Crabtree effect in glucose-limited chemostat cultures of Saccharomyces cerevisiae. Appl Environ Microbiol 55: 468-477. Pronk JT, Bakker AW, van Dam HE, Straathof AJJ, Scheffers WA & van Dijken JP (1988) Preparation of D-xylulose from D-xylose. Enzyme Microb Technol 10:537-542. Pronk JT, van der Linden-Beuman A, Verduyn C, Scheffers WA & van Dijken JP (1994) Propionate metabolism in Saccharomyces cerevisiae: implications for the metabolon hypothesis. Microbiology 140: 717-722. Pronk JT, Wenzel TJ, Luttik MAH, Klaassen CCM, Scheffers WA, Steensma HY & van Dijken JP (1994) Energetic aspects of glucose metabolism in a pyruvate-dehydrogenase-deficient mutant of Saccharomyces cerevisiae. Microbiology 140: 601-610. Purwin C, Nicolay K, Scheffers WA & Holzer H (1986) Mechanism of control of adenylate cyclase activity in yeast by fermentable sugars and carbonyl cyanide m-chlorophenyl hydrazone. J Biol Chem 261: 8744-8749. Rouwenhorst RJ, Frank J, Scheffers WA & van Dijken JP (1991) Determination of protein concentration by total carbon analysis (1991) J Biophys Methods 22:119-128. Rouwenhorst RJ, Hensing M, Verbakel J, Scheffers WA & van Dijken JP (1990) Structure and properties of the extracellular inulinase of Kluyveromyces marxianus CBS6556. Appl Environ Microbiol 56:3337-3345. Rouwenhorst RJ, Ritmeester WS, Scheffers WA & van Dijken JP (1990) Localization of inulinase and invertase in Kluyveromyces species. Appl Environ Microbiol 56: 3329-3336. Rouwenhorst RJ, van der Baan AA, Scheffers WA & van Dijken JP (1991) Production and localization of B-fructosidase in asynchronous and synchronous cultures of yeasts. Appl Environ Microbiol 57:557-562. Rouwenhorst RJ, Visser LE, van der Baan AA, Scheffers WA & van Dijken JP (1988) Production, distribution, and kinetic properties of inulinase in continuous cultures of Kluyveromyces marxianus CBS 6556.Appl Environ Microbiol 54: 1131-1137. Scheffers WA (1961) On the inhibition of alcoholic fermentation in Brettanomyces yeasts under anaerobic conditions. Experientia 17:40-42. Scheffers WA (1966) Stimulation of fermentation in yeasts by acetoin and oxygen. Nature 210: 533534. Scheffers WA (1967) Effects of oxygen and acetoin on fermentation and growth in Brettanomyces and some other yeast genera. Atti del XIV Congr Soc Ital Microbiol: 91-107. Scheffers WA (1994) The Netherlands Society for Microbiology/ Nederlandse Vereniging voor Microbiologie (NvvM). SGM Quarterly: 77-78. Scheffers WA . Honderd jaar leerstoel microbiologie Delft, 1895-1995. In: Jaarboek Nederlandse Vereniging voor Microbiologie 1996, 11-15. Scheffers WA (1999) Eijkman Centennial on Infections in the 21st Century. “Successes from the past, challenges for the future”. FEMS Immunol Medic Microbiol 26: 183-321. Scheffers WA & Misset O (1974) The Custers effect in Brettanomyces intermedius. Proc. 4 th Int Symp on Yeasts, Vienna, Part I: 39-40. Scheffers WA, te Nijenhuis B & Kragten J (eds) (1984) Analytical Methods and Problems in Biotechnology. Proceed Internatl Symp, Noordwijkerhout. Analytica Chim Acta 163: 1-328.
Scheffers WA & van Dijken JP (eds). Metabolic Compartmentation in Yeasts. 16th Internatl Specialized Symp on Yeasts (ISSY 16, Arnhem,1993). The Hague: Pasmans, 1993. Scheffers WA & van Dijken JP (eds). Beijerinck Centennial. Microbial Physiology and Gene Regulation: Emerging Principles and Applications. Delft: Delft Univ. Press, 1995. Scheffers WA & Wikén TO (1969) The Custers effect (negative Pasteur effect) as a taxonomic criterion for the genus Brettanomyces. Antonie van Leeuwenhoek 35: A31-A32. Smith MT, Batenburg-van der Vegte WH & Scheffers WA (1981) Eeniella, a new yeast genus of the Torulopsidales. Internatl J Syst Bacteriol 31:196-203. Stelling-Dekker NM. Beiträge zu einer Monographie der Hefearten. I. Teil. Die Sporogenen Hefen. PhD Thesis, Utrecht, 1931. Struyk AP. Onderzoekingen over de Alcoholische Gisting. PhD Thesis, Delft, 1928. te Nijenhuis B, van Dijk C, Kragten J & Scheffers WA (eds) (1988) Analytical Methods and Problems in Biotechnology. Proceed 2 nd Internatl Symp, Anabiotec'88, Noordwijkerhout. Analytica Chim Acta 213:1-183. Toivola A, Yarrow D, van den Bosch E, van Dijken JP & Scheffers WA (1984) Alcoholic fermentation of D-xylose by yeasts. Appl Environ Microbiol 47:1221-1223. Tromp AGGM, Bonnet JABAF & Scheffers WA (1968) A simple culture vessel for turbidimetric studies of anaerobic growth of microbes. Experientia 24:1076-1077. Tromp AGGM & Scheffers WA (1969) Effect of acetoin on anaerobic growth in Brettanomyces. Antonie van Leeuwenhoek 35: H37-H38. van der Walt JP. On the Yeast Candida pulcherrima and its Pigment. PhD Thesis, Delft. van Dijken JP & Scheffers WA. Studies on alcoholic fermentation in yeasts. In: Houwink EH & van der Meer (eds). Innovations in Biotechnology. Amsterdam: Elsevier Science Publ, 1984. van Dijken JP & Scheffers WA (1985) Procedé de production d' éthanol à partir d' une substance contenant du xylose. Patent: FR 2545838 (A1). van Dijken JP & Scheffers WA (1986) Redox balances in the metabolism of sugars by yeasts. FEMS Microbiol Reviews 32: 199-224. van Dijken JP & Scheffers WA (eds) The Rising Power of Yeasts in Science and Industry. ISY 2000. Tenth Internatl. Symp. On Yeasts, Arnhem, The Netherlands. Delft University Press, 2000. van Dijken JP, van den Bosch E, Hermans JJ, Rodrigues de Miranda L & Scheffers WA (1986) Alcoholic fermentation by “non-fermentative”yeasts. Yeast 2: 123-127. van Doorne H, Scheffers WA, Hadiutomo M & van den Bosch E (1984) Microbial contamination of a vitamin A formulation, prepared in local pharmacies, and its preservation