Information on EC 3.2.1.81 - beta-agarase

agar + H2O

neoagarobiose + ?

-

-

?

agar + H2O

neoagarobiose + neoagarotetraose + neoagarohexaose

8 entries

agar + H2O

neoagarohexaose + neoagarotetraose

agar + H2O

neoagarooctaose + neoagarohexaose + neoagarotetraose

-

-

?

agar + H2O

neoagarotetraose + neoagarohexaose

agaropectin + H2O

?

-

-

ir

agarose + H2O

2 neoagarooligosaccharides

rAgaC with low activity (0.05 U/ml) cleaves agarose into longer neoagarooligosaccharides. When activity is increased to 30 U/ml, neoagarooctaose is produced predominantly in addition to neoagarohexaose and neoagarotetraose

-

?

agarose + H2O

?

10 entries

agarose + H2O

agarotetraose + ?

agarose + H2O

neoagarobiose

8 entries

agarose + H2O

neoagarobiose + neoagarotetraose

agarose + H2O

neoagarobiose + neoagarotetraose + neoagarohexaose

agarose + H2O

neoagarohexaose + neoagarotetraose

agarose + H2O

neoagarohexaose + neoagarotetraose + neoagarobiose

agarose + H2O

neoagarooctaose + ?

agarose + H2O

neoagarooctaose + neoagarodecaose

agarose + H2O

neoagarosebiose

agarose + H2O

neoagarotetraose

25 entries

agarose + H2O

neoagarotetraose + ?

agarose + H2O

neoagarotetraose + neoagarobiose

10 entries

agarose + H2O

neoagarotetraose + neoagarohexaose

16 entries

agarose + H2O

neoagarotetraose + neoagarohexaose + neoagarobiose

agarose + H2O

neoagarotetraose + neoagarohexaose + neoagarooctaose

alginate + H2O

?

-

-

?

alkali treated porphyran + H2O

?

carboxymethyl cellulose + H2O

?

carrageenan + H2O

?

-

-

?

chitin + H2O

?

Gracilaria lemaneiformis + H2O

neoagarobiose + neoagarotetraose

-

-

?

liquid-phase agarose + H2O

?

-

-

?

neoagaro-oligosaccharides + H2O

?

-

from N2 to N22, reaction yield for neoagaro-oligosaccharides is 52.7% by 4 U/mg beta-agarase

697125

-

?

neoagarodecaose + H2O

?

is a very poor substrate

-

?

neoagarodecaose + H2O

neoagarobiose

neoagarododecaose + H2O

?

-

-

?

neoagarododecaose + H2O

neoagarohexaose + neoagarotetraose + neoagarooctaose

-

-

?

neoagarohexaitol + H2O

?

-

136878

-

ir

neoagarohexaose + H2O

?

neoagarohexaose + H2O

neoagarobiose

neoagarohexaose + H2O

neoagarobiose + neoagarotetraose

neoagarohexaose + H2O

neoagarotetraose + neoagarobiose

neoagarooctaose + H2O

neoagarobiose

neoagarooctaose + H2O

neoagarobiose + neoagarotetraose + neoagarohexaose

-

-

?

neoagarooctaose + H2O

neoagarohexaose + neoagarobiose

neoagarooctaose + H2O

neoagarohexaose + neoagarotetraose + neoagarobiose

neoagarooctaose + H2O

neoagarotetraose

neoagarotetradecaose + H2O

?

-

-

?

neoagarotetraose + H2O

neoagarobiose

polysaccharides with neoagarobiose units + H2O

neoagarobiose + neoagarotetraose

-

-

ir

porphyran + H2O

neoagaro-oligosaccharides

porphyran + H2O

neoagarohexaose + neoagarotetraose

porphyran + H2O

neoagarooctaose + neoagarohexaose + neoagarotetraose

-

-

?

xylan + H2O

?

additional information

?

-

21 entries

agar + H2O

neoagarobiose + ?

