Previous Article | Next Article 
J Bacteriol, June 1998, p. 3222-3226, Vol. 180, No. 12
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The Glucose Kinase of Bacillus
subtilis
Pierre
Skarlatos and
Michael K.
Dahl*
Lehrstuhl für Mikrobiologie, Institut
für Mikrobiologie, Biochemie und
Genetik, Friedrich-Alexander-Universität
Erlangen-Nürnberg, 91058 Erlangen, Germany
Received 23 February 1998/Accepted 6 April 1998
 |
ABSTRACT |
The open reading frame yqgR (now termed
glcK), which had been sequenced as part of the genome
project, encodes a glucose kinase of Bacillus subtilis. A
1.1-kb DNA fragment containing glcK complemented an
Escherichia coli strain deficient in glucose kinase
activity. Insertional mutagenesis of glcK resulted in a
complete inactivation of glucose kinase activity in crude protein
extracts, indicating that B. subtilis contains one major
glucose kinase. The glcK gene encodes a 321-residue protein
with a molecular mass of 33.5 kDa. The glucose kinase was overexpressed
as a fusion protein to a six-His affinity tag and purified to
homogeneity. The enzyme had Km values for ATP
and glucose of 0.77 and 0.24 mM, respectively, and a
Vmax of 93 µmol min
1
mg1. A B. subtilis strain deficient for
glucose kinase grew at the same rate on different carbon sources
tested, including disaccharides such as maltose, trehalose, and
sucrose.
| |
TEXT |
Glucose utilization in
Bacillus subtilis is dependent on the uptake of the sugar by
the glucose phosphoenolpyruvate-dependent phosphotransferase system
(PTS), resulting in cytoplasmic glucose-6-phosphate. One would expect
that glucose kinase is not necessary for glucose utilization. However,
unphosphorylated glucose may accumulate in the cytoplasm from
disaccharide hydrolysis, as we have previously proposed for the
trehalose system. After transport by a specific trehalose PTS (15,
27), phosphorylated trehalose-6-phosphate is hydrolyzed into
glucose-6-phosphate and glucose by the phospho-
-1,1-glucosidase TreA
(12, 15). Recently, we found additional evidence that cytoplasmic unphosphorylated glucose can be obtained by maltose or
isomaltose metabolism (28). Therefore, we have postulated the existence of a glucose kinase necessary for the phosphorylation of
the glucose released after disaccharide hydrolysis (12). Previous studies provided evidence of a glucose kinase in B. subtilis (8, 9). For Escherichia coli, as
well, the existence of internal glucose has been concluded
(21), which may originate from UDP-glucose in analogy to the
formation of free galactose from UDP-galactose (37) or by a
sugar phosphate transferase (31).
Expression of glucose kinase.
In order to investigate glucose
kinase expression, the enzymatic activity of glucose kinase was
determined in crude protein extracts from wild-type cells lysed by
sonication with a Branson sonifier (8 times, 30 s each time, 40 W)
at 4°C. Cell debris was eliminated by centrifugation, and protein
concentrations of the resulting supernatants were determined by the
method of Bradford (26). Specific glucose kinase activity
was determined in a coupled enzyme assay by the method of Seno and
Charter (30) (in a solution consisting of 50 mM Tris-HCl
[pH 7.0], 20 mM glucose, 25 mM MgCl2, 0.5 mM NADP, 1 mM
ATP, and 0.8 U of glucose-6-phosphate dehydrogenase). Glucose-6-phosphate dehydrogenase activity was assayed by monitoring the change in the optical density at 340 nm at 25°C with NADP as a
cofactor. GlcK activity was analyzed in cells in different growth
phases and in different media. As presented in Fig.
1, the activity reached a maximum at the
end of the vegetative growth period. After the cells entered the
stationary phase, the enzymatic activity slowly decreased. This latter
phenomenon could be explained by protein instability or by reduced
glcK expression when strains enter the stationary growth
phase. This result suggests that the gene is under the control of the
vegetative 
A-dependent promoter (14). An
identical expression profile has been reported for the trehalose system
(15). It has been suggested that glucose kinase contributes
to disaccharide metabolism. Therefore, similar expression profiles can
be expected.
View larger version (18K):
[in this window]
[in a new window]
|
FIG. 1.
