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Applied and Environmental Microbiology, March 1999, p. 1343-1347, Vol. 65, No. 3
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Cloning and Characterization of a cDNA Encoding a
Novel Extracellular Peroxidase from Trametes
versicolor
Patrick J.
Collins,
Margaret
M.
O'Brien, and
Alan D. W.
Dobson*
Microbiology Department, University College
Cork, Cork, Ireland
Received 24 August 1998/Accepted 12 December 1998
 |
ABSTRACT |
The white rot basidiomycete Trametes versicolor
secretes a large number of peroxidases which are believed to be
involved in the degradation of polymeric lignin. These peroxidases have
been classified previously as lignin peroxidases or manganese
peroxidases (MnP). We have isolated a novel extracellular
peroxidase-encoding cDNA sequence from T. versicolor CU1,
the transcript levels of which are repressed by low concentrations of
Mn2+ and induced by nitrogen and carbon but not induced in
response to a range of stresses which have been reported to induce MnP expression.
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TEXT |
The lignin-degrading ability of the
white rot basidiomycete Trametes versicolor has been well
studied (2, 10). Along with laccases, two groups of
heme-containing proteins, lignin peroxidases (LiP) and manganese
peroxidases (MnP), have been reported to be involved in the
ligninolytic process. LiP attacks both phenolic and nonphenolic
aromatic residues, with the latter giving rise to cation radicals that
fragment spontaneously (24). MnP catalyzes the oxidation of
Mn(II) to Mn(III), which in turn can oxidize phenolic lignin subunits
(14). LiP and MnP are both produced by white rot fungi as a
number of isozymes which are encoded by families of structurally
related genes. In one study, 16 forms of LiP and 5 forms of MnP were
detected in a T. versicolor culture fluid
(16). A number of genomic clones encoding LiP
isozymes (21) have been characterized, and recently
Johansson and Nyman (17) have described a
genomic sequence encoding a T. versicolor MnP. In
addition, a third type of peroxidase-encoding gene, PGV, has
been isolated from T. versicolor PRL 572 (20). The expression of various isozymes of both the LiP and
MnP subfamilies is differentially regulated at the level of gene
transcription by physiological growth conditions such as carbon or
nitrogen concentration (9, 13). Brown et al. (5)
have demonstrated that production of mRNA encoding MnP in
Phanerochaete chrysosporium is induced by the enzyme's
primary substrate, Mn2+. Furthermore, Mn2+
induction of different MnP isozymes is coordinately regulated in
P. chrysosporium (9, 13, 32). Further studies
have indicated that MnP production in P. chrysosporium is
transcriptionally regulated by a number of other factors, including
heat shock (6), H2O2, chemical
stress, and molecular oxygen (26).
We report here the cloning and characterization of a novel
extracellular peroxidase-encoding cDNA sequence (npr) from
T. versicolor. The deduced amino acid sequence of the NPR
protein contains the 10 invariant residues of all members of the plant
peroxidase superfamily (41) (see Fig. 1). A comparison of
the sequence with various MnP and LiP sequences reveals a high degree
of similarity near the active site. In addition, the proposed
Mn2+-binding residues of MnP are present in the deduced
NPR. In contrast, the LiP residues suggested to interact with aromatic
substrates are not present. This putative protein may therefore
represent a new class of ligninolytic peroxidases. Using a reverse
transcription (RT)-PCR approach, we have determined that, in contrast
to MnP-encoding genes, npr transcript levels are repressed
by low levels of Mn2+. In addition, we have investigated
the effects on npr transcript levels of a number of other
factors which are known to regulate MnP and LiP expression.
Cloning and characterization of the npr cDNA
sequence.
A Lambda ZAP II T. versicolor CU1 cDNA
library, constructed by Stratagene (Cambridge, United Kingdom) and
containing 3.3 × 109 amplified recombinants per ml,
was plated at a density of 5 × 104 PFU per 150-mm
petri dish and hybridized overnight at 55°C with 32P-labelled MNP-1 from P. chrysosporium (34), as previously described (37). After hybridization, the membranes were washed once
with 2× SSC (1× SSC is 0.15 M NaCl plus 0.015 M sodium
citrate)-0.1% sodium dodecyl sulfate for 20 min at 55°C and twice
with 0.2× SSC-0.1% sodium dodecyl sulfate for 20 min at 55°C. Six
putative positive clones were isolated, and one of these,
npr, was chosen for sequencing and characterization.
