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Journal of Bacteriology, June 1999, p. 3705-3709, Vol. 181, No. 12
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Modes of Action of Five Different Endopectate
Lyases from Erwinia chrysanthemi 3937
Caroline
Roy,1,2
Harry
Kester,1
Jaap
Visser,1
Vladimir
Shevchik,2
Nicole
Hugouvieux-Cotte-Pattat,2
Jeanine
Robert-Baudouy,2 and
Jacques
Benen1,*
Molecular Genetics of Industrial
Micro-organisms, Wageningen Agricultural University, 6703 HA
Wageningen, The Netherlands,1 and
Laboratoire de Génétique Moléculaire des
Micro-organismes et des Interactions Cellulaires, UMR-CNRS 5577, INSA, 69 621 Villeurbanne Cedex, France2
Received 23 November 1998/Accepted 17 April 1999
 |
ABSTRACT |
Five endopectate lyases from the phytopathogenic bacterium
Erwinia chrysanthemi, PelA, PelB, PelD, PelI, and PelL,
were analyzed with respect to their modes of action on polymeric and
oligomeric substrates (degree of polymerization, 2 to 8). On
polygalacturonate, PelB showed higher reaction rates than PelD, PelI,
and PelA, whereas the reaction rates for PelL were extremely low. The
product progression during polygalacturonate cleavage showed a typical
depolymerization profile for each enzyme and demonstrated their
endolytic character. PelA, PelI, and PelL released oligogalacturonates
of different sizes, whereas PelD and PelB released mostly unsaturated
dimer and unsaturated trimer, respectively. Upon prolonged incubation, all enzymes degraded the primary products further, to unsaturated dimer
and trimer, except for PelL, which degraded the primary products to
unsaturated tetramer and pentamer in addition to unsaturated dimer and
trimer. The bond cleavage frequencies on oligogalacturonates revealed
differences in the modes of action of these enzymes that were
commensurate with the product progression profiles. The preferential products formed from the oligogalacturonates were unsaturated dimer for
PelD, unsaturated trimer for PelB, and unsaturated tetramer for PelI
and PelL. For PelA, preferential products were dependent on the sizes
of the oligogalacturonates. Whereas PelB and PelD displayed their
highest activities on hexagalacturonate and tetragalacturonate, respectively, PelA, PelI, and PelL were most active on the octamer, the
largest substrate used. The bond cleavage frequencies and reaction
rates were used to estimate the number of subsites of each enzyme.
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INTRODUCTION |
Erwinia chrysanthemi is a
plant pathogenic enterobacterium which causes soft-rot disease. This
bacterium secretes various enzymes that permit the degradation of
pectin, which is present in the middle lamella and primary plant cell
walls. Pectin is a complex carbohydrate consisting of smooth
homogalacturonan regions, where the backbone is partially
methylesterified and/or acetylated (4), and of the so-called
hairy regions, consisting of alternating galacturonate-rhamnose
stretches. To the rhamnose residues, side chains like arabinans or
galactans can be attached (17). To date, pectin
methylesterase, pectin acetylesterase, pectate lyase, pectin lyase, and
polygalacturonase activities have been detected in E. chrysanthemi (1, 6, 19). Among those activities, the
pectate lyases appear to be the most abundant, and they have been shown
to play an important role in soft-rot disease (2). The
pectate lyases cleave internal glycosidic bonds in polygalacturonate by
-elimination, releasing both saturated and unsaturated
oligogalacturonates. The pectate lyase activity of E. chrysanthemi results from the combined action of numerous pectate
lyases, including five major enzymes, PelA, PelB, PelC, PelD, and PelE
(23), and five minor enzymes, PelX (3, 21), PelL
(10, 18), PelZ (14), PelI (20), and
PelW (22). All pectate lyases are extracellular and have an
endo mode of action, except for PelW and PelX, which are localized in
the cytoplasm and the periplasm, respectively, and have an exolytic
activity (21, 22).