Expression of glucose kinase activity during exponential
growth and the early stationary phase. Wild-type B. subtilis
cells were grown in Luria broth. The corresponding optical density at
600 nm of the culture is indicated. Aliquots were harvested at the
indicated times, and glucose kinase activity was determined as
described in the text and expressed in nanomoles of NADP reduced
minute1 milligram of protein1 of cell
extracts.
|
|
Glucose kinase activity has also been tested in cells grown in minimal
medium C (
22) containing several different sugars
as the
sole carbon source (20 mM each), such as glucose, fructose,
trehalose,
maltose, or sucrose. In all these cases, the glucose
kinase expression
profile resembles the profile shown in Fig.
1 (data not shown). This
result can be interpreted as a constitutive
expression of the type one
would expect for housekeeping genes.
Similar results were recently
reported for the expression of glucokinase
of
E. coli
(
21). However, we cannot exclude the possibility
that
regulation is affected on the transcriptional or posttranscriptional
level under conditions we have not tested so far.
Cloning of the glcK gene.
In a data bank search
for potential open reading frames encoding a putative glucose kinase,
one open reading frame (yqgR) from the B. subtilis genome sequencing project was identified (see Fig. 2)
(17). The corresponding region from position 2569849 to
2570922 of the B. subtilis genome (17) was cloned
by PCR amplification (23) using the following pair of
oligonucleotides: 5'-CGGTCTAGATTTTTATAAAGGCTA and
5'-CCCAAGCTTCTCATTAACACATCAA, introducing a XbaI
site 5' to glcK and a HindIII site 3' to
glcK. The 1,073-bp DNA fragment thereby obtained was ligated
behind the strong constitutive degQ36 promoter to mediate
expression in E. coli (6). The resulting plasmid,
pMD492, was able to complement an E. coli strain, UE26, (see
Table 1) deficient for glucose kinase on MacConkey agar base (Difco)
indicator plates (see Table 2) supplemented with 55 mM glucose and 1 mM
fucose (1), which is supported by a detectable glucose
kinase activity in crude protein extract (see Table 2). However, the
presence of pMD492 in E. coli UE26 does not lead to higher
enzymatic activity than that determined for the wild type.
Nucleotide sequence analysis of glcK and protein
sequence comparison.
DNA sequences were determined by the cycle
sequencing technique and ABI PRISM 310 genetic analyzer following the
recommendations of the manufacturer (Perkin-Elmer Corp., Foster City,
Calif.). We used primers 3' and 5' from the insertion (primers 1201 and 1211; New England Biolabs, Beverly, Mass.) and two internal
glcK primers 5'-CATAATCAACCACCTCAAGGGC and
5'-CCATCTGGAGACAGAAACCG. The sequences overlap each other
and covered the entire cloned DNA fragment of pMD492. The nucleotide
sequence showed that glcK is identical to
yqgR (17) with one exception, a
nucleotide change from T to G at position 2570339, which does not alter
the amino acid sequence (position numbers according to the genome
sequencing project [17]). Sequence analysis
revealed one complete open reading frame of 963 bp, starting with an
ATG initiation codon at nucleotide position 2569885 and ending with a
TAA codon at position 2570847 (Fig. 2).
The ATG codon is predicted at an appropriate distance by the putative
ribosome binding site AAGG (35).
View larger version (16K):
[in this window]
[in a new window]
|
FIG. 2.
Map of the glcK (yqgR) locus
presenting the protein and RNA coding regions at kilobase position
2570.85 on the B. subtilis genome (17). The
locations, orientations, and sizes of glcK and the flanking
open reading frames (gray arrows) are indicated by arrows. The 1.1-kb
DNA fragment containing the glcK gene whose nucleotide
sequence has been determined is presented below. The insertion of a
spc cassette between two NaeI restriction sites
leading to inactivation of glcK in strain MD186 is depicted
below the map. P1 and T1 indicate a potential promoter and terminator,
respectively. The corresponding sequences are presented below, showing
a potential A-dependent promoter with  35 and 10
regions in front of the ribosome binding site (Shine-Dalgarno sequence
[SD]) of yqgP (start codon is underlined) and a palindrome
of the suggested terminator T1.
|
|
As discussed above, the expression profile of glucose kinase indicates
that transcription is under
A promoter control.
Therefore, we looked for potential
A-dependent promoters
upstream of
glcK. The closest upstream sequence
similar to
A promoters found (marked P1 in Fig.
2), which fits the
10 region
and matches the
35 region of the
A
consensus sequence (
14) but has three changes is located 26
bp upstream of the suggested
yqgP start codon. A similar
search
was carried out for rho-independent terminators. As depicted in
Fig.
2, a putative terminator (T1), based on sequence analysis
prediction as previously described (
17), is located
downstream
of the
yqgS open reading frame. These data
obtained from sequence
analysis suggest that
glcK may be
located in an operon with two
upstream open reading frames
(
yqgP and
yqgQ) and one downstream
(
yqgS) open reading frame. Further studies must be carried
out
to prove the potential organization of a glucose kinase operon.