Sequencing was via the dideoxy chain termination method
(38), with a Dye Terminator Cycle Sequencing Ready Reaction
Kit and AmpliTaq DNA polymerase FS (Applied Biosystems, Warrington, United Kingdom), on a GeneAmp PCR System 2400 (Perkin-Elmer, Norwalk, Conn.) and an automated DNA sequencer (model
373 Stretch; Applied Biosystems). The sequencing strategy consisted of
the initial use of M13 universal primers, followed by designing of primers by using the newly acquired sequences. Both strands were sequenced independently. Sequence data was assembled and processed with
the DNASTAR (DNASTAR, Inc., Madison, Wis.) software package. The BLAST
algorithm (1) was used to search DNA and protein databases
for similarity. The CLUSTAL program was used for amino acid sequence alignment.
The npr nucleotide sequence is 1,362 bp long and contains an
open reading frame of 1,089 bp which encodes a protein of 362 amino
acids. The coding region G+C content is 66%, similar to that
previously reported for T. versicolor LiP- and MnP-encoding exon regions (17). The predicted mature NPR translation
product is shown in Fig. 1. Eight
cysteine residues are present within the deduced NPR sequence, allowing
four possible disulfide linkages to be formed. This would be consistent
with evidence from X-ray crystallographic studies with LiP
(33) from P. chrysosporium, which indicates an
arrangement of four disulfide linkages of 1-3, 2-7, 4-5, and 6-8. Such
linkages have also been observed in peroxidases from T. versicolor (27). However, this differs from MnP, which has a fifth disulfide bond believed to be required for the correct formation of the Mn2+ binding site (39).
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FIG. 1.
Comparison of the NPR amino acid sequence with
various fungal manganese and lignin peroxidases. Areas of dark
background indicate common amino acids. The positions of the amino
acids involved in Mn2+ binding (19, 25, 39) are
indicated with squares. The 10 invariant residues of all members of the
plant peroxidase superfamily (41) are indicated with
inverted triangles. The positions of the LiP residues suggested to
interact with aromatic substrates are indicated by circles
(33). The amino acid sequences were either experimentally
determined or deduced from nucleotide sequences of T. versicolor (Tv NPR [this study] and Tv PGV
[20]), P. chrysosporium manganese
peroxidase (Pc MNP1) (34), Ceriporiopsis
subvermispora manganese peroxidase (Cs MNPI) (28),
P. ostreatus manganese peroxidase (Po MNPII) (4),
T. versicolor lignin peroxidase (Tv LPGIII) (17),
P. chrysosporium lignin peroxidase (Pc LPO811)
(35), Phlebia radiata lignin peroxidase (Pr LPG3)
(36), and B. adusta lignin peroxidase (Ba LPO)
(3).
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A phylogenetic tree was constructed, based on the alignment of the
npr-encoded amino acid sequence and amino acid sequences
deduced for LiP and MnP from
T. versicolor and
P. chrysosporium (data not shown). This indicates that NPR is
significantly distant
at the amino acid level from both the MnP and LiP
subfamilies
within the plant peroxidase superfamily proposed by
Welinder (
40).
It shows an identity of approximately 45 to
50% in amino acid
sequence with both LiP and MnP sequences from
T. versicolor and
no more than 53.6% identity with any of
the other sequences examined.
In the MnP subfamily, NPR appears to be
most closely related to
PGV (
20), a
T. versicolor peroxidase which displays a number
of novel features.
Within these groups, NPR exhibits the greatest
divergence from
P. chrysosporium MnP
sequences.
Analysis of npr transcript levels.