against yeasts and moulds. Antonie van Leeuwenhoek 50: 405-416. van Maris AJA, Abbott DA, Bellissimi E, van den Brink J, Kuyper M, Luttik MAH, Wisselink HW, Scheffers WA, van Dijken JP & Pronk JT (2006) Alcoholic fermentation of carbon sources in biomass hydrolysates by Saccharomyces cerevisiae: current status. Antonie van Leeuwenhoek 90: 391-418. van Urk H, Bruinenberg PM, Veenhuis M, Scheffers WA & van Dijken JP (1989) Respiratory capacities of mitochondria of Saccharomyces cerevisae CBS 8066 and Candida utilis CBS 621 grown under glucose limitation. Antonie van Leeuwenhoek 56: 211-220. van Urk H, Mak PR, Scheffers WA & van Dijken JP (1988) Metabolic responses of iSaccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621upon transition from glucose limitation to glucose excess. Yeast 4: 283-291. van Urk H, Postma E, Scheffers WA & van Dijken JP (1989) Glucose transport in Crabtree-positive and Crabtree-negative yeasts. J Gen Microbiol 135: 2399-2406. van Urk, H, Schipper D, Breedveld GJ, Mak PR, Scheffers WA & van Dijken JP (1989) Localization and kinetics of pyruvate-metabolizing enzymes in relation to aerobic alcoholic fermentation in Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621. Biochim Biophys Acta 992: 78-86. van Urk H, Voll WSL, Scheffers WA & van Dijken JP (1990) Transient-state analysis of metabolic fluxes in Crabtree-positive and Crabtree-negative yeasts. Appl Environ Microbiol.56:281-287.
Verduyn C, Breedveld GJ, Scheffers WA & van Dijken JP (1988) Purification and properties of dihydroxyacetone reductase and 2,3-butanediol dehydrogenase from Candida utilis CBS 621. Yeast 4: 127-133. Verduyn C, Breedveld GJ, Scheffers WA & van Dijken JP (1988) Metabolism of 2-3-butanediol in yeasts. Yeast 4: 135-142. Verduyn C, Breedveld GJ, Scheffers WA & van Dijken JP (1988) Substrate specificity of alcohol dehydrogenase from the yeasts Hansenula polymorpha CBS 4732 and Candida utilis CBS 621. Yeast 4:143-148. Verduyn C, Breedveld GJ, Schreuder H, Scheffers WA & van Dijken JP (1988) Properties of enzymes which reduce dihydroxyacetone and related compounds in Hansenula polymorpha CBS 4732. Yeast 4: 117-126. Verduyn C, Frank J, van Dijken JP & Scheffers WA (1985) Multiple forms of xylose reductase in Pachysolen tannophilus. FEMS Microbiol Lett 30: 313-317. Verduyn C, Giuseppin MLF, Scheffers WA & van Dijken JP (1988) Hydrogen peroxide metabolism in yeasts. Appl. Environ. Microbiol. 54: 2086-2090. Verduyn C, Postma E., Scheffers WA & van Dijken JP (1990) Physiology of Saccharomyces cerevisiae in anaerobic glucose-limited chemostat cultures. J Gen Microbiol 136: 395-403. Verduyn C, Postma E, Scheffers WA & van Dijken JP (1990) Energetics of Saccharomyces cerevisiae in anaerobic glucose-limited chemostat cultures. J Gen Microbiol 136: 405-412. Verduyn C, Postma E, Scheffers WA & van Dijken JP (1992) Effect of benzoic acid on metabolic fluxes in yeasts: a continuous-culture study on the regulation of respiration and alcoholic fermentation. Yeast 8: 501-517. Verduyn C, Stouthamer AH, Scheffers WA & van Dijken JP (1991) A theoretical evaluation of growth yields of yeasts. Antonie van Leeuwenhoek 59: 49-63. Verduyn C, van Dijken JP & Scheffers WA (1983) A simple, sensitive, and accurate alcohol electrode. Biotechnol Bioeng 25: 1049-1055. Verduyn C, van Dijken JP & Scheffers WA (1984) Colorimetric alcohol assays with alcohol oxidase. J Microbiol Methods 2: 15-25. Verduyn C, van Kleef R, Frank J, Schreuder H, van Dijken JP & Scheffers WA (1985) Properties of the NAD(P)H-dependent xylose reductase from the xylose-fermenting yeast Pichia stipitis. Biochem J 226: 669-677. Verduyn C, van Wijngaarden CJ, Scheffers WA & van Dijken JP (1991) Hydrogen peroxide as an electron acceptor for mitochondrial respiration in the yeast Hansenula polymorpha. Yeast 7: 137146. Verduyn C, Zomerdijk TPL, van Dijken JP & Scheffers WA (1984) Continuous measurement of ethanol production by aerobic yeast suspensions with an enzyme electrode. ApplMicrobiol Biotechnol 19: 181-185. Visser W, Scheffers WA, Batenburg-van der Vegte WH & van Dijken JP (1990) Oxygen requirements of yeasts. Appl Environ Microbiol 56: 3785-3792. Visser W, van der Baan AA, Batenburg-van der Vegte WH, Scheffers WA, Krämer R & van Dijken JP (1994) Involvement of mitochondria in the assimilatory metabolism of anaerobic Saccharomyces cerevisiae cultures. Microbiology 140: 3039-3046. Weusthuis RA, Adams H, Scheffers WA & van Dijken JP (1993) Energetics and kinetics of maltose transport in Saccharomyces cerevisiae: a continuous-culture study. Appl Environ Microbiol 59: 3102-3109. Weusthuis RA, LuttikMAH, Scheffers WA, van Dijken JP & Pronk JP (1994) Is the Kluyver effect in yeasts caused by product inhibition? Microbiology 140: 1723-1729. Weusthuis RA, Visser W, Pronk JP, Scheffers WA & van Dijken JP (1994) Effects of oxygen limitation on sugar metabolism in yeasts: a continuous-culture study of the Kluyver effect. Microbiology 140:703-715. Wijsman MR, van Dijken JP, van Kleeff BHA& Scheffers WA (1984) Inhibition of fermentation and growth in batch cultures of the yeast Brettanomyces intermedius upon a shift from aerobic to
anaerobic conditions (Custers effect). Antonie van Leeuwenhoek 50: 183-192. Wikén T, Scheffers WA & Verhaar AJM (1961) On the existence of a negative Pasteur effect in yeasts classified in the genus Brettanomyces Kufferath et van Laer. Antonie van Leeuwenhoek 27: 401-433. Wiken T, Verhaar AJM & Scheffers WA (1962) The influence of potassium and sodium ions on the negetive Pasteur effect in Brettanomyces claussenii Custers. Archiv f. Mikrobiol. 42: 226-236.
