INTERNATIONAL
JOURNAL
OF SYSTEMATIC BACTERIOLOGY,
Jan. 1980,p. 196-205
0020-77 13/80/01-0196/10$02.OO/0
Vol. 30, No. 1
Synonymy of Saccharomyces cereuzsiae Hansen 1883 and
Saccharomyces uuarum Beijerinck 1898: Significance of Cell
Wall Antigens in Yeast Classification
Y. FUKAZAWA,' T. SHINODA,' A. NISHIKAWA,' A N D T. NAKASE'
Department of Microbiology, Me& College of Pharmacy, Tokyo 154, Japan,' and Research Institute for Life
Sciences, Ajinomoto Company, Yokohama 244, Japan'
Antigenic differences among strains of Saccharomyces cereuisiae Hansen 1883
and Saccharomyces uvarum Beijerinck 1898 were analyzed with respect to the
proton magnetic resonance spectra of alkali-extracted mannans and the gel
filtration profdes of the oligosaccharides of these mannans obtained by acetolysis.
The strains of both species could be divided into serotypes Ia, Ib, and 11. The
serological relationships of these strains were found to correlate with the proton
magnetic resonance spectra of the cell wall mannans. The gel filtratin profiles of
the acetolysis oligosaccharides of type Ia, Ib, and I1 strains of both species were
also characteristic for each type or subtype. Based on these findings, it is proposed
that S. cerevisiae and S. uvarum be combined into one species with two main
serotypes. S. uvarum is thus a later subjective synonym of S. cerevisiae.
Tsuchiya et al. (16,20,21,23) investigated the
serological relationships among strains of Saccharomyces cerevisiae, Saccharomyces uvarum,
Saccharomyces carlsbergensis, and other Saccharomyces species and showed that the structures of the heat-stable antigens of S. cerevisiae
are identical to those of S. uvarum but not to
those of S. carlsbergensis. Campbell and Allan
(2) reported that S. cerevisiae could be distinguished from S. cereuisiae var. ellipsoideus by
the serological method. Later, Campbell and
Brudzynski (3) reported that there are two antigenically distinct subgroups of S. carlsbergensis. Furthermore, Campbell (1) demonstrated
that the second antigenic group of S. carlsbergensis is antigenically similar or identical to
strains of S. uvarum. Richards (14) used an
immunofluorescent technique and found heterogeneous but closely related serotypes among
strains of S. cereuisiae, S. carlsbergensis, and S.
uuarum.
Gorin and Spencer (8) suggested the heterogeneity of strains of S. cerevisiae and S. carlsbergensis on the basis of the proton magnetic
resonance (PMR) spectra of alkali-extracted
mannans. Spencer et al. (15) also analyzed the
genetic control of the two types of mannans
produced by S. cerevisiae. Furthermore, Cawley
and Ballou (4) reported two structurally and
antigenically different S. cerevisiae cell wall
mannans.
S. carlsbergensis was placed into the species
S. uuarum by van der Walt (25).
In this paper, we report on an examination of
strains of S. cerevisiae and S. uvarum with
respect to the PMR spectra of alkali-extracted
mannans and the gel filtration profiles of the
oligosaccharides of these acetolysis-obtained
mannans so as to elucidate the antigenic differences among these strains.
MATERIALS AND METHODS
Strains used. Four strains of S. cereuisiae and
eight strains of S. uuarum were used as antigens for
the serological tests, the PMR spectra, and the gel
filtration profiles of acetolysis polysaccharides. The
strains are listed in Table 1, together with the sources
from which they were obtained.
Biological characteristics. The taxonomic characteristics of the strains were determined by the methods described by Wickerham (26) and van der Walt
(25).
Extraction and purification of polysaccharides. Yeast cells grown on Sabouraud medium for 48
h were harvested, washed three times with distilled
water, and heated at 100°C for 2 h. After centrifugation, the cells were suspended in 5 volumes of 2%
aqueous potassium hydroxide solution for 2 h at 100°C.