In order to
determine the point of maximal npr expression, a time course
experiment in which npr transcript levels were measured after 4, 6, 8, 10, 12, 14, and 18 days of growth was conducted. An
RT-PCR approach which included an internal control system standardizing both the RT and PCR processes was employed. Total RNA was prepared from
mycelium samples, as previously described (7), with residual contaminating DNA being removed by digestion with DNase I. Total RNA
was quantitated by both spectrophotometric and visual methods (37). RT reaction mixtures contained 1 µg of total RNA,
random hexamer primers, and other reaction components previously
described (8). In order to standardize each set of RT-PCR
mixtures, a known amount (10 pg) of a control template was added to
each RT reaction mixture. This control consisted of RNA transcripts
generated from a genomic lignin peroxidase gene
(lip) fragment with the Riboprobe in vitro transcription
system (Promega). The DNA template used in this in vitro transcription
reaction had been amplified from T. versicolor CU1
genomic DNA with the lip forward
(5'-CGACGCIATCGCCATCTC-3' [I represents inosine]) and
reverse (5'-GAACGGCTCGGG[G/C]ACGAG-3') primers previously
described (8). This fragment contains two introns, yielding
an RT-PCR product of 418 bp, and thus could be distinguished from a
fragment representing lip mRNA which would, if present, be
304 bp in size. The RT reaction mixtures were incubated at 37°C for
1 h, and reactions were terminated by heating to 65°C for 10 min. PCR amplifications were carried out with forward
(5'-TCATGGCCCACACCGACG-3') and reverse
(5'-GCGGTCGGCGATCATCTT-3') primers designed to specifically amplify npr cDNA sequences and not those encoding either LiP
or MnP. With these primers, the npr amplification
product is 560 bp. For PCR amplification, a 2-µl volume from each RT
reaction was mixed with 75 ng of each primer, 5 µl of 10×
NH4Cl Taq buffer (Bioline, London, United
Kingdom), 1.5 mM MgCl2, 100 µM (each) deoxynucleoside
triphosphates, and 1.25 U of Taq polymerase. In addition,
each reaction mixture contained 75 ng of both lip PCR primers (8) in order to amplify the control RT-PCR fragment in each case. For each experiment, the constant band intensities seen
for the amplified product of this lip fragment (see Fig. 2
to 5, lower bands) indicated the uniformity of RT-PCR efficiency. Reaction volumes were adjusted to 50 µl with water. Amplification was
performed in a PTC-100 programmable thermal controller (MJ Research,
Inc., Watertown, Mass.) with 30 cycles of denaturation (1 min at
94°C), annealing (1 min at 56°C), and extension (1 min at 72°C).
Ten-microliter aliquots of each reaction mixture were electrophoresed
on 2% agarose gels, and the product band intensities within each
experiment were visually compared after ethidium bromide staining. Both
the npr fragment and the lip control fragment
were sequenced to confirm their identities.
The time course experiment indicated the presence of
npr
transcripts after 4 days of growth, reaching a maximum after 8 days
and
then beginning to decrease. However,
npr mRNA was still
detected
in 18-day-old cultures (Fig.
2).
As the highest level of
npr transcription
was observed on
day 8, for subsequent experiments, fungal mycelia
from quadruplicate
cultures were harvested, pooled, and analyzed
for each datum point
following 8 days of incubation.
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FIG. 2.
Time course of npr transcription in cultures
of T. versicolor CU1. The higher-molecular-weight band (560 bp) represents the level of npr transcript present, and the
lower band (418 bp) represents the lip fragment amplified as
an internal control.
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The effects of Mn
2+ on
npr transcript levels
were investigated by adding MnSO
4 at final concentrations
of 0.5, 2, 20, and 200
µM to the growth medium. In order to remove
trace levels of Mn
2+ from the media used, all glassware was
soaked overnight in 2
M nitric acid and then extensively rinsed with
distilled deionized
water. Mn
2+ levels in media were
measured by flame atomic absorption spectroscopy
(Pye Unicam,
Cambridge, United Kingdom) at a resonance line of
279.5 nm and in all
cases were found to be lower than the detection
limit of the instrument
(0.018 µM). Supplementation of fungal
cultures with low levels of
Mn
2+ had a repressive effect on
npr transcript
levels (Fig.