FIGURE LEGENDS
Fig.1 The Professor’s residence (left part) connected to the laboratory (right) of Beijerinck and Kluyver at Nieuwelaan 3
Fig.2 Kluyver in the library. Photo by J.A. Schuur
Fig.3 Kluyver in his workroom with van Niel, during the preparation of their book (1955). Photo by J.K. Baars
Fig. 4 The author at a manometric experiment, measuring aerobic and anaerobic fermentation and respiration rates of yeasts (1958)
Fig. 5 The laboratory of Julianalaan
Fig. 6 Wikén with the author. Photo by J.A. Schuur
Fig. 7 Nel Kreger-van Rij, Yeast Division of Centraalbureauu voor Schimmelcultures (CBS)
Fig. 8 Gijs Kuenen with Peter Arntz, at the latter’s silver jubilee (1980). Photo by J.A. Schuur
Fig. 9 FEMS Special Award, presented to the author during the first FEMS Symposium, Ljubljana (2003). Photo by Diman van Rossum
Fig. 10 The author, Jack Pronk and Hans van Dijken (from left to right) on the lake of Bled, Slovenia, during the International Specialised Symposium on Yeasts (1997). Photo by Leonie Raamsdonk
In Delft: a personal account ABSTRACT The author looks back on his development in microbiology and yeast research, and on the establishement in Delft of the FEMS Central Office, FEMS Publications Office, and the birth of FEMS Yeast Research. KEYWORDS History, Delft, Kluyver Laboratory, Yeast research AUTHORS’S ADDRESS W. Alexander Scheffers c/o Department of Microbiology Delft University of Technology Julianalaan 67A NL-2628 BC The Netherlands [email protected] In Delft, Antoni van Leeuwenhoek, upon observing in 1674, with his home-made little lens, for the first time bacteria, described in 1680 the presence of yeast in beer. However, apart from the many breweries along the canals, it then took centuries before yeasts attained an outstanding place in Delft. In 1870, in this town J.C. van Marken founded the Gistfabriek (yeast factory) for the production of baker’s yeast and alcohol; there, in 1885, Martinus Willem Beijerinck became the head of a new microbiological laboratory. Thereupon, being appointed in 1895 as professor of microbiology at the then “Polytechnical School” in the same town, he demanded a laboratory annex private house to be built at the Nieuwelaan along the big canal (Fig. 1). Many of his famous enrichment cultures for a great variety of bacteria have been started with inocula from the canal or from his large garden. His successor in 1921, Albert Jan Kluyver (Ph.D. Thesis Delft , 1914; Kamp et al., 1959) made an audacious new start. With his co-worker he developed the revolutionary vision of “the unity in biochemistry” (Kluyver & Donker, 1926). From the graduates who in the course of years took their Ph.D. under Kluyver’s direction, a considerable part worked on yeasts: Struyk (1928), Stelling-Dekker (1931), Lodder (1934), Hoogerheide (1935), Custers (1940) and van der Walt (1952). Besides, Bulder (1963) formally obtained his degree under the successor. A remarkable achievement are the pioneering micrographs of bacteria and yeasts with the Delft prototype electron microscope (Houwink et al., 1949). As a student in chemical engineering, I became fascinated during the laboratory course on microbes, given personally by Kluyver (Fig. 2), with his assistant Verhoeven, during evening hours, and later by his lectures in the small theatre at the Nieuwelaan. I had found my destination! I decided to do the final stage of my study at Kluyver's laboratory. The subject was bacterial slime formation in a paper mill. Kluyver conducted me to the factory where we took samples for analysis in Delft. When, unexpectedly for us, Kluyver died in 1956, the laboratory was in grief. We had lost our beloved master (Fig. 3). In 1957 the successor, the Swede Torsten Wikén, arrived. In earlier years he had made a short traineeship with Kluyver and further for ten years had filled a professorship in microbiology at the ETH in Zürich. With him I formally had to make my final examination. When I expressed the wish to continue in
microbiology, he had no means to appoint me. Thereupon I independently succeeded in obtaining a small grant from the Delft University Fund. I was intrigued by the "negative Pasteur effect", described by Custers (1940) in Brettanomyces: the inhibition of fermentation under anaerobic conditions, and decided to continue these studies . At the Nieuwelaan, I developed my techniques for manometric measurement of yeast metabolism under aerobic and strictly anaerobic conditions (Fig. 4).