Subsequent extraction and purification of polysaccharides were done according to the methods of Gorin
and Spencer (7).
Determination of PMR spectra of polysaccharides. PMR spectra were determined for 20% solutions
of polysaccharides in deuterium oxide at 70°C with a
JEOL 100-MHz nuclear magnetic resonance spectrometer (Japan Electron Optics, Tokyo, Japan); sodium 2,2-dimethyl-2-silapentane-5-sulfonate
was employed as the internal standard.
Serological analyses. Serological characteristics
of the strains were determined by slide agglutination
tests with factor sera or absorbed antisera. Antisera to
strains were prepared by the method of Tsuchiya et
196
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VOL. 30,1980
SYNONYMY OF S. CEREVISIAE AND S. UVARUM
charide was dissolved in 10.5 ml of acetolysis medium
(a mixture of acetic acid, acetic anhydride, and concentrated sulfuric acid [lO:lO:l, vol/vol]). The solution
was warmed in an oil bath at 40°C. To obtain complete
cleavage of a-(1+6) linkages, acetolysis was carried
out for 13 h. The reaction was stopped by adding 2
volumes of anhydrous pyridine to the solution. The
solvent was evaporated, the oily residue was extracted
with 50 ml of chloroform-water (l:l, vol/vol), and the
chloroform extract was evaporated to dryness. The
residue was dissolved in 2 ml of anhydrous methanol,
and sodium methoxide in methanol was added dropwise until the solution became alkaline. After 20 min.
the solution was centrifuged, and the precipitate was
washed with dried methanol and dissolved in 2 ml of
water. The solution was neutralized by adding Amberlite IR 120. Deacetylated acetolysis products were
separated by gel filtration.
Column chromatography o n Bio-Gel P-2.The
deacetylated acetolysis products from 100 mg of mannan were applied to a P-2 gel column (2.5 by 150 cm).
The column was eluted with water at the rate of 20
ml/h at room temperature. Total carbohydrate in the
effluents was determined by the phenol-sulfuric acid
method (13) and the Fiske-Subbarow method for
phosphorus determination, as modified by Furukawa
et al. (6).
al. (18). Yeast cells grown on Sabouraud medium at
27°C for 48 h were harvested with saline solution,
heated at 100°C for 2 h, washed three times with 0.5%
formalinized saline solution, and adjusted to McFarland scale no. 9. Rabbits were injected intravenously with 0.5, 1.0, 2.0, 4.0, 4.0, and 4.0 ml of cell
suspensions at 4-day intervals. Factor sera 1,4,5,6,8,
9, 11, 13, 13b, and 34, prepared by the method of
Fukazawa et al. (5) for the diagnosis of medically
important yeasts, were obtained from Iatron Laboratory, Tokyo, Japan. Factor sera 10 and 18 were prepared by the method of Tsuchiya et al. (16-18). Each
antiserum to be absorbed was diluted to obtain an
agglutinin titer of approximately 1:256. For example,
when the agglutinin titer was 1:2,560, the antiserum
was diluted 10 times. A 2-ml amount of diluted serum
and 1ml of packed wet cells were mixed and agitated
for 2 h at 37°C and left standing overnight at 4°C. The
suspension was then centrifuged, and the supernatant
was tested against absorbed antigen by the slide agglutination test.
Quantitative precipitation tests. Quantitative
precipitin reactions between antisera and mannan antigens were performed in physiological saline solution,
with 0.1 ml of serum in a final volume of 1.0 ml for
both S. cerevisiae and S. uvarum strains. The reaction
mixture was incubated at 4°C for 48 h, and the precipitate was washed twice with ice-cold physiological
saline solution by centrifugation. Protein was measured by the method of Lowry et d. (11).
Acetylation and acetolysis of mannan. Acetylation and acetolysis were carried out by the method
of Kocourek and Ballou (9). Polysaccharides were
acetylated in a 1:l anhydrous pyridine-acetic anhydride mixture (100 mg of polysaccharide in 10 ml of
the mixture) by heating in a steam bath for 8 h, and
the solvent was evaporated. The acetylated polysac-
TABLE1. Sources of strains
Strain"
Type
197
RESULTS
Biological characteristics. The assimilation and fermentation patterns of carbon compounds by the four strains of S. cerevisiae and
eight strains of S. uvarum are shown in Table 2.