3). A
high level of
amplification product, representing
npr mRNA, was
observed
when the medium contained no detectable Mn
2+. However when
Mn
2+ at a concentration of 0.5 µM was added, a much
reduced level
of
npr transcript was detected. A further
decrease in the level
of
npr transcription occurred when the
Mn
2+ concentration was increased to 2 µM. Interestingly
however, a
low level of
npr transcription was still detected
at Mn
2+ concentrations as high as 200 µM and even up to
500 µM (data
not shown).
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FIG. 3.
Effects of growth in the presence of various
concentrations of Mn2+ on npr transcription
levels. The higher-molecular-weight band (560 bp) represents the level
of npr transcript present, and the lower band (418 bp)
represents the lip fragment amplified as an internal
control.
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The possibility that expression of
npr may be induced as a
response to environmental stresses was investigated by subjecting
8-day-old cultures to a range of stresses before analysis. This
was
examined by adding either 2,5-xylidine (at a final concentration
of 5, 20, 50, 200, or 500 µM) or ethanol (at a final concentration
of 1, 2, 5, 10, or 15%) to 8-day-old cultures. For these experiments,
cultures
were harvested for analysis at 90 min after addition
of the compounds.
The effect of H
2O
2 addition (at a final
concentration
of 0.2, 0.5, 1, 2, or 5 mM) was determined by adding it
to 8-day-old
cultures for 80 min. For determination of a possible
regulatory
effect of heat shock, cultures were transferred from 26°C
to water
baths at either 37 or 45°C for a period of 120 min prior to
harvesting
of the mycelium. No effect on the level of
npr
transcription was
observed as a response to either heat shock,
oxidative stress
in the form of H
2O
2, or
chemical stress in the form of ethanol
(data not shown). The presence
of 2,5-xylidine, however, which
previously has been demonstrated to
induce laccase gene transcription
in
T. versicolor
(
7), was found to have a repressive effect
on
npr
expression when present at concentrations as low as 5 µM
(data not
shown). This phenomenon may be due simply to the toxic
effect of the
compound on cellular
metabolism.
Experiments were also conducted to determine the effect of nitrogen on
npr transcript levels by culturing the fungus on basal
medium (
8) containing ammonium sulfate as the nitrogen
source
and no MnSO
4. Expression of
npr was found
to be induced by nitrogen
at the level of gene transcription (Fig.
4). As the concentration
of nitrogen was
increased from 2 mM (limiting nitrogen) to 5 mM
(high nitrogen), a
corresponding increase in
npr transcript levels
was
observed. A similar effect was observed when the glucose concentration
was observed when the glucose concentration was increased from
0.5 to
2 g per liter (Fig.
5). Further
increases in the level
of glucose in the medium had a less marked
inducing effect.
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FIG. 4.
Effects of growth in the presence of various
concentrations of nitrogen on npr transcription levels. The
higher-molecular-weight band (560 bp) represents the level of
npr transcript present, and the lower band (418 bp)
represents the lip fragment amplified as an internal
control.
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FIG. 5.
Effects of growth in the presence of various
concentrations of carbon in the form of glucose on npr
transcription levels. The higher-molecular-weight band (560 bp)
represents the level of npr transcript present, and the
lower band (418 bp) represents the lip fragment amplified as
an internal control.
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The deduced amino acid sequence of the NPR protein isolated here
exhibits features characteristic of
ligninolytic peroxidases
from other
strains of
T. versicolor and other white rot fungal
species.
NPR also displays a high degree of amino acid sequence
similarity, at
and near its active site, with MnP, LiP, and a
range of other
peroxidases, i.e., the regions surrounding the
so-called distal His
(residue 47), believed to be involved in
charge stabilization during
reaction of the heme prosthetic group
with
H
2O
2, and proximal His (residue 175) residue,
the proposed
axial ligand of heme. In addition, an Arg (residue 43)
also essential
for activity (
11) and a Leu (residue 44) near
the distal His
are well conserved in NPR and the various other MnP and
LiP amino
acid sequences. Furthermore, the deduced NPR amino acid
sequence
contains the 10 invariant residues of all members of the plant
peroxidase superfamily (
41) (Fig.
1).
