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show all sequences of 4.2.2.8

Biochemical analysis and kinetic modeling of the thermal inactivation of MBP-fused heparinase I: implications for a comprehensive thermostabilization strategy

Chen, S.; Ye, F.; Chen, Y.; Chen, Y.; Zhao, H.; Yatsunami, R.; Nakamura, S.; Arisaka, F.; Xing, X.H.; Biotechnol. Bioeng. 108, 1841-1851 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression of maltose-binding protein fusion HepI, MBP-HepI, in Escherichia coli
Pedobacter heparinus
Engineering
Amino acid exchange
Commentary
Organism
C297S
site-directed mutagenesis, the mutant suppresses the dimerization and shows 70% reduced activity compared to the wild-type enzyme
Pedobacter heparinus
General Stability
General Stability
Organism
after freeze-thawing, 97% of maximal activity remains
Pedobacter heparinus
Inhibitors
Inhibitors
Commentary
Organism
Structure
alpha-lactose
-
Pedobacter heparinus
Ca2+
-
Pedobacter heparinus
Dextran
-
Pedobacter heparinus
DTT
suppresses the dimerization
Pedobacter heparinus
glycerol
-
Pedobacter heparinus
PEG 20000
-
Pedobacter heparinus
PEG 8000
-
Pedobacter heparinus
sucrose
-
Pedobacter heparinus
trehalose
-
Pedobacter heparinus
Triton X-100
-
Pedobacter heparinus
Tween 80
-
Pedobacter heparinus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetic modeling based on inactivation data, overview
Pedobacter heparinus
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Ca2+
activates
Pedobacter heparinus
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pedobacter heparinus
-
formerly Flavobacterium heparinum
-
Purification (Commentary)
Commentary
Organism
recombinant maltose-binding protein fusion HepI, MBP-HepI, from Escherichia coli by affinity and anion exchange chromatography, followed by gel filtration to apparent homogeneity
Pedobacter heparinus
Renatured (Commentary)
Commentary
Organism
temperature-induced reactivation of MBP-HepI, when the temperature is lowered from 35°C to 4°C, the rate constant of unfolding decreases by 6000times while that of refolding decreases by only 600times, MBP-HepI undergoes reactivation during the cooling treatment at 4°C after incubation at 35°C
Pedobacter heparinus
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
149
-
purified recombinant HepI, pH 7.5, 30°C
Pedobacter heparinus
Storage Stability
Storage Stability
Organism
4°C, purified recombinant MBP-HEPI, 1 week, 95% remaining activity
Pedobacter heparinus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
cleavage/degradation of heparin forming unsaturated uronic acid
714553
Pedobacter heparinus
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
More
three-dimensional model structure, modelling based on the known crystal structure of Bacteroides thetaiotaomicron HepI, overview
Pedobacter heparinus
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
-
Pedobacter heparinus
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
additional information
-
the inactivation shows strong concentration dependence. Acceleration of the inactivation with the increase of enzyme concentration implies multi-molecular interactions in the thermal inactivation. Strong reversibility of the unfolding of MBP-HepI, overview
Pedobacter heparinus
30
-
purified recombinant enzyme, half-life is 160 min
Pedobacter heparinus
70
-
purified recombinant enzyme, loss of 97% activity within 1 min
Pedobacter heparinus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Pedobacter heparinus
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of maltose-binding protein fusion HepI, MBP-HepI, in Escherichia coli
Pedobacter heparinus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C297S
site-directed mutagenesis, the mutant suppresses the dimerization and shows 70% reduced activity compared to the wild-type enzyme
Pedobacter heparinus
General Stability (protein specific)
General Stability
Organism
after freeze-thawing, 97% of maximal activity remains
Pedobacter heparinus
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
alpha-lactose
-
Pedobacter heparinus
Ca2+
-
Pedobacter heparinus
Dextran
-
Pedobacter heparinus
DTT
suppresses the dimerization
Pedobacter heparinus
glycerol
-
Pedobacter heparinus
PEG 20000
-
Pedobacter heparinus
PEG 8000
-
Pedobacter heparinus
sucrose
-
Pedobacter heparinus
trehalose
-
Pedobacter heparinus
Triton X-100
-
Pedobacter heparinus
Tween 80
-
Pedobacter heparinus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetic modeling based on inactivation data, overview
Pedobacter heparinus
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Ca2+
activates
Pedobacter heparinus
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant maltose-binding protein fusion HepI, MBP-HepI, from Escherichia coli by affinity and anion exchange chromatography, followed by gel filtration to apparent homogeneity
Pedobacter heparinus
Renatured (Commentary) (protein specific)
Commentary
Organism