The characteristics of these strains correlated
well with the standard descriptions of their reof
S. cerevisiae and S. uvarum
Source or commenta
Ia
S. cerevisiae
S. cerevisiae
S. cerevisiae
S, uvarum
S. uvarum
M 6001
AJ 4001
AJ 4002
AJ 5286
I F 0 0615
I F 0 0209
IAM 4512
IAM 4274
(S.carkbergensis), AJ stock strain
Type strain (= CBS 395)
Ib
S. uvarum
S. uvarum
S. uvarum
I F 0 0565
I F 0 0751
AJ 4033
(S.carlsbergensis), FEKU, ATCC 9080
(S. carlsbergensis), NRRL 379
(S. carkrbergensis), IAM 4206
I1
S. cerevisiae
S. uvarum
S. uvarum
S. uvarum
CBS 1171
CBS 1513
I F 0 0220
I F 0 0297
Type strain
(S. carlsbergensis type strain)
(S. monacensis), CLMR, NCTC
(S.monacensis), HUT, NCTC
a The source abbreviations and the letters in the strain designations indicate the following institutions: M,
Meiji College of Pharmacy, Tokyo, Japan; AJ, Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki,
Japan; IFO, Institute for Fermentation, Osaka, Japan; CBS, Centraalbureau voor Schimmelcultures, Baarn,
The Netherlands, IAM, Institute of Applied Microbiology, University of Tokyo, Tokyo, Japan; ATCC, American
Type Culture Collection, Rockville, Md.; NRRL, Agricultural Research Service Culture Collection, Northern
Regional Research Center, U S . Department of Agriculture, Peoria, Ill.; CLMR, Central Laboratory, South
Manchuria Railway Co., Ltd. (defunct); NCTC, National Collection of Type Cultures, Central Public Health
Laboratory, London, England; HUT, Faculty of Engineering, Hiroshima University, Hiroshima, Japan; FEKU,
Faculty of Engineering, Kyoto University, Kyoto, Japan. Parentheses indicate synonymy.
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861
199
SYNONYMY OF S. CEREVISIAE AND S. UVARUM
VOL. 30, 1980
spective species (24, 25), although minor variations of physiological activities were found in
some strains.
PMR spectra of polysaccharides. The S.
cerevisiae and S. uvarum strains were divided
into type I and 11, and type I strains were subdivided into subtypes Ia and Ib on the basis of
the PMR spectra of their mannans in the H-1
region (Table 3). Subtype Ia was comprised of
three strains of S. cerevisiae and two of S. uuarum, including the type strain of S. uvarum (IF0
0615 = CBS 395). The PMR spectra of the
mannans of these strains were almost identical
to each other and were characterized by three
signals at 5.28 to 5.30, 5.17 to 5.19, and 5.08 to
5.10 ppm (Fig. 1).Subtype Ib consisted of three
strains of S. uuarum. The PMR spectra of the
mannans of this type were characterized by two
main signals at 5.28 to 5.31 and 5.08 to 5.10 ppm
and one weak signal at 5.16 to 5.20 ppm (Fig. 1).
Type I1 was comprised of three strains of S.
uvarum and the type strain of S. cerevisiae,
CBS 1171. The PMR spectra of the mannans of
these strains were characterized by two signals
at 5.28 to 5.30 and 5.08 to 5.11 ppm (Fig. 1).
Serological characteristics. (i) Slide aggludination tests for whole cells. The antigenic patterns of the s. cerevisiae and s. uvarum
strains were examined by the slide agglutination
test with related factor sera. Type Ia and Ib
strains demonstrated antigens 1, 10 (weak), and
18, and antigens 1, 10, and 18, respectively. Type
I1 strains demonstrated antigens 1and 10 (Table
4).