The next year, in 1958, we moved to the new institute at Julianalaan (Fig. 5). The laboratory, connected to the existing institutes of biochemistry and technical botany, had been designed by Kluyver, inclusive the professor's residence at the side. Alas, he had not lived to see its completion. Meanwhile I made good progress with the manometric Warburg experiments. In 1960, Wikén could appoint me as scientific assistant (Fig. 6). My experiments led me to the insight that the negative Pasteur effect resulted from a disturbance in the redox balance of the NAD+/NADH couple (Scheffers, 1961, 1966, 1967; Tromp & Scheffers, 1969; Scheffers & Misset, 1974). Over the years, various other studies were carried out on yeasts (Wikén et al.,1961, 1962; Tromp et al. 1968; Scheffers & Wikén, 1969; Smith et al., 1981; van Doorne et al., 1984), as well as on food microbiology (Bakker et al., 1968; González et al, 1971; Dijkstra et al., 1972; Golten et al., 1975; de Boer et al., 1975 a,b; Havelaar et al., 1980). Moreover, I conducted the scientific organization of the International Symposium on Analytical Methods and Problems in Biotechnology (Scheffers et al., 1984); a sequel followed four years later (te Nijenhuis et al. 1988). A favourable circumstance was the cohabitation, since Kluyver’s time, of the CBS culture collection of yeasts and our institute. Particularly Nel Kreger-van Rij (Fig. 7) guided me in the systematics of yeasts (Lodder & Kreger-van Rij, 1952, 1964) and provided many cultures from diverging genera for comparative manometric studies (Scheffers & Wikén, 1969). Meanwhile I had been commissioned with a variety of tasks in the department: administration of personnel affairs, management of the chemicals store, developing practical and lecture courses in general, sanitary, and food microbiology, coaching students in their individual research tasks. With a colleague, Peter Arntz, we monitored the microbiological quality of drinking and swimming water around the town, and with the university’s student doctor I monitored the various student canteens and kitchens for cleanliness and pathogens. With all that my own research was the closing entry. Gradually I learned to go my own way (Pernice et al., 1970; Golten & Scheffers, 1975; de Boer et al., 1975 a,b; Alonzo & Scheffers, 1975; Havelaar et al., 1980; Carrascosa et al., 1981; Smith et al., 1981; Nicolay et al. 1982, 1983). Summer 1977, Wikén unexpectedly left in retirement to his native country Sweden, leaving behind a department with meanwhile nine staff members, with administrative and laboratory personnel, and with students of various grades. The faculty then appointed me as the department’s acting chairman, a function that I fulfilled till 1980. First of all, during my summer holidays in 1977, I rushed into the preparation of my lecture course in general microbiology, to be started in September and to be presented in a new, contemporary form and content. I also organized, in the framework of the North-West European Microbiology Group’s meeting of 1977 in Amsterdam, a symposium on marine microbiology, with Rita Colwell as the main speaker. A new period started in 1980 with the appointment of Gijs Kuenen (Fig. 8) as head of the department. With him from Groningen came Hans van Dijken, who joined me in a new start of the yeast physiology project. This was the beginning of a fruitful co-operation, resulting in a flow of papers over the years to come. Besides, from 1980 till 1984, I served as the general secretary of the recently founded Netherlands Biotechnological Society. In continuation on my earlier work (Scheffers, 1961, 1966) we elaborated the role in yeasts of the redox couples NAD+/NADH and NADP+/NADPH (Bruinenberg et al., 1983 a,b; 1985 a,b). Subsequently it turned
out that these coenzymes play a determining role in the fermentation of xylose (Toivola et al., 1984; Bruinenberg et al., 1983, 1984 a,b; Verduyn et al.,1985 a,b,c; van Dijken & Scheffers, patent 1985; Pronk et al., 1988). Further work on these redox couples resulted in papers by Wijsman et al. (1984), Bruinenberg et al. (1985 a,b), and a review by van Dijken & Scheffers (1986). The isolation of mitochondria and exploration of their role in yeast metabolism were tackled in a series of articles (Bruinenberg et al., 1985 a,b; van Urk et al., 1989; Verduyn et al., 1991). In the meantime other, related work was published on fermentation in “non-fermentative” yeasts (van Dijken et al., 1986), radiorespirometric study of glucose metabolism (Bruinenberg et al., 1986 a,b), orthogonal-field-alternation banding patterns of DNA from yeasts (de Jonge et al., 1986), adenylate cyclase of yeast (Purwin et al., 1986; Noshiro et al., 1987). Besides, analytical methods were developed for measurement of alcohol production (Verduyn et al., 1983, 1984 a,b), and for protein determination (Rouwenhorst et al., 1991). A variety of enzymological aspects of yeasts came to the fore in articles on alcohol dehydrogenases (Verduyn et al., 1988 a), metabolism of 2,3-butanediol and dihydroxyacetone (Verduyn et al., 1988 b,c), hydrogen peroxide (Verduyn et al., 1988 d), pyruvate (van Urk et al., 1989), inulin (Rouwenhorst et al., 1988, 1990 a,b, 1991; Hensing et al., 1993). Sugar transport in yeasts led us to a series of papers: van Urk et al. (1989); Postma et al. (1988,1989, 1990), Weusthuis et al. (1993). At the same time, also many papers dealing with aspects of regulation appeared: van Urk et al. (1988, 1990); Postma et al. (1989 a,b); Verduyn et al. (1990 a,b, 1991, 1992), Visser et al. (1990); Weusthuis et al. (1994 a,b); Pronk et al. (1994 a,b). Besides, I got involved in studies on the molecular biology of yeasts (Boekhout et al., 1993, 1997). From 1987 till 1997 I was a member of the CBS Science Commission. In 1990, my official, obligatory retirement from the university procured me time to take up new tasks. I became a board member of the Netherlands Society for Microbiology and founded its Section Mycology (Scheffers, 1994). As members of the International Commission on Yeasts, Hans van Dijken and I organized the 16th International Specialized Symposium on Yeasts on Metabolic Compartmentation (Scheffers & van Dijken, 1993). In 1995 we took part in the organization of the Beijerinck Centennial (Scheffers & van Dijken, 1995), in 1995 I contributed a commemorative paper on the centennial of the Delft School of Microbiology (Scheffers, 1996) and in 1999 I edited the contributed papers of the Eijkman Centennial (Scheffers, 1999). In 1997, the FEMS Executive Board asked me to investigate the possibility of establishing, in The Netherlands, a Central Office for its organization. In view of the historical part of Delft in the development of microbiology, I saw this as the right place. So I was happy to obtain permission for the use of a few rooms in the former professor’s house adjacent to the laboratory of microbiology of the Delft University of Technology. In this office the FEMS Board Members assembled shortly after that, to see the applicants. Diman van Rossum was then selected as the first Executive Officer of FEMS Central Office (1998-2006). He was supported by a small administrative staff. Soon followed the appointment of Wilma van Wezenbeek as the Executive Officer for FEMS Publications (2000-2006) and her assistant Gillian van Beest. Shortly afterwards, FEMS Executive Board invited me to set up its fifth journal, FEMS Yeast Research. An important task would be to find eligible candidates for the future editorial board. An excellent opportunity presented itself during the Tenth International Symposium on Yeasts, organized in 2000 at Papendal (Arnhem) (van Dijken & Scheffers, 2000). The invited candidates showed great approval and soon I could start with an outstanding editorial board. The new journal developed favourably, and after five years I
could turn over chief editorship to Teun Boekhout for another five years. Since then the task is in the able hands of Jens Nielsen. In 2003 I was honoured with the FEMS Special Award (Fig. 9). I enjoyed being involved in so many aspects of microbiology, in particular of yeasts. The concerted action with so many capable colleagues and friends was a great privilege and joy. I was happy to be a co-author in a collective paper (van Maris et al., 2006) by the group of Jack Pronk (Fig. 10). And I was delighted when our xylose-fermenting yeast, Pichia stipitis, was relegated to the new genus Scheffersomyces (Kurtzman & Suzuki, 2010). I congratulate FEMS at the occasion of its 40th anniversary and wish it a bright and golden future ! And may FEMS Yeast Research continue to be a valuable and esteemed offspring on its stock!
