Archives of
Microbiology
Arch. Microbiol. 107, 205-206 (1976)
9 by Springer-Verlag 1976
Torulopsis spandovensis sp. n., WOLFGANG HENNINGER
a N e w Yeast from Beer
a~d S I E G F R I E D W I N D I S C H
Deutsche Sammlung von Mikroorganismen, Teilsammlung Berlin II, im Institut ffir Gfirungsgewerbe und Biotechnologie, Seestr. 13, D-1000 Berlin 65
Abstract. A new species of the genus Torulopsis has been isolated from several different samples of G e r m a n Pilsener Beer. A description of the new species is given.
Key words." Torulopsis -
Yeast -
Dalmau Plate Culture on Potato-Glucose Agar There was no formation of pseudomycelium neither under aerobic nor anaerobic conditions.
New species -
Beer.
During an investigation on the qualitative and quantitative occurrence of yeasts in a large number of G e r m a n beers two strains of an undescribed Torulopsis species were detected. The yeast is a weak fermenter and resembles T. apis Lavie ex van Uden et Vidal-Leiria (1970) closely. METHODS The methods used were those as described by van der Walt 0970). The media for assimilation tests were solMified. Instead of cornmeal agar potato-glucose agar was used for Dalmau plate cultures.
Formation of Ascospores Ascospores were not observed on malt-extract agar nor on the various sporulation media proposed by Fowell (1952) and McClary et al. (1959).
Fermentation Glucose Galactose Sucrose Maltose Cellobiose Trehalose Lactose
+ (slow) + (slow, weak) + ( s l o w , weak) -
Melibiose Raffinose Melezitose Inulin Soluble starch c~-MethylD-glucoside
--
Assimilation of carbon Compounds RESULTS
Growth on Malt-Extract Agar After 3 days of incubation at 28 ~C the cells were round or shortly oval ( 2 . 8 - 6 . 0 ) x ( 2 . 8 - 6 . 0 ) g i n . Propagation occurred by multilateral budding. The cells occurred singly, occasionally in pairs and in small clusters. The streak culture was butyrous, creamcolored, flat, shiny and smooth. The margin was entire. The appearance did not change after several weaks of cultivation. The culture produced a slight fruity odour.
Growth in Malt-Extract Solution The cells resembled those on malt-extract agar. The solution was clear, a sediment was formed, after 3 days a thin ring appeared.
Glucose Galactose L-Sorbose Sucrose Maltose Cellobiose Trehalose Lactose Melibiose Raffinose Melezitose Inulin Soluble starch D-Xylose L-Arabinose D-Arabinose D-Ribose
+ + + + -
+ +/+/+/-
L-Rhamnose Ethanol Glycerol Erythritol Ribitol Galactitol D-Mannitol D-Glucitol c~-MethylD-glucoside Salicin DE-Lactic acid Succinic acid Citric acid Inositol EthylamineHC1
+/+ + + + + -+/+ +/-
+
206 The cells grew on 6 0 ~ glucose containing yeast extract agar, tolerated 8 - 9 ~ NaCl, were resistant to 25 gg cycloheximide/ml, grew at 37~ did not grow on vitamin-free medium, did not use KNO3, starch-like compounds were not formed, pyridoxine had a stimulating effect. Two strains were collected from Pilsener type of German beer. The two strains were deposited at the Deutsche Sammlung von Mikroorganismen. DSM 70866 is regarded to be the type strain. The name T. spandovensis derives from the Latin word for the old town of Spandau (Spandove), now a district of Berlin, where one of the two strains has been isolated.
DISCUSSION Extensive research was done to characterize the biological composition of German beer at various stages during its production (Windisch, 1962). Fermenting and non-fermenting yeasts were found to occur in different amounts as well as in varying ratios. Two strains were found among the yeasts which apparently belonged to the same, yet hitherto undescribed species. We have investigated the characteristics of this yeast and named it Torutopsis spandovensis. The assimilation profile of T. spandovensis resembles that of T. apis Lavie ex van Uden et VidalLeiria, although the former can also assimilate L-SOtbose, ribitol, and galactitol. The two species also differ with regard to their fermentation properties: T. apis cannot ferment at all, whereas T. spandovensis is able to ferment glucose, as well as galactose and sucrose slightly. In contrast to T. spandovensis, T. apis can grow without the addition of external vitamins. Although both yeasts seem to be closely related, it is improbable that one species has evolved directly out of the other. It is conceivable, that both species have evolved from a third, as yet unknown species by loss of one or the other gene function. The postulated species must have possessed (or possesses) the ability to ferment the various sugars and to assimilate the other carbon sources as does T. spandovensis, as well as the ability to synthesize vitamins, as does