Alignment of the
npr DNA sequence (data not shown) or its
deduced amino acid sequence with corresponding sequences for
ligninolytic
peroxidases indicated that NPR cannot be classified as
either
an LiP- or an MnP-type peroxidase. Instead, the product of
npr expression may represent a more distinct class of
peroxidase within
the
ligninolytic peroxidase family. The deduced
NPR protein does,
however, display some unique features which suggest a
closer structural
and functional relationship with MnP than with
LiP. Firstly, the
total Ser content of deduced NPR (26 residues)
is consistent with
the reported value for MnP (approximately 24 residues) rather
than that for LiP (16 residues) (
18).
Johansson et al. (
18)
have previously reported this
difference in the overall serine
content in eight different LiP and MnP
isozymes from
T. versicolor.
Secondly, a number of key
residues in MnP, proposed to interact
with its Mn
2+
substrate (
19,
39), are present in the deduced NPR sequence
(Glu
36, Glu
40, and Asp
181). In
contrast, the LiP residues suggested to interact with aromatic
substrates of the enzyme (
33) are not present in the NPR
sequence
(Fig.
1). This suggests that NPR may be evolutionarily, or
even
functionally, more closely related to MnP. Although possibly
related
to MnP at the functional level, NPR expression contrasts
markedly
with regard to its regulation by Mn
2+.
Mn
2+ has been demonstrated to directly induce
mnp transcription in
P. chrysosporium
(
5,
12,
13), and this is thought to be
mediated via a metal
response element in the
mnp promoter region
(
15).
In the case of
npr, however, Mn
2+ appears to
have a similar but opposite effect whereby the presence
of low
concentrations of Mn
2+ have a repressive effect on
transcription.
A number of studies have reported that some level of
mnp
transcription occurs in
P. chrysosporium in the absence
of Mn
2+ (
5) and in the presence or absence of
some other
mnp-inducing
factor (
6,
26).
Furthermore, Moreira and coworkers (
31)
reported that MnP
was the predominant oxidative enzyme produced
by
Bjerkandera
sp. strain BOS55, even under Mn
2+-deficient conditions.
This group has recently reported the characterization
of an MnP-LiP
hybrid enzyme produced by this strain in the absence
of manganese
(
30).
Further contrasts also exist between the regulation of
npr
and
mnp. Transcription of
mnp can be induced as a
heat shock-like
response to chemical stress by ethanol, oxidative
stress by H
2O
2 (
26), or heat shock
itself (
6) in
P. chrysosporium. However,
in
our study such responses were not detected, indicating further
the
heterogeneity between
mnp and
npr with respect to
their regulation.
Increased nitrogen and carbon levels result in
increased
npr transcripts
in
T. versicolor CU1.
In
P. chrysosporium, both MnP and LiP are
produced
during secondary metabolism only under conditions of
carbon or nitrogen
limitation (
15), and various isozymes have
been shown to be
differentially regulated by these nutrients (
13,
32). In
contrast, a number of strains of
Bjerkandera adusta in which
nitrogen appears to have a stimulatory effect on both
LiP and MnP
production have been isolated (
23,
29). MnP induction
by
high nitrogen levels has also been reported in
Pleurotus
ostreatus (
23).
In conclusion, the translation product of
npr cannot
be clearly classified as an LiP- or MnP-type peroxidase
on the basis
of DNA or deduced amino acid sequence. An apparent
biological
contradiction exists in that, although it contains the
residues
proposed to function as Mn
2+-binding ligands in
MnP, transcription of
npr is repressed by
the presence
of Mn
2+. However, it is possible that these residues are
involved in
the binding of alternative substrates, such as
aromatic amines.
Furthermore, the repression of
npr
transcription by Mn
2+ makes it difficult to speculate on
what the physiological role
of its putative translation product might
be. One possibility
is that it is expressed in the later stages of
lignin degradation,
when much of the Mn
2+ present in wood
has been oxidized by MnP to MnO
2 and functions
in the
further oxidation of
lignin subunits. Future work, therefore,
will
focus on the purification of this putative NPR protein, in
either the
native form or a recombinant form, in order to characterize
it with
respect to its substrate range and catalytic mechanism
compared to MnP
and
LiP.
Nucleotide sequence accession number.