With three-dimensional (3D) structures now available for three pectate
lyases, namely, Plc and Ple from E. chrysanthemi EC16 (9, 24, 25) and Pel from Bacillus subtilis
(13), we have addressed the mode of action of five pectate
lyases to understand their specificity as determined by the nature and
number of subsites. PelA, PelB, and PelD represent the acidic, neutral,
and basic pectate lyases from family 1, respectively (5),
and PelI and PelL are representatives of families 3 and 4, respectively
(20). The enzymes were studied with respect to bond cleavage
frequencies and reaction rates on oligogalacturonates with degrees of
polymerization (n) of 2 to 8 and with respect to the
reaction rates and the product progression on polygalacturonate. This
provides a quantitative basis for their different modes of action and
allows an estimation of the number of subsites.
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MATERIALS AND METHODS |
Abbreviations.
The abbreviations used are as follows: Pel,
pectate lyase; (GalpA)n,
oligogalacturonate with indicated degree of polymerization
(n); u(GalpA)n,
4,5-unsaturated oligogalacturonate with indicated degree of
polymerization (n); HPAEC, high-performance anion-exchange
chromatography; PAD, pulsed amperometric detection.
Overproduction of the pectate lyases.
The bacterial strains,
plasmids, and techniques used to overproduce and purify the five
pectate lyases examined in the present study were described previously
(18, 20, 23). Protein concentrations were determined by the
Bio-Rad protein assay with bovine serum albumin as a standard.
Substrates.
The oligogalacturonates of defined length
(n = 2 to 8) were prepared as described previously
(7). Polygalacturonate (degree of polymerization, 150 [8]) was obtained from U.S. Biochemical Corp.
(Cleveland, Ohio).
Enzymatic assays.
Activities on polygalacturonate and
oligogalacturonates were determined spectrophotometrically by measuring
the change in absorbance at 235 nm due to the formation of
4,5-unsaturated products (
235 = 4,600 M
1 · cm1 [11]). All
assays were conducted at 37°C in 0.5 ml with 2.5 g of
polygalacturonate liter1 or 0.5 mM oligogalacturonate
(n = 2 to 8) in 20 mM buffer-1 mM CaCl2.
The buffers used were Tris-HCl (pH 8.0) for PelA, PelD, and PelL and
2-amino-methyl-propanol-HCl (pH 9.0) for PelB and PelI. These buffers
were chosen after determination of the pH optima. One unit of pectate
lyase activity is the amount of enzyme which liberates 1 µmol of
product per min.
Mode of action.
For the determination of the bond cleavage
frequencies on oligogalacturonates (n = 2 to 8) and for
the analysis of the products released from polygalacturonate, enzyme
reactions were performed under conditions identical to those described
for the enzymatic assays: 2.5 g of polygalacturonate
liter1 or 0.5 mM oligogalacturonate (n = 2 to 8), 1 mM CaCl2, and 20 mM Tris-HCl (pH 8.0) (for
PelA, PelD, and PelL) or 2-amino-methyl-propanol-HCl (pH 9.0) (for
PelB and PelI).
To determine the bond cleavage frequencies, enzyme amounts were chosen
to minimize the risk of secondary cleavage product formation. Reactions
were stopped by lowering the pH to 3.5 to 4 by the addition of 0.2 volume of 1.2% (vol/vol) acetic acid. Reaction products were analyzed
by HPAEC with PAD (Dionex Inc., Sunnyville, Calif.) and UV detection at
235 nm (Pye-Unicam LC-UV detector). The elution conditions used were
the same as those outlined previously (12). The saturated
products were quantitated from the PAD signal and the PAD response from
a calibration mixture of oligogalacturonates (n = 1 to
8) at a concentration of 0.1 mM each. The unsaturated products formed
were estimated from the peak areas of the UV profile. From the combined
data, the bond cleavage frequencies were calculated.
The product formation during polygalacturonate cleavage was studied for
up to 24 h. Normalized amounts of enzyme were added
(1.80 U/ml for
PelA, 2.13 U/ml for PelB, 1.87 U/ml for PelD, 2.08
U/ml for PelI, and
1.90 U/ml for PelL). At timed intervals, 50-µl
samples were withdrawn
and reactions were stopped by lowering
the pH to 3.5 to 4 by the
addition of 0.2 volume of 1.2% (vol/vol)
acetic acid. Reaction
products were analyzed by HPAEC as described
above. The unsaturated
products formed were quantitated from the
UV profiles and a standard
containing unsaturated dimer at 0.2
mM. For each enzyme, the percentage
of substrate conversion after
24 h was estimated from the combined
mass of the unsaturated products
formed as quantified by
HPAEC.