T. apis.
Arch. Microbiol.,Vol. 107, No. 2 (1976) LATIN DIAGNOSIS
Torulopsis spandovenis sp. n. In extracto maltato cellulae rotundae, sphaeroidales, ovoideae ( 2 . 8 - 6 . 0 ) x ( 2 . 8 - 6 . 0 ) gm, singulae raro binae aut in catenis. Sedimentum et annulus subtilis formantur. In agaro maltato cellulae ut in medio liquido. Colonia in medio solido glabra.~ nitida, margine glabra. Pseudomycelium nullum. Ascosporae desunt. Glucosum, galactosum (lente) et sucrosum (lente) fermentantur ac non raffinosum. Glucosum, galactosum, L-sorbosum, sucrosum, raffinosum, glycerolum, ribitolum, galactitolum, Dmannitolum, D-glucitolum, acidum succinicum et aethylaminum hydrochloricum assimilantur at non maltosum, cellobiosum, trehalosum, lactosum, melibiosum, melezitosum, amylum solubile, D-arabinosum, D-ribosum, L-rhamnosum, i-erythritolum, a-methylD-m-glucosidum, salicinum, i-inositolum. Assimilationes inulini, D-xylosi, L-arabinosi, aethanoli, DL-acidi lactici et acidi citrici variant. Kalium nitricum assimilatur. Ad crescentiam vitaminae externae necesse sunt. Crescit in 37 ~C. Materia amyloidea non formatur.
Acknowledgements. This work was supported by the Bundesministerium fiir Forschung und Technologie(BCT 85).
REFERENCES Fowell, R. R. : Sodium acetate agar as a sporulation medium for yeast. Nature (Lond.) 170, 578 (1952) McClary, D. O., Nulty, W. L., Miller, G. R. : Effect of potassium versus sodium in the sporulation of Saccharomyces. J. Bact. 78, 362- 368 (1959) van Uden, N., Vidal-Leiria,M. : TorulopsisBerlese. In: The yeasts (J. Lodder, ed.), pp. 1235-1308, 2nd ed. Amsterdam-London: North Holland 1970 van der Walt, J. P. : Criteria and methods used in classification. In: The yeasts (J. Lodder, ed.), pp. 34-113, 2nd ed. Amsterdam-London: North-Holland 1970 Windisch, S. : Ober die Ergebnisse weiterer vegetationskundlicher Analysen an Bieren. Mschr. Brauerei 15, 203-210 (1962)
Received October 13, 1975
Microbiology
Arch. Microbiol. 107, 205-206 (1976)
9 by Springer-Verlag 1976
Torulopsis spandovensis sp. n., WOLFGANG HENNINGER
a N e w Yeast from Beer
a~d S I E G F R I E D W I N D I S C H
Deutsche Sammlung von Mikroorganismen, Teilsammlung Berlin II, im Institut ffir Gfirungsgewerbe und Biotechnologie, Seestr. 13, D-1000 Berlin 65
Abstract. A new species of the genus Torulopsis has been isolated from several different samples of G e r m a n Pilsener Beer. A description of the new species is given.
Key words." Torulopsis -
Yeast -
Dalmau Plate Culture on Potato-Glucose Agar There was no formation of pseudomycelium neither under aerobic nor anaerobic conditions.
New species -
Beer.
During an investigation on the qualitative and quantitative occurrence of yeasts in a large number of G e r m a n beers two strains of an undescribed Torulopsis species were detected. The yeast is a weak fermenter and resembles T. apis Lavie ex van Uden et Vidal-Leiria (1970) closely. METHODS The methods used were those as described by van der Walt 0970). The media for assimilation tests were solMified. Instead of cornmeal agar potato-glucose agar was used for Dalmau plate cultures.
Formation of Ascospores Ascospores were not observed on malt-extract agar nor on the various sporulation media proposed by Fowell (1952) and McClary et al. (1959).
Fermentation Glucose Galactose Sucrose Maltose Cellobiose Trehalose Lactose
+ (slow) + (slow, weak) + ( s l o w , weak) -
Melibiose Raffinose Melezitose Inulin Soluble starch c~-MethylD-glucoside
--
Assimilation of carbon Compounds RESULTS
Growth on Malt-Extract Agar After 3 days of incubation at 28 ~C the cells were round or shortly oval ( 2 . 8 - 6 . 0 ) x ( 2 . 8 - 6 . 0 ) g i n . Propagation occurred by multilateral budding. The cells occurred singly, occasionally in pairs and in small clusters. The streak culture was butyrous, creamcolored, flat, shiny and smooth. The margin was entire. The appearance did not change after several weaks of cultivation. The culture produced a slight fruity odour.
Growth in Malt-Extract Solution The cells resembled those on malt-extract agar. The solution was clear, a sediment was formed, after 3 days a thin ring appeared.