temperature-induced reactivation of MBP-HepI, when the temperature is lowered from 35°C to 4°C, the rate constant of unfolding decreases by 6000times while that of refolding decreases by only 600times, MBP-HepI undergoes reactivation during the cooling treatment at 4°C after incubation at 35°C
Pedobacter heparinus
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
149
-
purified recombinant HepI, pH 7.5, 30°C
Pedobacter heparinus
Storage Stability (protein specific)
Storage Stability
Organism
4°C, purified recombinant MBP-HEPI, 1 week, 95% remaining activity
Pedobacter heparinus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
cleavage/degradation of heparin forming unsaturated uronic acid
714553
Pedobacter heparinus
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
More
three-dimensional model structure, modelling based on the known crystal structure of Bacteroides thetaiotaomicron HepI, overview
Pedobacter heparinus
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
-
Pedobacter heparinus
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
additional information
-
the inactivation shows strong concentration dependence. Acceleration of the inactivation with the increase of enzyme concentration implies multi-molecular interactions in the thermal inactivation. Strong reversibility of the unfolding of MBP-HepI, overview
Pedobacter heparinus
30
-
purified recombinant enzyme, half-life is 160 min
Pedobacter heparinus
70
-
purified recombinant enzyme, loss of 97% activity within 1 min
Pedobacter heparinus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Pedobacter heparinus
General Information
General Information
Commentary
Organism
additional information
intermolecular disulfide bond formation in HepI important for catalysis, overview
Pedobacter heparinus
General Information (protein specific)
General Information
Commentary
Organism
additional information
intermolecular disulfide bond formation in HepI important for catalysis, overview
Pedobacter heparinus
Other publictions for EC 4.2.2.8
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
748820
Gu
Photoswitchable heparinase II ...
Pedobacter heparinus
Org. Lett.
20
48-51
2018
-
1
-
-
-
-
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-
-
-
-
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1
-
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1
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-
-
-
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-
747456
Hu
Probing cleavage promiscuity ...
Pedobacter heparinus
Carbohydr. Polym.
173
276-285
2017
-
-
1
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
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16
-
-
-
-
-
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-
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-
-
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-
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1
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-
-
-
-
-
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16
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
747570
Gu
Expanding the catalytic promi ...
Pedobacter heparinus
Chemistry
23
2548-2551
2017
-
-
-
-
3
-
-
5
-
-
2
-
-
1
-
-
-
-
-
-
-
-
3
-
-
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-
5
-
-
-
-
-
-
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-
3
-
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5
-
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2
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-
-
-
-
-
-
3
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
747916
Ulaganathan
Conformational flexibility of ...
Bacteroides thetaiotaomicron, Bacteroides thetaiotaomicron DSM 2079
Glycobiology
27
176-187
2017
-
-
1
1
3
-
-
1
-
-
-
-
-
4
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
3
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
747454
Carnachan
Determining the extent of hep ...
Pedobacter heparinus
Carbohydr. Polym.
152
592-597
2016
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
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-
-
-
-
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-
-
-
-
-
-
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-
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-
-
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-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
729269
Hashimoto
Crystal structure of Pedobacte ...
Pedobacter heparinus, Pedobacter heparinus DSM 2366
Biochemistry
53
777-786
2014
-
-
-
1
11
-
-
-
-
-
-
-
-
4
-
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-
-
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6
-
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-
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-
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-
1
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
730858
Dong
Structural basis of heparan su ...