3D structure modelling.
In order to model the 3D structures
of PelB (accession no. X67475) and PelD (accession no. AJ132101) based
on the E. chrysanthemi EC16 Plc (PDB accession no. 2PEC) and
Ple (PDB accession no. 1PCL) structures, respectively, the Modeler
routine of the Homology module as provided by the program Insight II
(version 97.0; Molecular Simulations Inc., San Diego, Calif.) was used. The quality of the models was estimated from Ramachandran plots.
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RESULTS AND DISCUSSION |
Reaction rates and product progression on polygalacturonate.
The five enzymes studied showed very different specific activities on
polygalacturonate (Table 1). PelB and
PelD were the most active, whereas PelI and PelA showed less activity.
The activity of PelL was very low.
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TABLE 1.
Specific activities of E. chrysanthemi 3937 endopectate lyases at their pH optima with polygalacturonate as
the substratea
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The results of the product progression data for each enzyme are
presented in Fig.
1. For clarity, only
the data for the period
up to 4 h of incubation are shown. Only
minor further changes
were observed after 4 h. These involved
mainly a small increase
in u(Gal
pA)
2, except in
the case of PelL, which revealed a further
increase in
u(Gal
pA)
2 and u(Gal
pA)
3
at the expense of u(Gal
pA)
5 and
u(Gal
pA)
6. After 24 h, 70 to 78% of the
polygalacturonate
initially present, depending on the enzyme, was
converted (data
not shown). For all five enzymes, the data showed the
initial
formation of oligogalacturonates larger than
u(Gal
pA)
5, which
were gradually converted into
smaller products, compatible with
an endolytic mode of attack of the
polymer substrate. The depolymerization
profiles, which are a result of
the number of subsites, the affinity
of the subsite(s) for the
substrate, and the intrinsic reaction
rate, are different for all
enzymes. For PelB and PelD, one major
product accumulated quickly,
namely, u(Gal
pA)
3 and
u(Gal
pA)
2,
respectively, indicating a strong
preference for the substrate
to bind in the particular mode resulting
in those products. Furthermore,
for PelB, a strong accumulation of
u(Gal
pA)
2 was also observed,
although with a
lag. This suggests that smaller products bind
in a different preferred
productive mode as compared to the larger
products. For PelA, PelI, and
PelL, less pronounced favored products
were formed. PelA converted the
initial larger products formed
into u(Gal
pA)
2
and u(Gal
pA)
3. For PelI, a prominent transient
increase in u(Gal
pA)
4, which was degraded into
u(Gal
pA)
2 upon
further incubation was recorded.
PelL also showed accumulation
of u(Gal
pA)
4 and
u(Gal
pA)
5, but u(Gal
pA)
4
was not further cleaved.
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FIG. 1.
Unsaturated reaction products formed upon incubation of
polygalacturonate with the E. chrysanthemi 3937 pectate
lyases PelA (A), PelB (B), PelD (C), PelI (D), and PelL (E). Conditions
are described in Materials and Methods. The following concentrations of
enzymes were added: 1.80 U · ml1 for PelA, 2.13 U · ml1 for PelB, 1.87 U · ml1 for PelD, 2.08 U · ml1 for PelI,
and 1.90 U · ml1 for PelL. uGn,
u(GalpA)n. Symbols:  ,
u(GalpA)2;  ,
u(GalpA)3;  ,
u(GalpA)4;  ,
u(GalpA)5;  ,
u(GalpA)6;  ,
u(GalpA)7-10.
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Previously, product progression profiles were reported for Pla, Plb,
Plc, and Ple from
E. chrysanthemi EC16 (
15). Pla,
Plc,
and Ple are most closely related to PelA, PelB, and PelD of
E. chrysanthemi 3937, respectively, with 29 amino acid
substitutions
for Pla, 27 amino acid substitutions for Plc, and 22 amino acid
substitutions and 7 amino acid insertions for Ple.