In the
B. subtilis genome sequencing project, the putative
yqgR gene was classified as a transcriptional repressor of
xylose-utilizing
enzymes, since it showed a homology score of 26.5%
identity for
298 amino acids to the NagC protein of
E. coli
(
17). Indeed,
the glucose kinase belongs to the so-called
ROK protein family
(regulators, open reading frames, and kinases) to
which NagC,
xylose repressors from several
bacilli, and
glucose kinases from
several organisms belong (
33). However,
the highest scores of
amino acid identities have been found for glucose
kinases (data
not shown). Furthermore, the sequence analysis revealed
no apparent
-helix-turn-
-helix motif for DNA binding in GlcK.
Glucose kinases
contain a typical AIDLGGT motif, which has been
identified as
an ATP binding site (
2). A remarkable
similarity in the nucleotide
binding core has been reported by Flaherty
et al. (
7) for yeast
hexokinase and the ATP binding regions
of 70-kDa heat shock proteins.
This N-terminal ATP-binding motif is
highly conserved in glucose
kinases and hexokinases (
29) and
is also present in GlcK of
B. subtilis. All in all, the
sequence analysis obtained strong
evidence that GlcK is the glucose
kinase.
Inactivation of glcK.
To identify the glcK
gene product and to demonstrate the physiological role of GlcK in
B. subtilis, the glcK gene was inactivated on the
chromosome. For that purpose, a 432-bp NaeI DNA fragment of
plasmid pMD492 was exchanged for a 1,158-bp
HincII-EcoRV DNA fragment of plasmid pDG1726
(13), carrying the spc gene encoding spectinomycin resistance (Table 1 and
Fig. 2). Recombinants were selected in E. coli on Luria
broth plates supplemented with ampicillin (100 µg/ml) and
spectinomycin (100 µg/ml). The construct pMD495 was verified by
restriction analysis to determine the correct resulting size of the
plasmid and the orientation of spc. This plasmid is no
longer able to reconstitute the glucose kinase phenotype in E. coli UE26 (Table 2). Inactivation of
the glucose kinase gene in B. subtilis was achieved by
transformation of linearized plasmid pMD495 in wild-type B. subtilis (Table 1) by a one-step procedure (16),
followed by selection on Luria broth supplemented with spectinomycin
(100 µg/ml). The strain was verified by Southern blot hybridization,
showing the correct insertion by homologous double-crossover
recombination (data not shown). Analysis of the resulting strain MD186
(Table 1) for glucose kinase activity showed its complete loss (Table
2). The inactivation, together with the data from complementation with
plasmid pMD492 in E. coli UE26, clearly identifies
glcK as the gene coding for glucose kinase of B. subtilis.
View this table:
[in this window]
[in a new window]
|
TABLE 2.
Glucose kinase activity in E. coli and
B. subtilis with glucose kinase or deficient for glucose
kinase activity
|
|
In a
glkA mutant of
Staphylococcus xylosus
(
glkA is the homolog of
glcK of
B. subtilis), 75% of the wild-type level of glucose
kinase activity
persists (
36). Therefore, GlkA obviously constitutes
only a
minor glucose kinase in
S. xylosus. In
B. subtilis,
glcK::
spc mutants
exhibit no significant additional glucose kinase activity,
indicating
that during vegetative growth
B. subtilis contains
only
one major glucose kinase.
Overproduction and purification of glucose kinase.
Overproduction of glucose kinase was achieved by constructing plasmid
pMD496 (Table 1). To construct pMD496, the glcK gene was
amplified by PCR, using plasmid pMD492 as a template and the following
pair of oligonucleotides: 5'-CGCGGATCCATGGACGAGATATGGTTTGCG and 5'-CCCAAGCTTCTCATTAACACATCAA, introducing a
BamHI site 5' to glcK and a
HindIII site 3' to glcK. The resulting
1,037-bp DNA fragment was ligated to the BamHI and
HindIII cloning sites of plasmid pQE9 (24),
yielding plasmid pMD496. In this plasmid glcK is fused N
terminally in frame to the six-His tag-coding region of plasmid pQE9,
leading to expression under the control of the
isopropyl-
-D-thiogalactopyranoside (IPTG)-inducible
T5 promoter in E. coli RB791 (4, 24).