The serological relationships among the representative strains of each type (classified according to PMR spectral patterns) were examined by seven reciprocal absorption experiments
(Table 5). The findings from experiment 1
(Table 5) suggested that the antigenic structures
TABLE3. PMR signals from mannans of strains of S. cerevisiae and S. uvarum
Type
Signals (ppm)"
Strain
A
B
C
Ia
S. cerevisiae
S. cerevisiae
S. cerevisiae
S. uvarum
S. uvarum
6001
AJ 4001
AJ 4002
AJ 5286
I F 0 0615
5.29
5.30
5.29
5.30
5.28
5.18
5.18
5.17
5.19
5.18
5.09
5.10
5.09
5.10
5.08
Ib
S. uvarum
S. uvarum
S. uvarurn
I F 0 0565
I F 0 0751
AJ 4033
5.28
5.28
5.31
I1
S. cerevisiae
S. uvarum
S. uvarum
S. uvarum
CBS 1171
CBS 1513
I F 0 0220
I F 0 0297
5.30
5.30
5.29
5.28
M
(5.16)h
(5.16)
(5.20)
5.08
5.08
5.10
5.11
5.10
5.08
5.08
A, B, and C indicate peaks of PMR spectra.
Values in parentheses indicate weak signals.
Type
0
Ia
Type I b
81
m
m 1
M-6001
AJ-4001
AJ-4002 IFO-0615
AJ-5286
IFO- 0 5 6 5
I FO- 0 7 5 1
AJ-4033
CBS-1171 CBS-1513 IFO-0220
IFGO297
FIG. 1. PMR spectra (H-1 region) of mannans from strains of s. cerevisiae and S. uvarum. Arabic numerals
indicate chemical shifts of each signal.
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200
FUKAZAWA ET AL.
INT. J. SYST.BACTERIOL.
TABLE4. Slide agglutination of heat-killed cells of strains of S. cerevisiae and S. uvarum by factor sera
TYPe
Ia
Ib
IT
'I
Heat-killed cells of:
S. cerevisiae
S. cerevisiae
S. cereuisiae
S. uvarum
S. uvarum
M6001
AJ4001
AJ4002
AJ 5286
IF00615
S. uvarum
S. uvarum
S. uvarum
IF00565
IF00751
AJ4033
S. cerevisiae
S. uvarum
S. uuarum
S. uvarum
CBS1171
CBS1513
IF00220
IF00297
Slide agglutination with the following factor sera:"
1
+
+
+
+
+
+
+
+
+
+
+
+
4
5
6
8
9
-
-
-
-
+
+
+
w - - w - - w - - -
+
w
-
-
-
+
+
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
10
+
11
13
-
+
+
+
+
+
-
13b
-
-
-
-
-
-
-
-
-
-
- - -
18
+
+
+
+
+
+
+
+
20
-
34
-
- - - - - - - - _ _ -
-
-
-
-
-
-
-
-
-
-
-
-
+, Positive agglutination; +w, weakly positive agglutination; -, negative agglutination.
TABLE5. Absorption experiments for strains of S. cerevisiae and S. uvarum
Slide agglutination with:"
S. cerevisiae
M 6001 (Ia)
S. uvarum (la)
S. cerevisiae
CBS 1171 (11)
S. uvarum (11)
S. cerevisiae
(14
S. uuarum (Ia)
S. uuarum (11)
S. cerevisiae (11)
S. cerevisiae
(14
S. uuarum (la)
S. uuarum (Ib)
S. uvarum (Ib)
S. uuarum (Ib)
S. uvarum (Ib)
S. uvarum I F 0 0615
(la)
S. cerevisiae (Ia)
S. uvarum CBS 1513
(11)
S. cereuisiae (11)
S. uvarum (11)
S. cerevisiae (11)
S. cerevisiae (Ia)
S. uvarum (Ia)
S. uvarum IF0 0751
(Ib)
S. uvarum (Ib)
S. cerevisiae (Ia)
S. uvarum (Ia)
S. cerevisiae (11)
S. uvarum (11)
-
+, Positive agglutination; +w, weakly positive agglutination;
agglutination.