-

-

?

agar + H2O

neoagarobiose + neoagarotetraose + neoagarohexaose

agar + H2O

neoagarotetraose + neoagarohexaose

agarose + H2O

neoagarobiose

agarose + H2O

neoagarobiose + neoagarotetraose + neoagarohexaose

agarose + H2O

neoagarohexaose + neoagarotetraose + neoagarobiose

agarose + H2O

neoagarooctaose + ?

agarose + H2O

neoagarooctaose + neoagarodecaose

Escherichia coli

-

major products

-

?

agarose + H2O

neoagarotetraose

Ba2+

Ca2+

CaCl2

Co2+

-

135% activity at 1 mM

Cu2+

-

1 mM, increases activity of agarase AG-b 1.2fold

Fe2+

2 mM stimulates agarase activity by 20%

K+

KCl

-

no activity in the presence of 00.3 M salt. The enzyme shows a salt requirement for activity, being active from 0.3 M NaCl, with maximal activity at 3.5 M NaCl. KCl supports similar activities as NaCl up to 3.5 M, and LiCl up to 2.5 M. The monovalent salts can not be substituted by 3.5 M divalent cations, CaCl2 or MgCl2

LiCl

-

no activity in the presence of 00.3 M salt. The enzyme shows a salt requirement for activity, being active from 0.3 M NaCl, with maximal activity at 3.5 M NaCl. KCl supports similar activities as NaCl up to 3.5 M, and LiCl up to 2.5 M. The monovalent salts can not be substituted by 3.5 M divalent cations, CaCl2 or MgCl2

Mg2+

Mn2+

Na+

NaCl

additional information

1-ethyl-(3-(3-dimethyl-amino-)propyl)carbonate

-

1 mM. 10-20% inhibition

5,5'-dithiobis-(2-nitrobenzoic acid)

Agarivorans sp. LQ48

C9WIW7

53% residual activity at 10 mM

705482

Ag+

-

5 mM, complete inhibition of agarase AG-b

AgNO3

Al3+

AlCl3

Ba2+

-

85% residual activity at 1 mM

Ca2+

Cd2+

-

about 85% residual activity at 2 mM

Co2+

Cu2+

CuCl2

CuSO4

diethyl dicarbonate

-

1 mM, 10-20% inhibition

EDTA

11 entries

EGTA

-

the inhibition by 1 mM EGTA is almost completely recovered by the addition of Co2+ at a concentration of 0.2 mM

Fe2+

Fe3+

FeCl2

-

2 mM, 99% inhibition

656317

FeCl3

-

partially inhibits the activity

Hg2+

HgCl2

K+

-

94.9% residual activity at 100 mM

701877

KCl

-

50% inhibition at 0.1 M

L-galactose 6-sulfate

-

Mg2+

Mn2+

MnCl2

-

partially inhibits the activity

N-bromosuccinimide

NaCl

NH4Cl

-

50% inhibition at 0.1 M

Ni2+

90.26% residual activity at 10 mM

Pb(CH3COO)2

Pb2+

PbCl2

-

2 mM, 98% inhibition

656317

SDS

sodium dodecylsulfate

Urea

85.86% residual activity at 10 mM

Zn2+

ZnCl2

ZnSO4

additional information

9 entries

-

2-mercaptoethanol

-

30% increase in activity

677787

beta-mercaptoethanol

Agarivorans sp. LQ48

C9WIW7

beta-mercaptoethanol even at a low concentration of 10 mM increases the activity of AgaA by 23%

705482

CaCl2

-

stimulation

dithiothreitol

EDTA

NaCl

additional information

A8W969

no activation of rAgaB34 by chelators (EDTA and EGTA), reducing reagents (beta-mercaptoethanol and DTT), and detergents (SDS and urea)