The plasmid-encoded protein has a 12-residue N-terminal extension
including the affinity tag (underlined):
Met-Arg-Gly-Ser-His-His-His-His-His-His-Gly-Ser-Met... Overproduction of glucose kinase was achieved in E. coli
RB791/pMD496 after IPTG (1 mM) induction in Luria broth medium
(containing 100 µg of ampicillin per ml) for at least 1 h, as
shown by an intense protein band migrating at the expected molecular
mass of 34 kDa on a sodium dodecyl sulfate-12%
polyacrylamide gel (SDS-12%PAG) (18) (Fig.
3A). For protein purification, the cells
were harvested after 4 h of induction with IPTG. After sonication
of the cells and centrifugation, overproduced GlcK was present in the
supernatant (data not shown). GlcK was purified to homogeneity in a
Ni2+ affinity gel chromatography (24), using the
ÄKTA purifier system (Fig. 3B). During protein purification, we
analyzed the total and specific GlcK activity from each overexpression
and purification step of GlcK protein (data not shown). The enrichment of GlcK activity was calculated to be 75-fold. A 100-ml culture yielded
about 5 mg of pure protein.
View larger version (41K):
[in this window]
[in a new window]
|
FIG. 3.
Overproduction and purification of glucose kinase. (A)
Glucose kinase was overproduced as described in the text and analyzed
on an SDS-12%PAG. Aliquots were taken from uninduced cells (lane 1)
and IPTG-induced cells after 1 h (lane 2), 2 h (lane 3),
3 h (lane 4), 4 h (lane 5). The positions of molecular mass
standards (st) are indicated to the left of the gel. (B) Purification
of glucose kinase was performed by loading a Ni2+ HiTrap
chelating column with crude cell extracts from harvested cells after
4 h of IPTG induction as depicted in panel A. The elution profile
in panel B was monitored as presented by the protein absorption at 280 nm (in milliabsorption units) correlated with the column volume (1 ml).
(Insert) Four selected fractions from the elution fractions as
indicated by the gray lines are presented on an SDS-12%PAG, showing
purification of GlcK. Molecular size standards (rightmost lane) with
the indicated masses (in kilodaltons) are indicated to the right of the
inserted gel.
|
|
Properties and kinetic parameters of glucose kinase.
The
standard glucose kinase activity assay with 3 µg of purified GlcK per
ml was used to test enzyme properties and kinetic constants. Without
ATP, no significant glucose kinase activity can be observed (data not
shown). Replacement of ATP by ADP also revealed no detectable activity.
As mentioned before, GlcK contains an N-terminal, conserved typical
AIDLGGT motif, which has been identified as an ATP binding site
(2). We analyzed the ability of purified GlcK to bind ATP by
native gel electrophoresis with vertical gel slabs containing 12%
acrylamide. Prior loading, glucose kinase and 10 µCi of
[-32P]ATP diluted with unlabelled ATP to yield a final
concentration of 2.5 µM were incubated in 50 mM Tris-HCl (pH 7.5)-10
mM MgCl2 for 10 min at 20°C. The quantification of an
autoradiogram after native gel electrophoresis in a phosphorimager
showed that ATP binding depends linearly on the amount of GlcK in the
reaction mixture (data not shown). In order to distinguish whether GlcK is a hexokinase or a specific glucose kinase, we tested its ability to
phosphorylate several hexoses. Purified glucose kinase (10 µl; 0.5 mg/ml) in a buffer containing 50 mM Tris-HCl (pH 7.5) and 10 mM
MgCl2, supplemented with a 50 mM concentration of either glucose, fructose, galactose, or mannose, in the absence or presence of
50 mM ATP, was incubated at 30°C for 20 min. The same reaction mixture without protein served as a control. The resulting products were analyzed on silica-coated thin-layer chromatography plates (Merck,
Darmstadt, Germany), developed, and visualized as described previously
(21). As shown in Fig. 4, GlcK
phosphorylates glucose in the presence of ATP, whereas no
phosphorylation of fructose, galactose, or mannose could be detected
under the same conditions (data not shown). Thus, all these data
together clearly identify GlcK as an ATP-dependent glucose kinase.
Since we could demonstrate that GlcK is the B. subtilis
glucose kinase, we determined the Michaelis-Menten kinetic parameters
in a glucose kinase assay (30). GlcK had
Km values for ATP and glucose of 0.77 and 0.24 mM, respectively, and a Vmax of 93 µmol
min1 mg1.
View larger version (29K):
[in this window]
[in a new window]
|
FIG. 4.
Phosphorylation of glucose by glucose kinase GlcK.