Parentheses indicate weak amount of remaining antibody.
of S. cerevisiae M 6001 (Ia) and S. uuarurn IF0
0615 (Ia) are closely related. Experiment 2
showed S. cereuisiae CBS 1171 (11) and S. uuarum CBS 1513 (11) to have identical antigenic
structures. The reactions observed in experiment 3 may be attributable to antigenic factor
18, which was reported by Tsuchiya et al. (19,
21, 22), whereas antigenic factor 10 may have
played a role in the reactions noted in experiment 4. The findings from experiment 5 sug-
+, indistinct
18
18
agglutination; -, negative
gested that a very specific part of antigen 18may
be responsible for the weak reactions noted with
type Ia strains. In experiment 7, the absorbed
sera reacted with two strains of subtype Ia and
one strain of subtype Ib, suggesting that antigenic factor 18 may be responsible for the observed positive reactions.
(ii) Quantitative precipitin tests. The
quantitative precipitin reactions of the four antisera with their homologous mannans are
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SYNONYMY OF S. CEREVISIAE A N D S. UVARUM
VOL. 30,1980
shown in Fig. 2. Precipitation was higher in the
S. cereuisiae M 6001 (Ia) and S. uvarum I F 0
0615 (Ia) systems than in the S. uuarum I F 0
0751 (Ib) system. Although no precipitate was
detected in the homologoussystem with antisera
to S. cereuisiae CBS 1171 (11)and CBS 1513 (11)
in the ordinary immunization schedule, after
long-term immunization a small amount of precipitate was formed in the homologous system
of S. cerevzszae CBS 1171. This suggests that in
the ordinary immunization schedule, S. cereuisiae M 6001 (Ia) and S. uuarum I F 0 0615 (Ia)
have strong immunogenicity and S. cereuisiae
CBS 1171 (11) and S. uvarum CBS 1513 (11)
have weak immunogenicity. The differences in
immunogenicity may depend on the mannan
side chains which act as the immunodominant
groups. Antiserum to S. cereuzsiae M 6001 (Ia)
cross-reacted with the mannans of S. uuarum
I F 0 0615 (Ia) and S. uuarum I F 0 0751 (Ib) but
not with S. uuarum CBS 1513 (11)mannan (Fig.
3). Similarly, antiserum to S. uuarum I F 0 0615
(Ia) cross-reacted with mannans of S. cerevzsiae
M 6001 (Ia) and S. uuarum I F 0 0751 (Ib) but
not with S. uuarum CBS 1513 (11) mannan.
These results suggest that the antigenic structures of S. cereuisiae M 6001 (Ia) and S. uuarum
I F 0 0615 (Ia) are almost identical, that the
antigenic structure of S. uuarum I F 0 0751 (Ib)
is closely related to the antigenic structures of
these strains, and that the antigenic structure of
S. uuarum CBS 1513 (11)is significantly different
from the antigenic structures of the three strains
mentioned above.
Gel filtration profiles of the acetolysis
oligosaccharidesof S. cereuisiae and S. UUQ-
5
10
40
20
201
00
M A N N A N ( J J 1~
FIG. 3. Precipitin curves showing cross-reactions
anti-S. cerevisiae M 6001 (la) serum with mannans
of homologous and heterologous yeast strains, Symbols: a, S. cerevisiae M 6001 (la) mannan; 0,S.
uvarum IF0 0615 (la) mannan; X, S. uvarum I F 0
0751 (Ib) mannan; A, S.uvarum CBS 1513 (II) mannan.
of
rum mannans. Figure 4 shows the gel filtration
profiles of the oligosaccharides of three serotypes of Saccharomyces species obtained by
acetolysis of mannans followed by gel filtration,
using a Bio-Gel P-2 column. The aeetolysis of S.