ACKNOWLEDGEMENT To Cocky, my wife, and our children Peter and Irene, for their love and support throughout the years.
REFERENCES Alonzo V & Scheffers WA (1975) Studies on hydrolysis and synthesis of fats in Staphylococcus aureus and Candida lipolytica. Boll Ist Sieroter Milanese 54: 90-97. Bakker G, Scheffers WA & Wikén TO (1968) A new method for the determination of clotting times in milk. Neth Milk and Dairy J 22: 16-21. Boekhout T, Renting M, Scheffers WA & Bosboom R (1993) The use of karyotyping in the systematics of yeasts. Antonie van Leeuwenhoek 63: 157-163. Boekhout T, van Belkum A, Leenders ACAP, Verbrugh HA, Mukamurangwa P, Swinne D & Scheffers WA (1997) Molecular typing of Cryptococcus neoformans: taxonomic and epidemiological aspects. Internatl. J Syst Bacteriol 47: 432-442. Bruinenberg PM, de Bot PHM, van Dijken JP & Scheffers WA (1983) The role of redox balances in the anaerobic fermentation of xylose by yeasts. Eur J Appl Microbiol Biotechnol 18: 287-292. Bruinenberg PM, de Bot PHM, van Dijken JP & Scheffers WA (1984) NADH-linked aldose reductase: the key to anaerobic alcoholic fermentation of xylose by yeasts. Appl Microbiol Biotechnol 19: 256-260. Bruinenberg PM, Jonker R, van Dijken JP & Scheffers WA (1985) Utilization of formate as an additional energy source by glucose-limited chemostat cultures of Candida utilis CBSB21 and Saccharomyces cerevisiae CBS8066. Evidence for the absence of transhydrogenase activity in yeasts. Arch Microbiol 142: 302-306. Bruinenberg PM, van Dijken JP, Kuenen JG & Scheffers WA(1985) Critical parameters in the isolation of mitochondria from Candida utilis grown in continuous culture. J Gen Microbiol 131: 1035-1042. Bruinenberg PM, van Dijken JP, Kuenen JG & Scheffers WA (1985) Oxidation of NADH and NADPH by mitochondria from the yeast Candida utilis. J Gen Microbiol 131: 1043-1051. Bruinenberg PM, van Dijken JP & Scheffers WA (1983) A theoretical analysis of NADPH production and consumption in yeasts. J Gen Microbiol 129: 953-964. Bruinenberg PM, van Dijken JP & Scheffers WA (1983) An enzymic analysis of NADPH production and consumption in Candida utilis. J Gen Microbiol 129: 965-971. Bruinenberg PM, van Dijken JP & Scheffers WA (1986) A radiorespirometric study on the contribution of the hexose monophosphate pathway to glucose metabolism in Candida utilisCB3621 grown in glucose-limited chemostat cultures. J Gen Microbiol 132: 221-229. Bruinenberg PM, Waslander GW, van Dijken JP & Scheffers WA (1986) A comparative radiorespirometric study of glucose metabolism in yeasts. Yeast 2: 117-121. Bulder CJEA. On Respiratory Deficiency in Yeasts. Ph.D. Thesis, Delft. Groningen: VRB, 1963.
Carrascosa JM, Viguera MD, Nuñez de Castro I & Scheffers WA (1981) Metabolism of acetaldehyde and Custers effect in the yeast Brettanomyces abstinens. Antonie van Leeuwenhoek 47: 209-215. Custers MTJ. Onderzoekingen over het Gistgeslacht Brettanomyces. Ph.D. Thesis, Delft. 1940. de Boer WE, Golten C & Scheffers WA (1975a) Effects of some physical factors on flagellation and swarming of Vibrio alginolyticus. Neth J Sea Res 9: 197-213. de Boer WE, Golten C & Scheffers WA (1975b) Effects of some chemical factors on flagellation and swarming of Vibrio alginolyticus. Antonie van Leeuwenhoek 41: 385-403. de Jonge P, de Jongh FCM, Meijers R, Steensma HY & Scheffers WA (1986) Orthogonal-fieldalternation gel electrophoresis patterns of DNA from yeasts. Yeast 2: 193-204. Dijkstra FIJ, Scheffers WA & Wikén TO (1972) Submerged growth of the cultivated mushroom, Agaricus bisporus. Antonie van Leeuwenhoek 38: 329-340. Golten C & Scheffers WA (1975) Marine vibrios isolated from water along the Dutch coast. Neth J Sea Res 9: 351-364. González FC, Scheffers WA & Mossel DAA (1971) Comparison of selective media for the enumeration of Lactobacillus species. Antonie van Leeuwenhoek 37: 262 264. Hanstveit AO, Gerritse GA & Scheffers WA. A study of the biodeterioration of vulcanized rubber sealings, exposed to inoculated tap water. In: Morton LHG (ed.).The Biodeterioration of Constructing Materials. Delft: The Biodeterioration Society, 1987, 87-96. Havelaar AH, Hoogendorp CJ, Wesdorp AJ & Scheffers WA (1980) False-negative oxidase reaction as a result of medium acidification. Antonie van Leeuwenhoek 46: 301-312. Hensing MCM, Rouwenhorst RJ, Scheffers WA & van Dijken JP (1993) Production and localization of inulinase in Kluyveromyces yeast. In: Fuchs A (ed.). Inulin and Inulin-Containing Crops. Amsterdam: Elsevier, 1993, 241-250. Hoogerheide JC. Bijdrage tot de Kennis van de Reactie van Pasteur. PhD Thesis, Delft, 1935. Houwink AL, le Poole JB & le Rütte WA (eds). Proceedings of the Conference on Electron Microscopy. Delft: Hooigland, 1950. Kamp AF, la Riviere JWM & Verhoeven W (eds). Albert Jan Kluyver. His Life and Work. Amsterdam: North-Holland Publ. Comp., 1959. Kluyver AJ. Biochemische Suikerbepalingen. PhD Thesis, Delft, 1914. Kluyver AJ & Donker HJL (1926) Die Einheit in der Biochemie. Chem Zelle Gewebe 13:134. Kreger-van Rij NJW. A Taxonomic Study of the Yeast Genera Endomycopsis, Pichia and Debaryomyces. Ph D Thesis, Delft, 1964. Kreger-van Rij NJW (ed.). The Yeasts. A Taxonomic Study. 