Surprisingly,
despite these mutations, the product progression profiles
for
PelA and PelD closely resemble those for Pla and Ple
(
15). However,
for PelB, a strong accumulation of
u(Gal
pA)
2, which was in contrast
to the profile
for Plc, was observed. The origin of these similarities
and differences
will be discussed
below.
Reaction rates and bond cleavage frequencies on oligogalacturonates
(n = 2 to 8).
In order to understand the origin
of the differences between the individual enzymes in product
progression on polygalacturonate and their mode of action, the turnover
rates of oligogalacturonates (n = 2 to 8) and
corresponding bond cleavage frequencies for each enzyme were studied.
None of these endo-acting enzymes showed degradation of
(Gal
pA)
2, which is also reflected in the product
progression profiles
where u(Gal
pA)
2 accumulated
as the smallest product. Another major
product observed in the product
progression profile was u(Gal
pA)
3.
The fact that
this product accumulated must be attributed to the
very low conversion
rates of this substrate by all five enzymes,
as revealed in studies
using unsaturated oligomers as substrates
(data not shown). This is
also corroborated by the observed low
reaction rates on
(Gal
pA)
3 for all five enzymes (Table
2). Thus,
the enzymes require substrates
with an
n of >3 for effective catalysis.
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TABLE 2.
Bond cleavage frequencies and specific activities of the
E. chrysanthemi 3937 endopectate lyases PelA, PelI, PelL,
PelD, and PelB acting on oligogalacturonates of
defined lengthsa
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For PelA, two preferred cleavage positions were observed depending on
the size of the oligogalacturonate. Exclusive cleavage
at the second
bond from the reducing end for (Gal
pA)
3 and
(Gal
pA)
4 and preferential cleavage at this bond
for (Gal
pA)
5 and (Gal
pA)
6 were found, whereas (Gal
pA)
7 and
(Gal
pA)
8 were preferentially
cleaved at the
second bond from the nonreducing end. This shift
of the preferred bond
cleaved, together with the observation that
the penultimate bond from
both ends was preferred over more central
bonds for oligogalacturonates
with an
n of >5, strongly indicates
that the number of
subsites with the highest affinity for homogalacturonan
is four,
stretching from positions
2 to +2. This is further substantiated
by
the lower cleavage rates for (Gal
pA)
5 and
(Gal
pA)
6 relative
to that for
(Gal
pA)
4. The increased reaction rate for
(Gal
pA)
7 and (Gal
pA)
8
indicates that the actual number of subsites exceeds
four.
Despite a 100-fold difference in reaction rates between PelI and PelL,
both enzymes showed their highest activities on the
longer
oligogalacturonates (
n 
6) and displayed comparable
bond
cleavage frequencies, with a preference for the fourth glycosidic
linkage from the reducing end. Subsequent conversion of the smaller
oligomers that were formed proceeded for both enzymes at a much
lower
rate, particularly for PelL, and with different end products.
For PelI,
u(Gal
pA)
4 was still a good substrate, explaining
the
decrease of this product in the progression profiles. Due to the
relatively much lower activity of PelL on
(Gal
pA)
5 and (Gal
pA)
4,
these substrates are not further converted as long as
oligogalacturonates
with a degree of polymerization of >5 are present.
For the smaller
oligogalacturonates, the preferred cleavage was
observed at the
nonreducing end for PelI whereas PelL preferred the
second glycosidic
bond from the reducing end. Since the reaction rates
for PelI
and PelL increased with oligogalacturonates with an
n up to 7
with increasing preference for the fourth
glycosidic linkage from
the reducing end, it can be concluded that for
both enzymes the
minimum number of subsites is seven, stretching from
positions
3 to +4. It should be pointed out that in view of the very
low
activity of PelL, but its pronounced macerating activity
(
20),
homogalacturonan may not be the natural
substrate.
PelD was the most prominent enzyme with respect to the preferred bond
cleaved. For oligogalacturonates with an
n of 6, hardly
any
products other than u(Gal
pA)
2 were detected, and
for those
with an
n of 7 or 8, the second bond from the
reducing end was
still strongly preferred. Thus, the bond cleavage
frequencies
explain the product progression well. The strong preference
of
PelD for the second glycosidic linkage from the reducing end and
the
highest activity on (Gal
pA)
4 strongly suggest
that the array
of four subsites stretches from positions
2 to +2.