cereuisiae M 6001 (Ia) mannan gave five peaks
(mannose, mannobiose, mannotriose, mannotetraose, and mannopentaose, with a molar ratio,
as calculated from the fraction areas, of 2:4:5:5:
1). The acetolysis of S. uuarum I F 0 0751 mannan demonstrated four peaks (mannose, mannobiose, mannotriose, and mannotetraose, with
a molar ratio of 3:6:3:1). The acetolysis of S.
cereuisiae CBS 1171 (11)mannan gave mannose,
mannobiose, and mannotriose, with a molar ratio of 1:2:2. These results, as well as those obtained from serological studies, suggest that antigenic factor 18 of S. cereuisiae M 6001 (Ia) may
depend on mannotetraose or mannopentaose or
both.
DISCUSSION
5
10
20
40
MANNAN ( p g )
00
FIG. 2. Precipitin curves for anti-S. cerevisiae and
anti-S. uuarum sera with homologous mannans. Symbols: a, anti-S. cerevisiae M 6001 (la) serum and
homologous mannan; 0,anti-S. uvarum I F 0 0615
(la) serum and homologous mannan; X, anti-S. uvarum I F 0 0751 (Ib) serum and homologous mannan;
A, anti-S. cerevisiae CBS 11 71 (11) serum and homologous mannan.
In this study we tried to clarify the antigenic
relationships among strains of S. cereuzszae and
S. uuarum and found that the strains of the two
species could be divided into two main serotypes,
I and 11, irrespective of the species. The serological relationships correlated with the PMR spectra of the cell wall mannans. The strains of type
I characteristically demonstrated antigenic factor 18 in addition to factor 10. The strains of
type I formed mannans with closely related spectra characterized by three signals. Type I strains
could be divided further into subtypes Ia and Ib,
based on the intensity of the signals at 5.16 to
5.20 ppm (high in subtype Ia, low in subtype Ib).
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202
31
INT. J. SYST.BACTERIOL.
FUKAZAWA ET AL.
II
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k
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0
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notetraose and/or mannopentaose may be responsible for determining antigenic factor 18.
The facts that strains of S. cereuisiae and S.
uuarum can be divided into two sero- or chemovars and that strains of the two species which
belong to the same sero- or chemovar are identical strongly suggest a close phylogenetic relationship between these two species.
In comparisons of the physiological and biochemical characters of the type strains of both
species, either from their descriptions (24,25) or
from our partial examinations, it is obvious that
the two strains are very closely related to each
other. Thus, in the fermentation of 13 carbohydrates, a difference is found only in raffinose
fermentation (complete fermentation in S. uvarum and one-third fermentation in S. cereuiszae), and in the assimilation of 31 carbohydrates, a difference is found only in melibiose
assimilation, which is related to raffiose fermentation. The eight other physiological characters of the two strains are exactly the same
(Table 6).
Nakase and Komagata (12) reported that the
guanine plus cytosine contents of the deoxyribonucleic acids of S. cereuisiae, S. carlsbergensis, and S. uuarum were 38.8 to 40.2,38.5 to 39.5,
T y p e Ia
TUBES( 2.5rnl)
FIG. 4. Gel filtration profiles of the acetolysis oligosaccharides of S. cereuisiae and S. uvarum strains
on a column of Bio-Gel P-2 after elution with water.
From right to left, the carbohydrate-containingpeaks
correspond to mannose beak A), mannobiose beak
B), mannotriose beak C), mannotetraose beak D),
and mannopentaose beak E). A 490 nm, Absorbance at
490 nm.
a
Q!
0
t-
o
a
LL
0
Type I b
Z
W
Furthermore, these subtypes were characterized
by the amount of antigen 10, which was lower in
subtype Ia and higher in subtype Ib. Type I1
strains demonstrated antigen 10 but not antigen
18, and they formed mannans with very similar
spectra, which were characterized by two signals.
The relative amounts of antigenic factors 1, 10,
and 18 in type Ia, Ib, and I1 strains are shown
schematically in Fig. 5.
The individual gel filtration profiles of the
acetolysis oligosaccharides of type Ia, Ib, and I1
strains of both species were also characteristic.