3rd Ed. Amsterdam: Elsevier, 1984. Kurtzman CP & Suzuki M (2010) Phylogenetic analysis of ascomycete yeasts that form coenzyme Q-9 and the proposal of the new genera Babjeviella, Meyerozyma, Millerozyma, Priceomyces,and Scheffersomyces. Mycoscience 51: 2-14. Lodder J. Beiträge zu einer Monographie der Hefearten. II. Teil. Die anaskosporogenen Hefen. Erste Hälfte. PhD Thesis, Utrecht, 1934. Lodder J & Kreger-van Rij NJW. The Yeasts. A Taxonomic Study. Amsterdam: North Holland Publ. Comp., 1952. Nicolay K, Scheffers WA, Bruinenberg PM & Kaptein R (1982) Phosphorus -31 nuclear magnetic resonance studies of intracellular pH, phosphate compartmentation and phosphate transport in yeasts. Arch Microbiol 133: 83-89. Nicolay K, Scheffers WA, Bruinenberg PM & Kaptein R (1983) In vivo phosphorus-31 NMR studies on the role of the vacuole in phosphate metabolism in yeasts. Arch Microbiol 134: 270-275. Noshiro A, Purwin C, Laux M, Nicolay K, Scheffers WA & Holzer H (1987) Mechanism of stimulation of endogenous fermentation in yeast by carbonyl cyanide m-chlorophenyl hydrazone. J Biol Chem 262: 14154-14157. Pernice A, Alonzo V & Scheffers WA (1970) Activity of Staphylococcus aureus on some lipidic compounds of egg yolk. Atti Soc Peloritana Sc Fis Mat e Natur 16: 321 327. Postma E, Kuiper A, Tomasouw WF, Scheffers WA & van Dijken JP (1989) Competition for
glucose between the yeasts Saccharomyces cerevisiae and Candida utilis. Appl Environ Microbiol 55: 3214-3220. Postma E, Scheffers WA & van Dijken JP (1988) Adaptation of the kinetics of glucose transport to environmental conditions in the yeast Candida utilis CBS621: a continuous-culture study. J gen Microbiol 134:1109-1116. Postma E, Scheffers WA & van Dijken JP (1989) Kinetics of growth and glucose transport in glucose-limited chemostat cultures of Saccharomyces cerevisiae CBS8066. Yeast 5:159-165. Postma E, Verduyn C, Kuiper A, Scheffers WA & van Dijken JP (1990) Substrate-accelerated death of Saccharomyces cerevisiae CBS8066 under maltose stress. Yeast 6:149-158. Postma E, Verduyn C, Scheffers WA & van Dijken JP (1989) Enzymic analysis of the Crabtree effect in glucose-limited chemostat cultures of Saccharomyces cerevisiae. Appl Environ Microbiol 55: 468-477. Pronk JT, Bakker AW, van Dam HE, Straathof AJJ, Scheffers WA & van Dijken JP (1988) Preparation of D-xylulose from D-xylose. Enzyme Microb Technol 10:537-542. Pronk JT, van der Linden-Beuman A, Verduyn C, Scheffers WA & van Dijken JP (1994) Propionate metabolism in Saccharomyces cerevisiae: implications for the metabolon hypothesis. Microbiology 140: 717-722. Pronk JT, Wenzel TJ, Luttik MAH, Klaassen CCM, Scheffers WA, Steensma HY & van Dijken JP (1994) Energetic aspects of glucose metabolism in a pyruvate-dehydrogenase-deficient mutant of Saccharomyces cerevisiae. Microbiology 140: 601-610. Purwin C, Nicolay K, Scheffers WA & Holzer H (1986) Mechanism of control of adenylate cyclase activity in yeast by fermentable sugars and carbonyl cyanide m-chlorophenyl hydrazone. J Biol Chem 261: 8744-8749. Rouwenhorst RJ, Frank J, Scheffers WA & van Dijken JP (1991) Determination of protein concentration by total carbon analysis (1991) J Biophys Methods 22:119-128. Rouwenhorst RJ, Hensing M, Verbakel J, Scheffers WA & van Dijken JP (1990) Structure and properties of the extracellular inulinase of Kluyveromyces marxianus CBS6556. Appl Environ Microbiol 56:3337-3345. Rouwenhorst RJ, Ritmeester WS, Scheffers WA & van Dijken JP (1990) Localization of inulinase and invertase in Kluyveromyces species. Appl Environ Microbiol 56: 3329-3336. Rouwenhorst RJ, van der Baan AA, Scheffers WA & van Dijken JP (1991) Production and localization of B-fructosidase in asynchronous and synchronous cultures of yeasts. Appl Environ Microbiol 57:557-562. Rouwenhorst RJ, Visser LE, van der Baan AA, Scheffers WA & van Dijken JP (1988) Production, distribution, and kinetic properties of inulinase in continuous cultures of Kluyveromyces marxianus CBS 6556.Appl Environ Microbiol 54: 1131-1137. Scheffers WA (1961) On the inhibition of alcoholic fermentation in Brettanomyces yeasts under anaerobic conditions. Experientia 17:40-42. Scheffers WA (1966) Stimulation of fermentation in yeasts by acetoin and oxygen. Nature 210: 533534. Scheffers WA (1967) Effects of oxygen and acetoin on fermentation and growth in Brettanomyces and some other yeast genera. Atti del XIV Congr Soc Ital Microbiol: 91-107. Scheffers WA (1994) The Netherlands Society for Microbiology/ Nederlandse Vereniging voor Microbiologie (NvvM). SGM Quarterly: 77-78. Scheffers WA . Honderd jaar leerstoel microbiologie Delft, 1895-1995. In: Jaarboek Nederlandse Vereniging voor Microbiologie 1996, 11-15. Scheffers WA (1999) Eijkman Centennial on Infections in the 21st Century. “Successes from the past, challenges for the future”. FEMS Immunol Medic Microbiol 26: 183-321. Scheffers WA & Misset O (1974) The Custers effect in Brettanomyces intermedius. Proc. 4 th Int Symp on Yeasts, Vienna, Part I: 39-40. Scheffers WA, te Nijenhuis B & Kragten J (eds) (1984) Analytical Methods and Problems in Biotechnology. Proceed Internatl Symp, Noordwijkerhout. Analytica Chim Acta 163: 1-328.