-

0.0182 - 4.2

agarose

1 - 2

liquid-phase agarose

-

12.81

neoagarodecaose

-

12.39

neoagarododecaose

-

1.7

neoagarohexaose

-

8.36

neoagarotetradecaose

-

additional information

-

22 - 2400

agarose

0.19

neoagarodecaose

-

4.77

neoagarododecaose

-

5.16

neoagarotetradecaose

-

5.3 - 2700

agarose

0.1

KCl

-

0.1

NaCl

-

0.1

NH4Cl

-

0.04

-

culture supernatant, pH 7.0, 30°C

0.1

-

AgaB, 1 ml of 0.125% melted agarose, 44°C, pH 7.5

0.16

-

AgaAc, 1 ml of 0.125% melted agarose, 44°C, pH 7.5

0.7

-

crude enzyme from culture fluid, in 50 mM phosphate buffer, pH 7.0, 35°C

0.872

-

cell lysate

0.9

K7WTF8

cell-free extract, at pH 7.0 and 35°C

732321

1.2

-

wild-type

1237

-

crude enzyme, at pH 6.0 and 50°C

14.2

-

pH 7.0, 40°C

16.4

16.9

culture fluid

1600

-

purified refolded AgaB

167

-

pH 8.0, 40°C

678873

204.4

using agar as substrate, in pH 5.5 acetate buffer, at 45°C

207.5

using agarose as substrate, in pH 5.5 acetate buffer, at 45°C

208.1

K7WTF8

purified enzyme, at pH 7.0 and 35°C

22

-

230.1

-

agarase AG-a

234

-

242.2

15fold purified rAgaB34, with agarose as substrate at 40°C

25.5

Pseudoalteromonas antarctica

-

40°C, pH 8.0

677787

290

-

136884

3

-

cell-free medium, at pH 7.0 and 35°C

329

recombinant AgaC

34

-

36

mutant enzyme V109G/V110C/T111H/S112L, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40°C for 10 min

371

-

50 mM N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid-NaOH buffer, pH 7.8, containing 10 mM CaCl2, 0.1 mM NaCl, 40°C

668031

373.62

-

47.78fold purified enzyme, at pH 8.0 and 40°C

4.13

-

concentrated agarase solution

4.22

-

after 103.5fold purification, pH 7.0, 30°C

43.3

-

136882

466

mutant enzyme N103T, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40°C for 10 min

475

mutant enzyme S182I, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40°C for 10 min

482

wild type enzyme, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40°C for 10 min

5000

recombinant AgaB 476fold purified

517.3

57.45

-

agarase Hz-c, 13.9fold purified

616

-

136871

63.6

-

catalytic module

6447

-

5.21fold purified enzyme, at pH 6.0 and 50°C

7142

-

136885

76.8

-

agarase Hz-b, 18.6fold purified

83.5

84.2

-

after 120.2fold purification, in 50 mM phosphate buffer, pH 7.0, 35°C

9.3

-

5.2

-

porphyran as substrate

136881

5.5

6

9 entries

6 - 7

-

6 - 9

-

6.3

-

6.5

6.5 - 7.5

native enzyme

6.7

Pseudomonas-like bacterium

-

beta-agarase I and IIb

6.8

-

porphyran as substrate

7

7.2

7.4

-

porphyran as substrate

136881

7.5

7.5 - 8

-

Britton-Robinson universal buffers, MOPS and N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid-NaOH buffer

7.6

7.8

8

9

-

additional information

-

mutant L122Q/N446I has comparable optimum pH as the wild-type

4 - 8

4 - 9

-

the enzyme shows more than 90% activity between pH 5.0 and 8.0. More than 50% of the agarase activity is lost below pH 4.0 and above pH 9.0

4.5 - 7.5

the enzyme retains its activity up to 60% at pH 4.5-7.5

5 - 10

-

pH 5.0: about 65% of maximal activity, pH 10.0: about 75% of maximal activity, agarase AG-a

5 - 8

-

pH 5.0: about 35% of maximal activity, about 40% of maximal activity

5 - 8.5

5 - 9

Vibrio harveyi

-

about 60% of activity

136872

5.5 - 7.5

-

the agarase exhibits more than 90% of the maximum activity in the pH range of 5.5 to 7.5