Ten-microliter samples of each reaction mixture were spotted onto
thin-layer chromatography plates (see text). In addition to the buffer
contents, the mixtures contain glucose (lane 1), glucose-6-phosphate
(lane 2), glucose and glucose kinase (lane 3), and glucose, ATP, and
glucose kinase (lane 4).
|
|
Several parameters which influence the enzymatic activity of glucose
kinase have been tested. In order to determine the optimal
reaction
temperature, the assay has been carried out at various
temperatures
ranging from 15 to 42°C. The enzyme has an optimal
reaction
temperature of 32°C, leading to an about twofold-higher
activity
compared to the standard procedure at room temperature.
For
determination of temperature stability, glucose kinase was
preincubated
for 20 min at various temperatures ranging from 20
to 70°C, followed
by a standard glucose kinase reaction assay
at 25°C. Heat treatment
for 20 min showed half-maximal activity
at 52°C and that the enzyme
is stable up to 40°C. No detectable
enzymatic activity has been
observed at temperatures exceeding
60°C.
Metabolic function of the glucose kinase.
The B. subtilis glcK mutant MD186 (Table 1 and Fig. 2) was tested for its
ability to grow on glucose and fructose and the disaccharides maltose,
sucrose, and trehalose, respectively, as the sole carbon source in
comparison to the wild type. Surprisingly, this strain showed no
significant effect on the generation times, although it has defective
glucose kinase (data not shown). If glucose kinase
participates in disaccharide metabolism, another mechanism(s) must
be postulated to compensate for the glucose kinase-minus mutation. It
has been suggested that glucose can enter the pentose shunt via
gluconate and phosphorylation to gluconate-6-phosphate (8).
In that case, a functional glucose kinase is not necessary for
metabolism of cytoplasmic unphosphorylated glucose. This reaction could, in principle, be achieved by the glucose dehydrogenase encoded
by gdh (19). However, glucose dehydrogenase is
produced specifically during sporulation after the forespore has been
formed (10), and no enzyme activity or cross-reacting
material to antiserum against purified glucose dehydrogenase was
detected during vegetative growth (34). Therefore, the
possibility that glucose dehydrogenase plays a role in disaccharide
utilization, especially during vegetative growth, can be excluded. A
second possibility is based on the observation that glucose in its
unphosphorylated form is exported in E. coli
(25). If this were also the case in B. subtilis, glucose could reenter the cell by the glucose PTS.
The question of whether GlcK also plays a direct metabolic role in
glucose utilization cannot be answered at present. There
is evidence of
an additional glucose utilization system besides
the glucose PTS. A
glucose enzyme II (
ptsG)-minus strain of
B. subtilis is still able to grow on glucose as the sole carbon and
energy source but with 1.8 to 2.5-fold-reduced generation times,
depending on the constructs used (
11). A
ptsGHI
mutation causing
defective enzyme II
Glc, HPr, and enzyme I
completely abolishes growth on glucose (
11).
This has been
interpreted as an indication that
B. subtilis contains
an
additional glucose uptake system, which is also a PTS or is
regulated
via the PTS. The existence of two glucose transport
systems has been
described for the related organism
Bacillus licheniformis (
32): one is a PTS, and the other operates by an alternative
mechanism. If such an alternative glucose uptake system is also
present
in
B. subtilis, a glucose kinase might play a metabolic
role.
| |
ACKNOWLEDGMENTS |
We thank U. Ehmann for her interest in this work and for many
helpful suggestions and K. Oliva for editing the manuscript. This work
was carried out in the laboratories of W. Hillen, whose support is
greatly appreciated.
Financial support was obtained from the Deutsche Forschungsgemeinschaft
(Da248/2-2 and Da248/5-2), SFB473, and the Fonds der Chemischen
Industrie.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Lehrstuhl
für Mikrobiologie, FAU Erlangen-Nürnberg, Staudtstr. 5, 91058 Erlangen, Germany. Phone: 49-9131-858094. Fax:
49-9131-858082. E-mail: mdahl{at}biologie.uni-erlangen.de.
| |
REFERENCES |
| 1.
|
Angell, S.,
E. Schwarz, and M. J. Bibb.
1992.
The glucose kinase of Streptomyces coelicolor A3(2): its nucleotide sequence, transcriptional analysis and role in glucose repression.
Mol. Microbiol.
6:2833-2844[Medline].
|
| 2.
|
Arora, K. K.,
P. Shenbagamurhi,
M. Fanciulli, and P. L. Pedersen.
1990.
Glucose phosphorylation.
J. Biol. Chem.
265:5324-5328[Abstract/Free Full Text].
|
| 3.
|
Boos, W.,
U. Ehmann,
H. Forkl,
W. Klein,
M. Rimmele, and P. Postma.
1990.