For PelB, cleavage of oligogalacturonates with an
n of >4
occurred predominantly at the third glycosidic bond from the reducing
end whereas u(Gal
pA)
2 was formed mainly from
(Gal
pA)
4 and (Gal
pA)
3.
The highest reaction rate was observed on
(Gal
pA)
6. In combination
with the predominant
cleavage of oligogalacturonates with an
n of >4 at the
third linkage from the reducing end, this indicates
the presence of six
subsites extending from positions
3 to +3.
As mentioned above, the product progression for PelB differed from that
of Plc, the
E. chrysanthemi EC16 counterpart, mainly
with
respect to u(Gal
pA)
2 formation. A comparison of
the data
for PelB with those obtained for Plc under slightly different
conditions (
16) reveals only small changes with respect to
bond
cleavage frequencies and even a stronger preference for Plc to
form u(Gal
pA)
2 from
(Gal
pA)
4 (75%). However, the rate at which
(Gal
pA)
4 is cleaved by Plc relative to that of
the larger oligogalacturonates
is lower than the same cleavage ratio
for PelB
[(Gal
pA)
4/(Gal
pA)
6 cleavage ratios for Plc and PelB are 1 to 4.6 and 1 to 1.7, respectively].
This indeed can account for the observed difference in
product
progression.
Since PelB and Plc differ with respect to only 27 of 353 amino acids,
the PelB 3D structure was modeled by using the Plc 3D
structure
(
16). Most amino acid substitutions took place at
a site
that was remote from the substrate binding cleft (
16)
in the
N-terminal part of the enzyme that forms an
-helix capping
the
-helix and at the surface of the enzyme. Those mutations
are,
therefore, probably not involved in enzyme specificity. However,
three
amino acid substitutions (G161N, A167S, and S239G; the last
residues
refer to those of Plc) were found in the substrate binding
cleft. At
least one of the substitutions, S239G, may involve (in)direct
contact
with the substrate due to the change of the functionality
of the side
chain. In the modeled PelB structure, the G161N mutation
results in a
changed conformation of D160. In the Plc-substrate
structure, D160 and
D162 coordinate a Ca
2+ ion that is in contact with the
substrate (
16). The A167S mutation
most probably does not
affect the enzymatic properties since its
position below the
Ca
2+ ion present in both the substrate-free enzyme and the
enzyme-substrate
complex is not likely to influence the
Ca
2+ ion that is coordinated by the four conserved acidic
residues
D129, D131, E166, and D170. Thus, the observed changes in the
modeled 3D structure of PelB, notably G161N and S239G, may well
account
for the different product progression profiles of PelB
and
Plc.
The limited number of amino acid differences observed for PelD and Ple
also allowed the modeling of the PelD 3D structure
based on the Ple 3D
structure (
9). None of the amino acid substitutions
was
found to occur in the substrate binding cleft as inferred
from the
Plc-substrate complex (
16). The similar product progression
profiles for PelD and Ple and those differing for PelB and Plc
indicate
that the naturally occurring mutations do not change
the catalytic
properties unless they occur in the substrate binding
cleft. From this,
it logically follows that the amino acid differences
between PelA and
Pla are most likely in positions remote from
the substrate binding
cleft.
For all of the enzymes studied here, the bond cleavage frequencies and
substrate reaction rates on oligogalacturonates (
n = 2
to 8) easily explain the product progression on the model
substrate
polygalacturonate. Still puzzling are the much higher
activities of
PelB and PelI on oligogalacturonates relative to
those on
polygalacturonate. A plausible explanation would be that
the polymer
substrate obviously extends beyond the array of subsites
and that this
hampers its effective
binding.
In physiological terms, PelD, PelB, and PelI, being the most active
enzymes and generating u(Gal
pA)
2,
u(Gal
pA)
3, and u(Gal
pA)
4,
respectively, might serve as the major substrate supply for the
bacterium. Both u(Gal
pA)
2 and
u(Gal
pA)
3 may be internalized directly
(
6). PelX might cleave u(Gal
pA)
4 into
u(Gal
pA)
2 (
22).