5.5 - 8

-

136880

6 - 10

-

6 - 8

6 - 9

20 - 40

30

34

-

porphyran as substrate

136881

35

37

-

136885

38

Pseudomonas-like bacterium

-

beta-agarase I

38 - 55

native enzyme

40

40 - 41

-

42.5

-

43

Pseudomonas-like bacterium

-

beta-agarase IIb

50

54

-

55

60

-

more than 70% of maximum activity

678873

70

10 - 40

-

the enzyme maintains 85% of the maximum activity at 10°C, the enzyme activity drops at temperatures higher than 40°C

20

-

30% of maximum activity

25 - 40

-

the enzyme works efficiently in the range of 25-40°C and shows maximum activity at 40°C but loses essentially all of its activity at 60°C

25 - 47

more than 50% activity between 25 and 47°C. The enzyme activity drastically decreases over 45°C

25 - 50

-

136885

25 - 55

K7WTF8

the enzyme activity is more than 80% at 25°C and 45°C and less than 10% over 55°C

732321

25 - 70

-

25°C: about 60% of maximal activity, 70°C: about 80% of maximal activity, agarase AG-a

30

-

90% of maximum activity

30 - 45

-

40 - 50

-

the agarase exhibits more than 90% of the maximum activity in the temperature ranging from 40°C to 50°C

40 - 55

more than 80% of the maximum activity remain within 40-55°C

40 - 70

-

at 60 °C, the enzyme activity is 75% of the maximum activity, and enzyme activity declines sharply at 70°C

45 - 85

-

45°C: about 50% of maximal activity, 85°C: about 55% of maximal activity

4.1

calculated

4.14

-

4.2

-

isoelectric focusing, agarase I, principal enzyme

655312

4.7

calculated from amino acid sequence

5

-

calculated from amino acid sequence

5.4

calculated from amino acid sequence

5.9

calculated from amino acid sequence

Acinetobacter sp.

Acinetobacter sp. AG LSL-1

Agarivorans albus YKW-34

-

-

Agarivorans sp. JA-1

Agarivorans sp. JAMB-A11

-

-

-

UniProt

-

-

E-1

-

Alteromonas sp. SY37-12

MK03

656317

-

Bacillus sp. (in: Bacteria) MK03

MK03

-

-

-

-

-

-

Catenovulum agarivorans YM01

UniProt

-

UniProt

Cellvibrio sp. KY-YJ-3

-

-

Cellvibrio sp. OA-2007

-

UniProt

-

136876, 136878, 136879, 136880, 136881, 705484

-

Escherichia coli

-

-

Flammeovirga yaeyamensis

-

Flammeovirga yaeyamensis YT

UniProt

-

UniProt

-

136872, 136877, 705484

-

-

-

-

-

-

Microbulbifer sp. CMC-5 (MTCC 9889)

-

-

Microbulbifer sp. JAMB-A94

-

-

Microbulbifer thermotolerans JAMB-A94

UniProt

-

Pseudoalteromonas antarctica

UniProt

-

136884, 705484

-

Pseudoalteromonas antarctica N-1

-

-

-

136869, 136872, 136874, 136875, 136882, 136883, 697125

-

-

UniProt

-

-

-

-

Pseudomonas-like bacterium

-

-

-

Q21HB9, Q21LJ1

UniProt

Saccharophagus degradans 2-40 / ATCC 43961

Streptomyces coelicolor A3(2)

-

136870, 136871, 136873, 136883, 136885, 713923

-

-

-

-

-

-

UniProt

-

-

-

-

-

UniProt

-

UniProt

-

-

Zobellia galactanivorans Dsij

Dsij

UniProt

-

-

culture fluid

additional information

-

3wz1, download, 3D-view

SCOPe (3wz1)

CATH (3wz1)