Trehalose transport and metabolism in Escherichia coli.
J. Bacteriol.
172:3450-3461[Abstract/Free Full Text].
|
| 4.
|
Brent, R., and M. Ptashne.
1981.
Mechanism of action of the lexA gene product.
Proc. Natl. Acad. Sci. USA
78:4202-4208.
|
| 5.
|
Curtis, S. J., and W. Epstein.
1975.
Phosphorylation of D-glucose in Escherichia coli mutants defective in glucosephosphotransferase, mannosephosphotransferase, and glucokinase.
J. Bacteriol.
122:1189-1199[Abstract/Free Full Text].
|
| 6.
|
Dahl, M. K.,
T. Msadek,
K. Kunst, and G. Rapoport.
1991.
Mutational analysis of the Bacillus subtilis DegU regulator and its phosphorylation by the DegS protein kinase.
J. Bacteriol.
173:2539-2547[Abstract/Free Full Text].
|
| 7.
|
Flaherty, K. M.,
C. DeLuca-Flaherty, and D. B. McKay.
1990.
Three-dimensional structure of the ATPase fragment of a 70K heat-shock cognate protein.
Nature
346:623-628[Medline].
|
| 8.
|
Fortnagel, P.
1993.
Glycolysis, p. 171-180.
In
A. L. Sonenshein, J. A. Hoch, and R. Losick (ed.), Bacillus subtilis and other gram-positive bacteria; biochemistry, physiology, and molecular genetics. American Society for Microbiology, Washington, D.C.
|
| 9.
|
Freese, E.,
W. Kofart, and E. Galliers.
1970.
Commitment to sporulation and induction of glucose-phosphoenolpyruvate transferase.
Biochim. Biophys. Acta
222:265-289[Medline].
|
| 10.
|
Fujita, Y. R.,
R. Ramaley, and E. Freese.
1977.
Location and properties of glucose dehydrogenase in sporulating cells and spores of Bacillus subtilis.
J. Bacteriol.
132:282-293[Abstract/Free Full Text].
|
| 11.
|
Gonzy-Tréboul, G.,
J. H. de Waard,
M. Zagorec, and P. W. Postma.
1991.
The glucose permease of the phosphotransferase system of Bacillus subtilis: evidence for IIGlc and IIIGlc domains.
Mol. Microbiol.
5:1241-1249[Medline].
|
| 12.
|
Gotsche, S., and M. K. Dahl.
1995.
Purification and characterization of the phospho--1,1-glucosidase (TreA) of Bacillus subtilis.
J. Bacteriol.
177:2721-2726[Abstract/Free Full Text].
|
| 13.
|
Guérout-Fleury, A.-M.,
K. Shazand,
N. Frandsen, and P. Stragier.
1995.
Antibiotic-resistance cassettes for Bacillus subtilis.
Gene
167:335-336[Medline].
|
| 14.
|
Haldenwang, W. G.
1995.
The sigma factors of Bacillus subtilis.
Microbiol. Rev.
59:1-30[Abstract/Free Full Text].
|
| 15.
|
Helfert, C.,
S. Gotsche, and M. K. Dahl.
1995.
Cleavage of trehalose-phosphate in Bacillus subtilis is catalyzed by a phospho--(1,1)-glucosidase encoded by the treA gene.
Mol. Microbiol.
16:111-120[Medline].
|
| 16.
|
Kunst, F.,
T. Msadek, and G. Rapoport.
1994.
Signal transduction network controlling degradative enzyme synthesis and competence in Bacillus subtilis, p. 1-20.
In
P. J. Piggot, C. P. Moran, Jr., and P. Youngman (ed.), Regulation of bacterial differentiation. American Society for Microbiology, Washington, D.C.
|
| 17.
|
Kunst, F.,
N. Ogasawara,
I. Moszer,
A. M. Albertini,
G. Alloni,
V. Azevedo,
M. G. Bertero,
P. Bessieres,
A. Bolotin,
S. Borchert,
R. Borriss,
L. Boursier,
A. Brans,
M. Braun,
S. C. Brignell,
S. Bron,
S. Brouillet,
C. V. Bruschi,
B. Caldwell,
V. Capuano,
N. M. Carter,
S. K. Choi,
J. J. Codani,
I. F. Connerton,
A. Danchin, et al.
1997.
The complete genome sequence of the gram-positive bacterium Bacillus subtilis.
Nature
390:249-256[Medline].
|
| 18.
|
Laemmli, U. K.
1970.