It has been shown that PelD and PelI are very active in tissue
maceration, that PelB and PelL are moderately active in this
respect,
and that PelA is virtually devoid of this activity (
1,
10,
20). So far, we have not been able to establish a relationship
between specific activity, mode of action, and macerating properties
of
the enzymes. Further elucidation of the substrate specificity
of the
pectate lyases must await the isolation of well-defined
complex
substrates and the elucidation of 3D structures of enzyme-substrate
complexes.
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ACKNOWLEDGMENT |
This work was supported by E.C. grant AIR2-CT941345 to J.V. and
J.R.-B.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Molecular
Genetics of Industrial Micro-organisms, Wageningen Agricultural
University, Dreijenlaan 2, 6703 HA Wageningen, The Netherlands. Phone:
31 317484439. Fax: 31 317484011. E-mail:
jac.benen{at}algemeen.mgim.wau.nl.
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REFERENCES |
| 1.
|
Barras, F.,
F. Van Gijsegem, and A. K. Chatterjee.
1994.
Extracellular enzymes and pathogenesis of soft-rot Erwinia.
Annu. Rev. Phytopathol.
32:201-234.
|
| 2.
|
Boccara, M.,
A. Diolez,
M. Rouve, and A. Kotoujansky.
1988.
The role of individual pectate lyases of Erwinia chrysanthemi strain 3937 in pathogenicity on Saintpaulia plants.
Physiol. Mol. Plant Pathol.
33:95-104.
|
| 3.
|
Brooks, A. D.,
S. Y. He,
S. Gold,
N. T. Keen,
A. Collmer, and S. W. Hutcheson.
1990.
Molecular cloning of the structural gene for exopolygalacturonate lyase from Erwinia chrysanthemi EC16 and characterization of the enzyme product.
J. Bacteriol.
172:6950-6958[Abstract/Free Full Text].
|
| 4.
|
De Vries, J. A.,
M. Hansen,
J. Søderberg,
P. E. Glahn, and J. K. Pedersen.
1986.
Distribution of methoxyl groups in pectins.
Carbohydr. Polym.
6:165-176.
|
| 5.
|
Henrissat, B.,
S. E. Heffron,
M. D. Yoder,
S. E. Lietzke, and F. Jurnak.
1995.
Functional implications of structure-based sequence alignment of proteins in the extracellular pectate lyase superfamily.
Plant Physiol.
107:963-976[Abstract].
|
| 6.
|
Hugouvieux-Cotte-Pattat, N.,
G. Condemine,
W. Nasser, and S. Reverchon.
1996.
Regulation of pectinolysis in Erwinia chrysanthemi.
Annu. Rev. Microbiol.
50:213-257[Medline].
|
| 7.
|
Kester, H. C. M., and J. Visser.
1990.
Purification and characterization of polygalacturonases produced by the hyphal fungus Aspergillus niger.
Biotechnol. Appl. Biochem.
12:150-160[Medline].
|
| 8.
|
Kester, H. C. M.,
M. A. Kusters-van Someren,
Y. Müller, and J. Visser.
1996.
Primary structure and characterization of an exopolygalacturonase from Aspergillus tubingensis.
Eur. J. Biochem.
240:738-746[Medline].
|
| 9.
|
Lietzke, S. E.,
R. D. Scavetta,
M. D. Yoder, and F. Jurnak.
1996.
The refined three-dimensional structure of pectate lyase E from Erwinia chrysanthemi at 2.2 Å resolution.
Plant Physiol.
111:73-92[Abstract].
|
| 10.
|
Lojkowska, E.,
C. Masclaux,
M. Boccara,
J. Robert-Baudouy, and N. Hugouvieux-Cotte-Pattat.
1995.
Characterization of the pelL gene encoding a novel pectate lyase of Erwinia chrysanthemi 3937.
Mol. Microbiol.
16:1183-1195[Medline].
|
| 11.
|
MacMillan, G. P., and R. H. Vaughn.
1964.
Purification and properties of a polygalacturonate-trans-eliminase produced by Clostridium multifermentans.
Biochemistry
3:564-572.
|
| 12.
|
Pa enicová, L.,
J. A. E. Benen,
H. C. M. Kester, and J. Visser.
1998.
pgaE encodes a fourth member of the endopolygalacturonase gene family from Aspergillus niger.