100000

105000

106062

x * 106062, calculated, x * 120000, SDS-PAGE of recombinant protein

677750

106100

-

sequence analysis

107000

-

109000

109450

-

x * 109450, calculated, x * 109000, SDS-PAGE

110000

x * 110000, SDS-PAGE

113000

120000

127000

Pseudomonas-like bacterium

-

gel filtration, beta-agarase I

13400

-

AgaAc, calculated by sequence

14000

Pseudomonas-like bacterium

-

alpha2, 2 * 14000, agarase I

140000

Pseudomonas-like bacterium

-

SDS-PAGE, beta-agarase I

146000

Q21HB4, Q21HC5, Q21LJ2

recombinant enzyme

180000

-

gel filtration

136888

20000

21000

-

SDS-PAGE

136872

210000

Pseudomonas-like bacterium

-

analytical ultracentrifugation, beta-agarase I

26000

-

136872, 705484

26500

-

gel filtration

28000

-

SDS-PAGE

136873

29000

-

calculation from sequence of DNA

136870

31000

G0L322, Q9RGX8

recombinant enzyme

32000

32150

-

AgaAc, ESI MS

33000

35130

-

calculation from sequence of an open reading frame

136870

353000

-

AgaB, calculated by sequence

36000

-

SDS-PAGE

37000

recombinant enzyme

39500

40010

-

AgaB, ESI MS

47000

1 * 47000, SDS-PAGE

47200

SDS-PAGE

47240

sequence analysis

48000

48200

-

calculated from sequence

48400

calculated from amino acid sequence

49000

50000

50350

-

x * 50000, SDS-PAGE, x * 50350, calculated, recombinant protein including Huis-tag

677689

50400

1 * 50400, SDS-PAGE

50824

-

x * 50824, calculated

50900

sequence analysis

51000

51200

calculated from amino acid sequence

52000

52580

x * 52580, calculated from amino acid sequence

732321

539000

-

AgaA, calculated by sequence

54000

55000

56000

-

gel filtration

57400

Flammeovirga yaeyamensis

2 * 57400, calculated from amino acid sequence

58000

59000

60000

G0L322, Q9RGX8

recombinant enzyme

61000

Pseudomonas-like bacterium

-

gel filtration, beta-agarase IIb

63000

-

SDS-PAGE

136872

63600

Pseudomonas-like bacterium

-

analytical ultracentrifugation, beta-agarase IIb

64000

65020

-

calculated from sequence

669065

70000

-

x * 70000, SDS-PAGE

71000

-

x * 71000, SDS-PAGE

656476

72000

-

SDS-PAGE

136887

72500

maltose binding protein-fused enzyme, SDS-PAGE

78771

1 * 78771, calculated from amino acid sequence

79000

gel filtration

82000

-

SDS-PAGE

136888

84000

-

His-tagged recombinant enzyme, SDS-PAGE

701877

86000

Q21HB9, Q21LJ1

recombinant enzyme

87000

recombinant enzyme

87500

native enzyme

89000

Q21HB4, Q21HC5, Q21LJ2

recombinant enzyme

92000

98000

-

x * 98000, SDS-PAGE

?

dimer

homodimer

monomer

additional information

glycoprotein

Pseudomonas-like bacterium

-

galactose, N-acetylglucosamine and glucosamine

proteolytic modification

sitting drop vapor diffusion method, using 0.1 M HEPES pH 7.0, 30% PEG 4000, 0.1 M ammonium sulfate and 15% glycerol

-

713635

carbohydrate binding module CBM6-2 of isoform Aga16B in complex with neoagarohexaose. The carbohydrate binding module targets the equatorial O4 and the axial O3 of the anhydro-L-galactose moiety

-

680657

E147S mutant in complex with agaro-octaose, 1.7 A resolution, structure determination by molecular replacment, hanging drop vapor duffusion method, 20.5°C, 30% PEG 4000, 200 mM ammonium acetate, 100 mM sodium acetate

hanging drop vapor diffusion method, using 28-34% (w/v) PEG 8000 and 0.1 M imidazole pH 8.0, at 19°C