Types Ia, Ib, and I1 gave fractions of five, four,
and three, oligosaccharides, respectively, suggesting that type I1 strains lack mannopentaose
and mannotetraose fragments, type Ib strains
lack mannopentaose, and type Ia strains have
both of these oligosaccharide fragments. These
results, as well as the serological characteristics
reported here, suggest the possibility that man-
c3
I-
z
a
LL
0
F
z
3
0
za
W
L
ta
W
a:
1
10
18
ANTIGENIC FACTORS
FIG. 5. Schematic diagram of relative amounts of
antigenic factors 1, 10, and 18.
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SYNONYMY OF S. CEREVISIAE AND S. UVARUM
VOL. 30, 1980
TABLE
6. Physiological and biochemical characteristics
Character
Fermentation of:
Glucose"
Galactose"
Sucrose"
Maltose"
Cellobiose
Trehalose
Lactose"
Melibiose"
Raffhose"
Melezitose
Inulin
Soluble starch
a-Meth yl-D-glucoside
Assimilation of:
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"
a-Methyl-D-glucoside"
Salicin"
DL-Lactic acid"
Succinic acid"
Citric acid"
Inositol"
Splitting of arbutin
Assimilation of nitrogen compounds:
Potassium nitrate
Ethylamine hydrochloride
Growth in vitamin-free medium
Growth on 50% (wt/wt) glucose-yeast extract agar
Growth on 60% (wt/wt) glucose-yeast extract agar
Growth at 3'7°C
Cycloheximide resistance
of
203
the type strains of S. cerevisiae and S. uvarum
S.cerevisiae
S. uvarum
CBS 1171
CBS 395
+
+OSh
+
+
+os
+
+ or -
+ or -
+ (1/3)
+os
+ (complete)
- or +
+os
-
+ or -
-, rarely +vsc
+ or +
+os
-
+ or -
+os
-
+
+os
-
+ or -
+
+ or -
- or +
-
+os
+
+ or -
- or +s"
-
- or +we
+ or + or + or -
- or ?
-
-
-
+ or + or + or + s&w or -I
-, rarely +
-
Absent
+
+ or -
+ or -, seldom +
+ or -
+vf
-, seldom +w
-
Absent
-
-
Variable
Variable
Absent
Variable
Absent
Variable
Variable
Absent
Variable
Absent
" Examined in our laboratory.
* +os, Occasionally slow reaction.
+vs, Very slow reaction.
'+s, Slow reaction.
+vw,Very weak reaction.
'+s&w, Slow and weak reaction.
+w, Weak reaction.
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204
FUKAZAWA ET AL.
INT. J. SYST.BACTERIOL.
and 39.8 mol%, respectively. In the present priority over S. uvarum, which thus becomes a
study, the guanine plus cytosine contents of S. later, subjective synonym of S. cerevisiae.
cerevisiae AJ 4001 (Ia), S. cerevisiae AJ 4002
(Ia), S. uuarum AJ 4033 (Ib), and S. uuarum
ACKNOWDGMENTS
CBS 1513 (11)deoxyribonucleic acids were 39.5,
We are very grateful to T. Tsuchiya for his critical reading
38.8, 38.5, and 39.5 mol%, respectively, when
of the manuscript and to I. Sat0 for his valuable advice. We
determined from the melting points of the de- also
thank K. Sato for performing the PMR spectral analyses.
oxyribonucleic acids. These results also suggest
a close phylogenetic relationship between the
REPRINT REQUESTS
two species.
Address reprint requests to: Dr. Y. Fukazawa, Department
Our recent discovery of a type Ib strain of S.
cerevisiae (unpublished data) confirms that of Microbiology, Meiji College of Pharmacy, 1-35-23, Nozawa,
both species contain strains of types Ia, Ib, and Setagaya-Ku, Tokyo 154, Japan.
11. The present findings regarding serovars in
LITERATURE CITED
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SYNONYMY OF S. CEREVISIAE AND S. UVARUM
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