Eur. J. Biochem.
251:72-80[Medline].
|
| 13.
|
Pickersgill, R.,
J. Jenkins,
G. Harris,
W. Nasser, and J. Robert-Baudouy.
1994.
The structure of Bacillus subtilis pectate lyase in complex with calcium.
Nat. Struct. Biol.
1:717-723[Medline].
|
| 14.
|
Pissavin, C.,
J. Robert-Baudouy, and N. Hugouvieux-Cotte-Pattat.
1996.
Regulation of pelZ, a gene of the pelB-pelC cluster encoding a new pectate lyase of Erwinia chrysanthemi 3937.
J. Bacteriol.
178:7187-7196[Abstract/Free Full Text].
|
| 15.
|
Preston, J. F.,
J. D. Rice,
L. O. Ingram, and N. T. Keen.
1992.
Differential depolymerization mechanisms of pectate lyases secreted by Erwinia chrysanthemi EC16.
J. Bacteriol.
174:2039-2042[Abstract/Free Full Text].
|
| 16.
| Scavetta, R. D., S. R. Herron, A. T. Hotchkiss, N. Kita, N. T. Keen, J. A. E. Benen, H. C. M. Kester, J. Visser, and F. Jurnak. Structure of plant
cell wall fragment complexed to PelC. Plant Cell, in press.
|
| 17.
|
Schols, H. A.,
M. A. Posthumus, and A. G. J. Voragen.
1990.
Structural features of hairy regions of pectins isolated from apple juice produced by the liquefaction process.
Carbohydr. Res.
206:117-130.
|
| 18.
|
Shevchik, V. E.,
M. Scott,
O. Mayans, and J. Jenkins.
1998.
Crystallization and preliminary X-ray analysis of a member of a new family of pectate lyases, PelL from Erwinia chrysanthemi.
Acta Crystallogr. Sect. D.
54:419-422[Medline].
|
| 19.
|
Shevchik, V. E., and N. Hugouvieux-Cotte-Pattat.
1997.
Identification of a bacterial pectin acetyl esterase in Erwinia chrysanthemi.
Mol. Microbiol.
24:1285-1301[Medline].
|
| 20.
|
Shevchik, V. E.,
J. Robert-Baudouy, and N. Hugouvieux-Cotte-Pattat.
1997.
Pectate lyase PelI of Erwinia chrysanthemi 3937 belongs to a new family.
J. Bacteriol.
179:7321-7330[Abstract/Free Full Text].
|
| 21.
|
Shevchik, V. E.,
H. C. M. Kester,
J. A. E. Benen,
J. Visser,
J. Robert-Baudouy, and N. Hugouvieux-Cotte-Pattat.
1999.
Characterization of the exopolygalacturonate lyase, PelX, of Erwinia chrysanthemi 3937.
J. Bacteriol.
181:1652-1663[Abstract/Free Full Text].
|
| 22.
| Shevchik, V. E., G. Condemine, J. Robert-Baudouy,
and N. Hugouvioux-Cotte-Pattat. The exopolygalacturonate lyase
PelW and the oligogalacturonate lyase Ogl, two cytoplasmic enzymes of
the pectin catabolism in Erwinia chrysanthemi 3937. J. Bacteriol, in press.
|
| 23.
|
Tardy, F.,
W. Nasser,
J. Robert-Baudouy, and N. Hugouvieux-Cotte-Pattat.
1997.
Comparative analysis of the five major Erwinia chrysanthemi pectate lyases: enzyme characteristics and potential inhibitors.
J. Bacteriol.
179:2503-2511[Abstract/Free Full Text].
|
| 24.
|
Yoder, M. D.,
N. T. Keen, and F. Jurnak.
1993.
New domain motif: the structure of pectate lyase C, a secreted plant virulence factor.
Science
260:1503-1507[Abstract/Free Full Text].
|
| 25.
|
Yoder, M. D., and F. Jurnak.
1995.
The refined three-dimensional structure of pectate lyase C from Erwinia chrysanthemi at 2.2 Angstrom resolution.
Plant Physiol.
107:349-364[Abstract].
|
Journal of Bacteriology, June 1999, p. 3705-3709, Vol. 181, No. 12
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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Abstract
Introduction