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227:680-685[Medline].
|
| 19.
|
Lampel, K. A.,
B. Uratani,
G. R. Chaudhry,
R. F. Ramaley, and S. Rudikoff.
1986.
Characterization of the developmentally regulated Bacillus subtilis glucose dehydrogenase gene.
J. Bacteriol.
166:238-243[Abstract/Free Full Text].
|
| 20.
|
Messing, J.
1979.
A multipurpose cloning system based on single-stranded DNA bacteriophage M13.
Recomb. DNA Tech. Bull.
2:43-45.
|
| 21.
|
Meyer, D.,
C. Schneider-Fresenius,
R. Horlacher,
R. Peist, and W. Boos.
1997.
Molecular characterization of glucokinase from Escherichia coli K-12.
J. Bacteriol.
179:1298-1306[Abstract/Free Full Text].
|
| 22.
|
Msadek, T.,
F. Kunst,
D. Henner,
A. Klier,
G. Rapoport, and R. Dedonder.
1990.
Signal transduction pathway controlling synthesis of a class of degradative enzymes in Bacillus subtilis: expression of the regulatory genes and analysis of mutations in degS and degU.
J. Bacteriol.
172:824-834[Abstract/Free Full Text].
|
| 23.
|
Mullis, K. B., and F. A. Faloona.
1987.
Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.
Methods Enzymol.
155:335-350[Medline].
|
| 24.
|
Qiagen.
1997.
In
The QIAexpressionist, 3rd ed.
Qiagen Inc., Hilden, Germany.
|
| 25.
|
Rimmele, M., and W. Boos.
1994.
Trehalose-6-phosphate hydrolase of Escherichia coli.
J. Bacteriol.
176:5654-5664[Abstract/Free Full Text].
|
| 26.
|
Rodriguez, R. L., and R. C. Tait.
1983.
In
Recombinant DNA techniques: an introduction.
Addison-Wesley Publishing Co., Reading, Mass.
|
| 27.
|
Schöck, F., and M. K. Dahl.
1996.
Analysis of DNA flanking the treA gene of Bacillus subtilis reveals genes encoding a putative specific enzyme IITre and a potential regulator of the trehalose operon.
Gene
175:59-63[Medline].
|
| 28.
|
Schönert, S.,
T. Buder, and M. K. Dahl.
1998.
Identification and enzymatic characterization of the maltose-inducible -glucosidase MalL (sucrase-isomaltase-maltase) of Bacillus subtilis.
J. Bacteriol.
180:2574-2578[Abstract/Free Full Text].
|
| 29.
|
Schwab, D. A., and J. E. Wilson.
1991.
Complete amino acid sequence of the type III isozyme of rat hexokinase, deduced from the cloned cDNA.
Arch. Biochem. Biophys.
285:365-370[Medline].
|
| 30.
|
Seno, E. T., and K. F. Charter.
1983.
Glycerol catabolic enzymes and their regulation in wild type and mutant strains of Streptomyces coelicolor A3(2).
J. Gen. Microbiol.
129:1403-1413[Abstract/Free Full Text].
|
| 31.
|
Stevens-Clark, J. S.,
M. C. Theisen,
K. A. Conklin, and R. A. Smith.
1968.
Phosphoramidates. VI. Purification and characterization of a phosphoryl transfer enzyme from Escherichia coli.
J. Biol. Chem.
243:4468-4473[Abstract/Free Full Text].
|
| 32.
|
Tangney, M.,
F. G. Priest, and W. J. Mitchell.
1993.
Two glucose transport systems in Bacillus licheniformis.
J. Bacteriol.
175:2137-2142[Abstract/Free Full Text].
|
| 33.
|
Titgemeyer, F.,
J. Reizer,
A. Reizer, and J. M. H. Saier.
1994.
Evolutionary relationships between sugar kinases and transcriptional repressors in bacteria.
Microbiology
140:2349-2354[Abstract/Free Full Text].
|
| 34.
|
Vasantha, N.,
B. Uratani,
R. Ramalay, and E. Freese.
1983.
Isolation of a developmental gene of Bacillus subtilis and its expression in Escherichia coli.
Proc. Natl. Acad. Sci. USA
80:785-789[Abstract/Free Full Text].
|
| 35.
|
Vellanoweth, R. L.
1993.
Translation and its regulation, p. 699-711.
In
A. L. Sonenshein, J. A. Hoch, and R. Losick (ed.), Bacillus subtilis and other gram-positive bacteria: biochemistry, physiology, and molecular genetics. American Society for Microbiology, Washington, D.C.
|
| 36.
|
Wagner, E.,
S. Marcandier,
O. Egeter,
J. Deutscher,
F. Götz, and R. Brückner.
1995.