The nucleotide sequence
of the npr gene has been assigned GenBank accession no.
AF008585.
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ACKNOWLEDGMENTS |
This work was supported through the European Union FAIR Programme
Oxidative Enzymes for the Pulp and Paper Industry (OXEPI) CT95-0805.
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FOOTNOTES |
*
Corresponding author. Mailing address: Microbiology
Department, University College Cork, Cork, Ireland. Phone:
353-21-902743. Fax: 353-21-903101. E-mail:
a.dobson{at}ucc.ie.
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REFERENCES |
| 1.
|
Altschul, S. F.,
W. Gish,
W. Miller,
E. W. Myers, and D. J. Lipman.
1990.
Basic local alignment search tool.
J. Mol. Biol.
215:403-410[Medline].
|
| 2.
|
Archibald, F. S.
1993.
Lignin peroxidase activity is not important in biological bleaching and delignification of unbleached kraft pulp by Trametes versicolor.
Appl. Environ. Microbiol.
58:3101-3109[Abstract/Free Full Text].
|
| 3.
|
Asada, Y.,
Y. Kimura,
T. Oka, and M. Kuwahara.
1992.
Characterization of a cDNA and gene encoding a lignin peroxidase from the lignin-degrading basidiomycete Bjerkandera adusta, p. 421-426.
In
M. Kuwahara, and M. Shimada (ed.), Biotechnology in the pulp and paper industry. Uni Publishers, Tokyo, Japan.
|
| 4.
|
Asada, Y.,
A. Watanabe,
T. Irie,
T. Nakayama, and M. Kuwahara.
1995.
Structures of genomic and complementary DNAs coding for Pleurotus ostreatus manganese (II) peroxidase.
Biochim. Biophys. Acta.
1251:205-209[Medline].
|
| 5.
|
Brown, J. A.,
M. Alic, and M. H. Gold.
1991.
Manganese peroxidase gene transcription in Phanerochaete chrysosporium: activation by manganese.
J. Bacteriol.
173:4101-4106[Abstract/Free Full Text].
|
| 6.
|
Brown, J. A.,
D. Li,
M. Alic, and M. H. Gold.
1993.
Heat shock induction of manganese peroxidase gene transcription in Phanerochaete chrysosporium.
Appl. Environ. Microbiol.
59:4295-4299[Abstract/Free Full Text].
|
| 7.
|
Collins, P. J., and A. D. W. Dobson.
1997.
Regulation of laccase gene transcription in Trametes versicolor.
Appl. Environ. Microbiol.
63:3444-3450[Abstract].
|
| 8.
|
Collins, P. J.,
J. A. Field,
P. Teunissen, and A. D. W. Dobson.
1997.
Stabilization of lignin peroxidases in white-rot fungi by tryptophan.
Appl. Environ. Microbiol.
63:2543-2548[Abstract].
|
| 9.
|
Cullen, D.
1997.
Recent advances on the molecular genetics of ligninolytic fungi.
J. Biotechnol.
53:273-289[Medline].
|
| 10.
|
Eriksson, K.-E., L.,
R. A. Blanchette, and P. Ander.
1990.
Biochemistry of lignin degradation, p. 225-333.
In
K.-E. L. Eriksson, R. A. Blanchette, and P. Ander (ed.), Microbial and enzymatic degradation of wood and wood components. Springer-Verlag, Berlin, Germany.
|
| 11.
|
Finzel, B. C.,
T. L. Poulos, and J. Kraut.
1987.
Crystal structure of yeast cytochrome c peroxidase refined at 1.7 A resolution.
J. Biol. Chem.
259:13027-13036[Abstract/Free Full Text].
|
| 12.
|
Gettemy, J. M.,
D. Li,
M. Alic, and M. H. Gold.
1997.
Truncated gene reporter system for studying the regulation of manganese peroxidase expression.
Curr. Genet.
3:519-524.
|
| 13.
|
Gettemy, J. M.,
B. Ma,
M. Alic, and M. H. Gold.
1998.
Reverse transcription-PCR analysis of the regulation of the manganese peroxidase gene family.
Appl. Environ. Microbiol.