Scheffers WA & van Dijken JP (eds). Metabolic Compartmentation in Yeasts. 16th Internatl Specialized Symp on Yeasts (ISSY 16, Arnhem,1993). The Hague: Pasmans, 1993. Scheffers WA & van Dijken JP (eds). Beijerinck Centennial. Microbial Physiology and Gene Regulation: Emerging Principles and Applications. Delft: Delft Univ. Press, 1995. Scheffers WA & Wikén TO (1969) The Custers effect (negative Pasteur effect) as a taxonomic criterion for the genus Brettanomyces. Antonie van Leeuwenhoek 35: A31-A32. Smith MT, Batenburg-van der Vegte WH & Scheffers WA (1981) Eeniella, a new yeast genus of the Torulopsidales. Internatl J Syst Bacteriol 31:196-203. Stelling-Dekker NM. Beiträge zu einer Monographie der Hefearten. I. Teil. Die Sporogenen Hefen. PhD Thesis, Utrecht, 1931. Struyk AP. Onderzoekingen over de Alcoholische Gisting. PhD Thesis, Delft, 1928. te Nijenhuis B, van Dijk C, Kragten J & Scheffers WA (eds) (1988) Analytical Methods and Problems in Biotechnology. Proceed 2 nd Internatl Symp, Anabiotec'88, Noordwijkerhout. Analytica Chim Acta 213:1-183. Toivola A, Yarrow D, van den Bosch E, van Dijken JP & Scheffers WA (1984) Alcoholic fermentation of D-xylose by yeasts. Appl Environ Microbiol 47:1221-1223. Tromp AGGM, Bonnet JABAF & Scheffers WA (1968) A simple culture vessel for turbidimetric studies of anaerobic growth of microbes. Experientia 24:1076-1077. Tromp AGGM & Scheffers WA (1969) Effect of acetoin on anaerobic growth in Brettanomyces. Antonie van Leeuwenhoek 35: H37-H38. van der Walt JP. On the Yeast Candida pulcherrima and its Pigment. PhD Thesis, Delft. van Dijken JP & Scheffers WA. Studies on alcoholic fermentation in yeasts. In: Houwink EH & van der Meer (eds). Innovations in Biotechnology. Amsterdam: Elsevier Science Publ, 1984. van Dijken JP & Scheffers WA (1985) Procedé de production d' éthanol à partir d' une substance contenant du xylose. Patent: FR 2545838 (A1). van Dijken JP & Scheffers WA (1986) Redox balances in the metabolism of sugars by yeasts. FEMS Microbiol Reviews 32: 199-224. van Dijken JP & Scheffers WA (eds) The Rising Power of Yeasts in Science and Industry. ISY 2000. Tenth Internatl. Symp. On Yeasts, Arnhem, The Netherlands. Delft University Press, 2000. van Dijken JP, van den Bosch E, Hermans JJ, Rodrigues de Miranda L & Scheffers WA (1986) Alcoholic fermentation by “non-fermentative”yeasts. Yeast 2: 123-127. van Doorne H, Scheffers WA, Hadiutomo M & van den Bosch E (1984) Microbial contamination of a vitamin A formulation, prepared in local pharmacies, and its preservation against yeasts and moulds. Antonie van Leeuwenhoek 50: 405-416. van Maris AJA, Abbott DA, Bellissimi E, van den Brink J, Kuyper M, Luttik MAH, Wisselink HW, Scheffers WA, van Dijken JP & Pronk JT (2006) Alcoholic fermentation of carbon sources in biomass hydrolysates by Saccharomyces cerevisiae: current status. Antonie van Leeuwenhoek 90: 391-418. van Urk H, Bruinenberg PM, Veenhuis M, Scheffers WA & van Dijken JP (1989) Respiratory capacities of mitochondria of Saccharomyces cerevisae CBS 8066 and Candida utilis CBS 621 grown under glucose limitation. Antonie van Leeuwenhoek 56: 211-220. van Urk H, Mak PR, Scheffers WA & van Dijken JP (1988) Metabolic responses of iSaccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621upon transition from glucose limitation to glucose excess. Yeast 4: 283-291. van Urk H, Postma E, Scheffers WA & van Dijken JP (1989) Glucose transport in Crabtree-positive and Crabtree-negative yeasts. J Gen Microbiol 135: 2399-2406. van Urk, H, Schipper D, Breedveld GJ, Mak PR, Scheffers WA & van Dijken JP (1989) Localization and kinetics of pyruvate-metabolizing enzymes in relation to aerobic alcoholic fermentation in Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621. Biochim Biophys Acta 992: 78-86. van Urk H, Voll WSL, Scheffers WA & van Dijken JP (1990) Transient-state analysis of metabolic fluxes in Crabtree-positive and Crabtree-negative yeasts. Appl Environ Microbiol.56:281-287.