Bacteroides thetaiotaomicron, Bacteroides thetaiotaomicron DSM 2079
Protein Cell
3
950-961
2012
-
-
1
1
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
-
-
-
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1
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1
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-
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-
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-
-
-
-
-
-
-
-
-
-
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-
714553
Chen
Biochemical analysis and kinet ...
Pedobacter heparinus
Biotechnol. Bioeng.
108
1841-1851
2011
-
-
1
-
1
1
11
1
-
1
-
-
-
1
-
-
1
-
1
-
1
1
1
1
1
-
3
-
1
-
-
-
-
-
-
-
-
1
-
-
1
1
-
11
-
1
-
1
-
-
-
-
-
1
1
-
1
1
1
1
1
-
3
-
1
-
-
-
-
1
1
-
-
-
701676
Babu
Fluorescent-tagged heparan sul ...
Pedobacter heparinus
Anal. Biochem.
396
124-132
2010
-
-
1
-
-
-
-
-
-
-
-
-
-
1
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1
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-
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1
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1
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1
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1
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
701885
Hyun
Cloning, overexpression, and c ...
Bacteroides stercoris, Bacteroides stercoris HJ-15
Appl. Microbiol. Biotechnol.
86
879-890
2010
-
-
1
-
4
-
3
2
-
3
1
-
-
5
-
-
1
-
-
-
5
-
6
1
1
1
1
-
1
1
-
-
-
1
-
-
-
1
-
-
4
-
-
3
-
2
-
3
1
-
-
-
-
1
-
-
5
-
6
1
1
1
1
-
1
1
-
1
-
-
-
-
-
-
714027
Raman
Differential effects of hepari ...
Pedobacter heparinus
Biochem. Biophys. Res. Commun.
398
191-193
2010
-
-
-
-
-
-
-
-
-
-
-
-
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3
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1
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1
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-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
682098
Rops
Heparan sulfate on activated g ...
Pedobacter heparinus
Nephrol. Dial. Transplant.
22
1070-1077
2007
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
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-
-
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-
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-
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-
665613
Wei
Distinct substrate specificiti ...
Pedobacter heparinus
J. Biol. Chem.
280
15742-15748
2005
-
-
-
-
-
-
-
-
-
-
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3
-
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1
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1
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-
664047
Chai
Relative susceptibilities of t ...
Pedobacter heparinus
Biochemistry
43
8590-8599
2004
-
-
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-
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2
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1
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-
649969
Pojasek
Histidine 295 and histidine 51 ...
Pedobacter heparinus
Biochemistry
39
4012-4019
2000
-
-
1
-
13
-
1
11
-
-
-
-
-
2
-
-
1
1
-
-
-
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1
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1
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12
1
-
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1
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-
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1
-
-
13
-
-
1
1
11
-
-
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1
-
-
-
-
1
-
1
-
-
12
1
-
-
-
-
-
-
-
-
-
37411
Graham
Inhibition of platelet heparit ...
Homo sapiens
Biochem. Mol. Biol. Int.
37
239-246
1995
-
-
-
-
-
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2
-
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1
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1
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1
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37409
Yamada
Structural studies on the tri- ...
Pedobacter heparinus
Glycobiology
4
69-78
1994
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
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-
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1
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-
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-
-
-
-
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-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
37410
Suhahara
Structural studies on the olig ...
Pedobacter heparinus
Glycobiology
4
535-544
1994
-
-
-
-
-
-
-
-
-
-
-
-
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4
-
-
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-
-
2
-
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-
1
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-
1
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-
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-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
37412
Brickman
Heparitinase inhibition of mes ...
Pedobacter heparinus
Dev. Biol.
164
484-501
1994
-
1
-
-
-
-
-
-
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-
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4
-
-
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-
-
1
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-
-
-
-
-
-
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-
-
1
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Pedobacter heparinus
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1987
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