64:569-574[Abstract/Free Full Text].
|
| 14.
|
Glenn, J. K.,
L. Akileswaren, and M. H. Gold.
1986.
Mn(II) oxidation is the principal function of the extracellular Mn-peroxidase from Phanerochaete chrysosporium.
Arch. Biochem. Biophys.
251:688-696[Medline].
|
| 15.
|
Gold, M. H., and M. Alic.
1993.
Molecular biology of the lignin-degrading basidiomycete Phanerochaete chrysosporium.
Microbiol. Rev.
57:605-622[Abstract/Free Full Text].
|
| 16.
|
Johansson, T., and P. O. Nyman.
1993.
Isozymes of lignin peroxidase and manganese (II) peroxidase from the white-rot basidiomycete Trametes versicolor. I. Isolation of enzyme forms and characterization of physical and catalytic properties.
Arch. Biochem. Biophys.
300:49-56[Medline].
|
| 17.
|
Johansson, T., and P. O. Nyman.
1996.
A cluster of genes encoding major isozymes of lignin peroxidase and manganese peroxidase from the white-rot fungus Trametes versicolor.
Gene
170:31-38[Medline].
|
| 18.
|
Johansson, T.,
K. G. Welinder, and P. O. Nyman.
1993.
Isozymes of lignin peroxidase and manganese (II) peroxidase from the white-rot basidiomycete Trametes versicolor. II. Partial sequences, peptide maps and amino acid and carbohydrate compositions.
Arch. Biochem. Biophys.
300:57-62[Medline].
|
| 19.
|
Johnson, F.,
G. H. Loew, and P. Du.
1993.
Prediction of Mn2+ binding site of manganese peroxidase from homology modelling, p. 31-34.
In
K. G. Welinder, S. K. Rasmussen, C. Penel, and H. Greppin (ed.), Plant peroxidases: biochemistry and physiology. University of Copenhagen and Unversity of Geneva, Geneva, Switzerland.
|
| 20.
|
Jönsson, L.,
H. G. Becker, and P. O. Nyman.
1994.
A novel type of peroxidase gene from the white-rot fungus Trametes versicolor.
Biochim. Biophys. Acta
1207:255-259[Medline].
|
| 21.
|
Jönsson, L., and P. O. Nyman.
1992.
Characterization of lignin peroxidase gene from the white-rot fungus Trametes versicolor.
Biochemie
74:177-183[Medline].
|
| 22.
|
Jönsson, L., and P. O. Nyman.
1994.
Tandem lignin peroxidase genes of the white-rot fungus Trametes versicolor.
Biochim. Biophys. Acta
1218:408-412[Medline].
|
| 23.
|
Kaal, E. E. J.,
J. A. Field, and T. W. Joyce.
1995.
Increasing ligninolytic enzyme activities in several white-rot basidiomycetes by nitrogen-sufficient media.
Bioresour. Technol.
53:133-139.
|
| 24.
|
Kersten, P. J.,
M. Tien,
B. Kalyanamaran, and T. K. Kirk.
1985.
the ligninase of Phanerochaete chrysosporium generates cation radicals from methoxybenzenes.
J. Biol. Chem.
260:2609-2612[Abstract/Free Full Text].
|
| 25.
|
Kishi, K.,
M. Kusters-van Someren,
M. B. Mayfield,
J. Sun,
T. M. Loehr, and M. H. Gold.
1996.
Characterization of manganese (II) binding site mutants of manganese peroxidase.
Biochemistry
35:8986-8994[Medline].
|
| 26.
|
Li, D.,
M. Alic,
J. A. Brown, and M. H. Gold.
1995.
Regulation of manganese peroxidase gene transcription by hydrogen peroxide, chemical stress, and molecular oxygen.
Appl. Environ. Microbiol.
61:341-345[Abstract].
|
| 27.
|
Limongi, P.,
M. Kjalke,
J. Vend,
J. W. Tams,
T. Johansson, and K. G. Welinder.
1995.
Disulphide bonds and glycosylation in fungal peroxidases.
Eur. J. Biochem.