Glucose kinase-dependent catabolite repression in Staphylococcus xylosus.
J. Bacteriol.
177:6144-6152[Abstract/Free Full Text].
|
| 37.
|
Wu, H. C. P.
1967.
Role of galactose transport system in the establishment of endogenous induction of the galactose operon in Escherichia coli.
J. Mol. Biol.
24:213-223[Medline].
|
J Bacteriol, June 1998, p. 3222-3226, Vol. 180, No. 12
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Seibold, G. M., Wurst, M., Eikmanns, B. J.
(2009). Roles of maltodextrin and glycogen phosphorylases in maltose utilization and glycogen metabolism in Corynebacterium glutamicum. Microbiology
155: 347-358
[Abstract]
[Full Text]
-
Han, B., Liu, H., Hu, X., Cai, Y., Zheng, D., Yuan, Z.
(2007). Molecular Characterization of a Glucokinase with Broad Hexose Specificity from Bacillus sphaericus Strain C3-41. Appl. Environ. Microbiol.
73: 3581-3586
[Abstract]
[Full Text]
-
Thomaides, H. B., Davison, E. J., Burston, L., Johnson, H., Brown, D. R., Hunt, A. C., Errington, J., Czaplewski, L.
(2007). Essential Bacterial Functions Encoded by Gene Pairs. J. Bacteriol.
189: 591-602
[Abstract]
[Full Text]
-
Schonert, S., Seitz, S., Krafft, H., Feuerbaum, E.-A., Andernach, I., Witz, G., Dahl, M. K.
(2006). Maltose and Maltodextrin Utilization by Bacillus subtilis. J. Bacteriol.
188: 3911-3922
[Abstract]
[Full Text]
-
Brigham, C. J., Malamy, M. H.
(2005). Characterization of the RokA and HexA Broad-Substrate-Specificity Hexokinases from Bacteroides fragilis and Their Role in Hexose and N-Acetylglucosamine Utilization. J. Bacteriol.
187: 890-901
[Abstract]
[Full Text]
-
Lunin, V. V., Li, Y., Schrag, J. D., Iannuzzi, P., Cygler, M., Matte, A.
(2004). Crystal Structures of Escherichia coli ATP-Dependent Glucokinase and Its Complex with Glucose. J. Bacteriol.
186: 6915-6927
[Abstract]
[Full Text]
-
Mukai, T., Kawai, S., Matsukawa, H., Matuo, Y., Murata, K.
(2003). Characterization and Molecular Cloning of a Novel Enzyme, Inorganic Polyphosphate/ATP-Glucomannokinase, of Arthrobacter sp. Strain KM. Appl. Environ. Microbiol.
69: 3849-3857
[Abstract]
[Full Text]
-
Dorr, C., Zaparty, M., Tjaden, B., Brinkmann, H., Siebers, B.
(2003). The Hexokinase of the Hyperthermophile Thermoproteus tenax: ATP-DEPENDENT HEXOKINASES AND ADP-DEPENDENT GLUCOKINASES, TWO ALTERNATIVES FOR GLUCOSE PHOSPHORYLATION IN ARCHAEA. J. Biol. Chem.
278: 18744-18753
[Abstract]
[Full Text]
-
Hansen, T., Reichstein, B., Schmid, R., Schonheit, P.
(2002). The First Archaeal ATP-Dependent Glucokinase, from the Hyperthermophilic Crenarchaeon Aeropyrum pernix, Represents a Monomeric, Extremely Thermophilic ROK Glucokinase with Broad Hexose Specificity. J. Bacteriol.
184: 5955-5965
[Abstract]
[Full Text]
-
Lee, J., Blaschek, H. P.
(2001). Glucose Uptake in Clostridium beijerinckii NCIMB 8052 and the Solvent-Hyperproducing Mutant BA101. Appl. Environ. Microbiol.
67: 5025-5031
[Abstract]
[Full Text]
-
Zalieckas, J. M., Wray, L. V. Jr., Fisher, S. H.
(1999). trans-Acting Factors Affecting Carbon Catabolite Repression of the hut Operon in Bacillus subtilis. J. Bacteriol.
181: 2883-2888
[Abstract]
[Full Text]
-
Zalieckas, J. M., Wray, L. V. Jr., Fisher, S. H.
(1998). Expression of the Bacillus subtilis acsA Gene: Position and Sequence Context Affect cre-Mediated Carbon Catabolite Repression. J. Bacteriol.
180: 6649-6654
[Abstract]
[Full Text]
Top
Abstract
Text
