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VIEW ARTICLE DOI: 10.1094/ASBCJ-56-0058
Rapid Methods for Detecting Saccharomyces diastaticus, a Beer Spoilage Yeast, Using the Polymerase Chain Reaction. Hiromasa Yamauchi (1), Hiroshi Yamamoto, Yuji Shibano, Noriko Amaya, and Takeshi Saeki, Institute for Fundamental Research, Technical Development Department, Suntory Ltd., 1-1-1, Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka 618, Japan; Fax: +81-75-962-9037. (1) Corresponding author. Phone: +81-75-962-9685. J. Am. Soc. Brew. Chem. 56(2):58-63, 1998. Accepted January 20, 1998.
We
have devised
rapid methods
for detecting
Saccharomyces
diastaticus,
a beer spoilage
microorganism,
using polymerase
chain reaction
(PCR). We have
designed primers
to detect S.
diastaticus
by PCR,
paying attention
to sequential
differences
between the
glucoamylase
genes of S.
diastaticus
and S. cerevisiae.
An examination
of primer reactivity
showed the forward
primer, SD-5A,
and the reverse
primer, SD-5B,
to react with
S. diastaticus
(seven strains
tested); however,
they did not
react with other
microorganisms,
including the
brewing yeast
used by our
company, two
strains of Saccharomyces
spp. 10 strains
of Brettanomyces
spp., four
strains of wild
yeast, four
strains of fungus,
and 23 strains
of various bacteria.
The DNA extracted
enzymatically
from cell numbers
as low as 10(^1)
worked successfully
as templates
for the PCR
method. Time
required to
extract DNA
from cells and
to detect S.
diastaticus
was only
approximately
5 hr. The combination
of the rapid
microbe detection
system and PCR
led to high
accuracy of
analysis. For
a test of beer
product, these
combined procedures
for the detection
of S. diastaticus
may be useful
and, even from
one cell which
becomes one
microcolony
(one to two
day old colony),
a test can be
completed in
30 hr. Keywords:
Glucoamylase
genes, Rapid
microbe detection
system (RMDS).
Hemos inventado
métodos
rápidos
para detectar
Saccharomyces
diastaticus,
un microorganismo
contaminante
de cerveza,
usando la reacción
cadena polimerasa
(PCR). Hemos
diseñado
iniciadores
para detectar
S. diastaticus
por PCR prestando
atención
a las diferencias
secuenciales
entre los genes
glucoamilasa
de S. diastaticus
y S. cerevisiae.
Una prueba de
reactividad
iniciadora mostró
el iniciador
delantero, SD-5A,
y el iniciador
opuesto, SD-5B,
para reaccionar
con S. diastaticus
(7 cepas examinadas);
sin embargo
no reaccionaron
con otros microorganismos,
incluyendo la
levadura cervecera
usada por nuestra
compañía,
2 cepas de Saccharomyces
sp., 10 cepas
de Brettanomyces
sp., 7 cepas
de levaduras
silvestres,
4 cepas de hongos,
y 23 cepas de
varias barterias.
El DNA extraído
enzimáticamente
de números
de células
tan bajos como
del orden de
10(^1) trabajaron
exitosamente
como plantillas
para el método
PCR. La combinación
del método
sistema de detección
microbiana rápida
(RMDS) y PCR
condujo a una
alta fidelidad
de análisis.
Estos procedimientos
combinados para
la detección
de S. diastaticus,
de una microcolonia
(1-2 días
colonia formada),
pueden ser completados
en 2 días.
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A subscription to the Journal of the American Society of Brewing Chemists is complimentary with membership to the ASBC. The ASBC Journal publishes scientific papers, review articles, and technical reports dealing with the chemistry and microbiology of brewing ingredients and relevant technology, as well as the analytical techniques used in the malting and brewing industry. |