Glucose Galactose L-Sorbose Sucrose Maltose Cellobiose Trehalose Lactose Melibiose Raffinose Melezitose Inulin Soluble starch D-Xylose L-Arabinose D-Arabinose D-Ribose
+ + + + -
+ +/+/+/-
L-Rhamnose Ethanol Glycerol Erythritol Ribitol Galactitol D-Mannitol D-Glucitol c~-MethylD-glucoside Salicin DE-Lactic acid Succinic acid Citric acid Inositol EthylamineHC1
+/+ + + + + -+/+ +/-
+
206 The cells grew on 6 0 ~ glucose containing yeast extract agar, tolerated 8 - 9 ~ NaCl, were resistant to 25 gg cycloheximide/ml, grew at 37~ did not grow on vitamin-free medium, did not use KNO3, starch-like compounds were not formed, pyridoxine had a stimulating effect. Two strains were collected from Pilsener type of German beer. The two strains were deposited at the Deutsche Sammlung von Mikroorganismen. DSM 70866 is regarded to be the type strain. The name T. spandovensis derives from the Latin word for the old town of Spandau (Spandove), now a district of Berlin, where one of the two strains has been isolated.
DISCUSSION Extensive research was done to characterize the biological composition of German beer at various stages during its production (Windisch, 1962). Fermenting and non-fermenting yeasts were found to occur in different amounts as well as in varying ratios. Two strains were found among the yeasts which apparently belonged to the same, yet hitherto undescribed species. We have investigated the characteristics of this yeast and named it Torutopsis spandovensis. The assimilation profile of T. spandovensis resembles that of T. apis Lavie ex van Uden et VidalLeiria, although the former can also assimilate L-SOtbose, ribitol, and galactitol. The two species also differ with regard to their fermentation properties: T. apis cannot ferment at all, whereas T. spandovensis is able to ferment glucose, as well as galactose and sucrose slightly. In contrast to T. spandovensis, T. apis can grow without the addition of external vitamins. Although both yeasts seem to be closely related, it is improbable that one species has evolved directly out of the other. It is conceivable, that both species have evolved from a third, as yet unknown species by loss of one or the other gene function. The postulated species must have possessed (or possesses) the ability to ferment the various sugars and to assimilate the other carbon sources as does T. spandovensis, as well as the ability to synthesize vitamins, as does
T. apis.
Arch. Microbiol.,Vol. 107, No. 2 (1976) LATIN DIAGNOSIS
Torulopsis spandovenis sp. n. In extracto maltato cellulae rotundae, sphaeroidales, ovoideae ( 2 . 8 - 6 . 0 ) x ( 2 . 8 - 6 . 0 ) gm, singulae raro binae aut in catenis. Sedimentum et annulus subtilis formantur. In agaro maltato cellulae ut in medio liquido. Colonia in medio solido glabra.~ nitida, margine glabra. Pseudomycelium nullum. Ascosporae desunt. Glucosum, galactosum (lente) et sucrosum (lente) fermentantur ac non raffinosum. Glucosum, galactosum, L-sorbosum, sucrosum, raffinosum, glycerolum, ribitolum, galactitolum, Dmannitolum, D-glucitolum, acidum succinicum et aethylaminum hydrochloricum assimilantur at non maltosum, cellobiosum, trehalosum, lactosum, melibiosum, melezitosum, amylum solubile, D-arabinosum, D-ribosum, L-rhamnosum, i-erythritolum, a-methylD-m-glucosidum, salicinum, i-inositolum. Assimilationes inulini, D-xylosi, L-arabinosi, aethanoli, DL-acidi lactici et acidi citrici variant. Kalium nitricum assimilatur. Ad crescentiam vitaminae externae necesse sunt. Crescit in 37 ~C. Materia amyloidea non formatur.
Acknowledgements. This work was supported by the Bundesministerium fiir Forschung und Technologie(BCT 85).
REFERENCES Fowell, R. R. : Sodium acetate agar as a sporulation medium for yeast. Nature (Lond.) 170, 578 (1952) McClary, D. O., Nulty, W. L., Miller, G. R. : Effect of potassium versus sodium in the sporulation of Saccharomyces. J. Bact. 78, 362- 368 (1959) van Uden, N., Vidal-Leiria,M. : TorulopsisBerlese. In: The yeasts (J. Lodder, ed.), pp. 1235-1308, 2nd ed. Amsterdam-London: North Holland 1970 van der Walt, J. P. : Criteria and methods used in classification. In: The yeasts (J. Lodder, ed.), pp. 34-113, 2nd ed. Amsterdam-London: North-Holland 1970 Windisch, S. : Ober die Ergebnisse weiterer vegetationskundlicher Analysen an Bieren. Mschr. Brauerei 15, 203-210 (1962)
Received October 13, 1975