Verduyn C, Breedveld GJ, Scheffers WA & van Dijken JP (1988) Purification and properties of dihydroxyacetone reductase and 2,3-butanediol dehydrogenase from Candida utilis CBS 621. Yeast 4: 127-133. Verduyn C, Breedveld GJ, Scheffers WA & van Dijken JP (1988) Metabolism of 2-3-butanediol in yeasts. Yeast 4: 135-142. Verduyn C, Breedveld GJ, Scheffers WA & van Dijken JP (1988) Substrate specificity of alcohol dehydrogenase from the yeasts Hansenula polymorpha CBS 4732 and Candida utilis CBS 621. Yeast 4:143-148. Verduyn C, Breedveld GJ, Schreuder H, Scheffers WA & van Dijken JP (1988) Properties of enzymes which reduce dihydroxyacetone and related compounds in Hansenula polymorpha CBS 4732. Yeast 4: 117-126. Verduyn C, Frank J, van Dijken JP & Scheffers WA (1985) Multiple forms of xylose reductase in Pachysolen tannophilus. FEMS Microbiol Lett 30: 313-317. Verduyn C, Giuseppin MLF, Scheffers WA & van Dijken JP (1988) Hydrogen peroxide metabolism in yeasts. Appl. Environ. Microbiol. 54: 2086-2090. Verduyn C, Postma E., Scheffers WA & van Dijken JP (1990) Physiology of Saccharomyces cerevisiae in anaerobic glucose-limited chemostat cultures. J Gen Microbiol 136: 395-403. Verduyn C, Postma E, Scheffers WA & van Dijken JP (1990) Energetics of Saccharomyces cerevisiae in anaerobic glucose-limited chemostat cultures. J Gen Microbiol 136: 405-412. Verduyn C, Postma E, Scheffers WA & van Dijken JP (1992) Effect of benzoic acid on metabolic fluxes in yeasts: a continuous-culture study on the regulation of respiration and alcoholic fermentation. Yeast 8: 501-517. Verduyn C, Stouthamer AH, Scheffers WA & van Dijken JP (1991) A theoretical evaluation of growth yields of yeasts. Antonie van Leeuwenhoek 59: 49-63. Verduyn C, van Dijken JP & Scheffers WA (1983) A simple, sensitive, and accurate alcohol electrode. Biotechnol Bioeng 25: 1049-1055. Verduyn C, van Dijken JP & Scheffers WA (1984) Colorimetric alcohol assays with alcohol oxidase. J Microbiol Methods 2: 15-25. Verduyn C, van Kleef R, Frank J, Schreuder H, van Dijken JP & Scheffers WA (1985) Properties of the NAD(P)H-dependent xylose reductase from the xylose-fermenting yeast Pichia stipitis. Biochem J 226: 669-677. Verduyn C, van Wijngaarden CJ, Scheffers WA & van Dijken JP (1991) Hydrogen peroxide as an electron acceptor for mitochondrial respiration in the yeast Hansenula polymorpha. Yeast 7: 137146. Verduyn C, Zomerdijk TPL, van Dijken JP & Scheffers WA (1984) Continuous measurement of ethanol production by aerobic yeast suspensions with an enzyme electrode. ApplMicrobiol Biotechnol 19: 181-185. Visser W, Scheffers WA, Batenburg-van der Vegte WH & van Dijken JP (1990) Oxygen requirements of yeasts. Appl Environ Microbiol 56: 3785-3792. Visser W, van der Baan AA, Batenburg-van der Vegte WH, Scheffers WA, Krämer R & van Dijken JP (1994) Involvement of mitochondria in the assimilatory metabolism of anaerobic Saccharomyces cerevisiae cultures. Microbiology 140: 3039-3046. Weusthuis RA, Adams H, Scheffers WA & van Dijken JP (1993) Energetics and kinetics of maltose transport in Saccharomyces cerevisiae: a continuous-culture study. Appl Environ Microbiol 59: 3102-3109. Weusthuis RA, LuttikMAH, Scheffers WA, van Dijken JP & Pronk JP (1994) Is the Kluyver effect in yeasts caused by product inhibition? Microbiology 140: 1723-1729. Weusthuis RA, Visser W, Pronk JP, Scheffers WA & van Dijken JP (1994) Effects of oxygen limitation on sugar metabolism in yeasts: a continuous-culture study of the Kluyver effect. Microbiology 140:703-715. Wijsman MR, van Dijken JP, van Kleeff BHA& Scheffers WA (1984) Inhibition of fermentation and growth in batch cultures of the yeast Brettanomyces intermedius upon a shift from aerobic to
anaerobic conditions (Custers effect). Antonie van Leeuwenhoek 50: 183-192. Wikén T, Scheffers WA & Verhaar AJM (1961) On the existence of a negative Pasteur effect in yeasts classified in the genus Brettanomyces Kufferath et van Laer. Antonie van Leeuwenhoek 27: 401-433. Wiken T, Verhaar AJM & Scheffers WA (1962) The influence of potassium and sodium ions on the negetive Pasteur effect in Brettanomyces claussenii Custers. Archiv f. Mikrobiol. 42: 226-236.
FIGURE LEGENDS
Fig.1 The Professor’s residence (left part) connected to the laboratory (right) of Beijerinck and Kluyver at Nieuwelaan 3
Fig.2 Kluyver in the library. Photo by J.A. Schuur
Fig.3 Kluyver in his workroom with van Niel, during the preparation of their book (1955). Photo by J.K. Baars
Fig. 4 The author at a manometric experiment, measuring aerobic and anaerobic fermentation and respiration rates of yeasts (1958)
Fig. 5 The laboratory of Julianalaan
Fig. 6 Wikén with the author. Photo by J.A. Schuur
Fig. 7 Nel Kreger-van Rij, Yeast Division of Centraalbureauu voor Schimmelcultures (CBS)
Fig. 8 Gijs Kuenen with Peter Arntz, at the latter’s silver jubilee (1980). Photo by J.A. Schuur
Fig. 9 FEMS Special Award, presented to the author during the first FEMS Symposium, Ljubljana (2003). Photo by Diman van Rossum
Fig. 10 The author, Jack Pronk and Hans van Dijken (from left to right) on the lake of Bled, Slovenia, during the International Specialised Symposium on Yeasts (1997). Photo by Leonie Raamsdonk