227:270-276[Medline].
|
| 28.
|
Lobos, S.,
L. Lorondo,
L. Salas,
E. Karahanian, and R. Vicuna.
1998.
Cloning and molecular analysis of a cDNA encoding a manganese peroxidase isozyme from the lignin degrading basidiomycete Ceriporiopsis subvermispora.
Gene
206:185-193[Medline].
|
| 29.
|
Mester, T.,
M. Peña, and J. A. Field.
1996.
Nutrient regulation of extracellular peroxidases in the white-rot fungus Bjerkandera sp. strain BOS55.
Appl. Microbiol. Biotechnol.
44:778-784.
|
| 30.
|
Mester, T., and J. A. Field.
1988.
Characterization of a novel manganese peroxidase-lignin peroxidase hybrid isozyme produced by Bjerkandera sp. strain BOS55 in the absence of manganese.
J. Biol. Chem.
273:15412-15417[Abstract/Free Full Text].
|
| 31.
|
Moreira, M. T.,
G. Feijoo,
R. Sierra-Alvarez,
J. Lema, and J. A. Field.
1997.
Manganese is not required for biobleaching of oxygen-delignified kraft pulp by the white rot fungus Bjerkandera sp. strain BOS55.
Appl. Environ. Microbiol.
63:1749-1755[Abstract].
|
| 32.
|
Pease, E. A., and M. Tien.
1992.
Heterogeneity and regulation of manganese peroxidases from Phanerochaete chrysosporium.
J. Bacteriol.
174:3532-3540[Abstract/Free Full Text].
|
| 33.
|
Poulos, T. L.,
S. Edwards,
H. Wariishi, and M. H. Gold.
1993.
Crystallographic refinement of lignin peroxidase at 2A.
J. Biol. Chem.
268:4429-4440[Abstract/Free Full Text].
|
| 34.
|
Pribnow, D.,
M. B. Mayfield,
V. J. Nipper,
J. A. Brown, and M. H. Gold.
1989.
Characterization of a cDNA encoding manganese peroxidase from the lignin-degrading basidiomycete Phanerochaete chrysosporium.
J. Biol. Chem.
264:5036-5040[Abstract/Free Full Text].
|
| 35.
|
Reiser, J.,
I. S. Walther,
C. Fraefel, and A. Fiecther.
1993.
Methods to investigate the expression of lignin peroxidase genes by the white rot fungus Phanerochaete chrysosporium.
Appl. Environ. Microbiol.
59:2897-2903[Abstract/Free Full Text].
|
| 36.
|
Saloheimo, M.,
V. Barajas,
M.-L. Niko-Paavola, and J. K. C. Knowles.
1989.
A lignin-peroxidase encoding cDNA from the white-rot fungus Phlebia radiata: characterization and expression in Trichoderma reesei.
Gene
85:343-351[Medline].
|
| 37.
|
Sambrook, J.,
E. F. Fritsch, and T. Maniatis.
1989.
Molecular cloning: a laboratory manual, 2nd ed.
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
|
| 38.
|
Sanger, F.,
S. Nicklen, and A. R. Coulson.
1977.
DNA sequencing with chain-terminating inhibitors.
Proc. Natl. Acad. Sci. USA
74:5463-5467[Abstract/Free Full Text].
|
| 39.
|
Sundaramoorthy, M.,
K. Kishi,
M. H. Gold, and T. L. Poulos.
1994.
The crystal structure of manganese peroxidase from Phanerochaete chrysosporium at 2.06-A resolution.
J. Biol. Chem.
269:32759-32767[Abstract/Free Full Text].
|
| 40.
|
Welinder, K. G.
1992.
Superfamily of plant, fungal and bacterial peroxidases.
Curr. Opin. Struct. Biol.
2:383-393.
|
| 41.
|
Welinder, K. G., and M. Gajhede.
1993.
Structure and evolution of peroxidases, p. 35-42.
In
K. G. Welinder, S. K. Rasmussen, C. Penel, and H. Greppin (ed.), Plant peroxidases: biochemistry and physiology. University of Copenhagen and University of Geneva, Geneva, Switzerland.
|
Applied and Environmental Microbiology, March 1999, p. 1343-1347, Vol. 65, No. 3
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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