Q9RGX8

701557

inactive mutant A-E174S, in complex with agaro-octaose

-

657375

recombinant protein expressed in Escherichia coli

G0L322, Q9RGX8

656176

L122Q

-

shows similar thermostability with wild-type AgaB, raises specific activity 1.3fold

L122Q/N446I

-

has higher thermostability and slightly increased specific activity (1.1fold) than wild-type AgaB. Melting temperature of S2 is 4.6°C higher than that of wild-type AgaB, and the half-life of S2 is 350 min at 40°C, which is 18.4fold longer than that of the wild-type enzyme. Comparable pH stability and optimum pH and temperature profiles as the wild-type

N103T

the mutant shows specific activity similar to the wild type enzyme

N446I

-

has similar enhanced thermostability as mutant L122Q/N446I, and decreased specific activity by 10-20% as compared to the wild-type

N446L

-

the half-life at 40°C is 12.9fold longer than that of wild-type AgaB

N446V

-

the half-life at 40°C is 18.2fold longer than that of wild-type AgaB

S182I

the mutant shows specific activity similar to the wild type enzyme

V109G/V110C/T111H/S112L

the mutant shows severely reduced specific activity

E147S

inactive mutant

E99K

-

the mutant shows 140% relative activity compared to the wild type enzyme

E99K/T307I

-

the mutant shows 200% relative activity compared to the wild type enzyme

T307I

-

the mutant shows 190% relative activity compared to the wild type enzyme

Zobellia galactanivorans Dsij

E147S

-

inactive mutant

-

E99K

-

the mutant shows 140% relative activity compared to the wild type enzyme

-

E99K/T307I

-

the mutant shows 200% relative activity compared to the wild type enzyme

-

T307I

-

the mutant shows 190% relative activity compared to the wild type enzyme

-

additional information

3 - 11

4 - 10

-

4°C, 6 h, stable in the pH range of 4.010.0, agarase AG-a

699362

4 - 8

Vibrio sp. PO-303

A5A6R7

-

705484

4 - 9

4.5 - 8

-

704961

5 - 8

-

705484

5 - 9

5.5 - 11

5.7 - 10.6

6 - 11

6 - 9

6.5 - 8.5

-

the enzyme is stable (more than 70% relative activity) in buffers with a pH range of 6.5-8.5

750214

7 - 8.6

G3CHF0

the enzyme is most stable from pH 7.0 to 8.6

713851

7.1 - 8.2

8

Bacillus subtilis

-

the enzyme is stable at pH 8.0 after 30 min incubation

713704

8 - 9

Q9KK55

-

705484

additional information

-

mutant L122Q/N446I has comparable pH stability as the wild-type

695671

10 - 50

-

up to 80% thermostability of the enzyme is retained at 10°C and 30°C for 180 min. However, when the temperature is increased to 50°C for 30 min, the enzyme relative activity is decreased to 10-20%. The enzyme is fairly stable (almost 70% relative activity) at 40°C for 30 min

100

-

in the presence of 10 mM CaCl2, approximately 17% remaining activity is detected after 30 min

20 - 40

the enzyme is stable from 20 to 40°C after 30 min of incubatiopn. At 40°C, the enzyme retains 90% activity, however, it drastically decreases by 22% of the original value at 42.5°C

20 - 45

-

heat treatment in the range of 20-40°C does not affect the residual activity of the enzyme. Only 45% of the activity is retained after heating at 50°C for 1 h

25

stable up to 25°C

25 - 45

30

30 - 60

-

the enzyme is very stable after incubation for 30 min at 30-50°C, at 60°C 40% of the maximum enzyme activity is remaining

35

37

4 - 30

the enzyme retains its activity when the temperature was between 4 and 30°C for 1 h

40

45

47.5

-

loss in activity 53%

50

50 - 60

50 - 90

-

the agarase is heat-labile, with rapid loss of activity when treated at 50°C for 15 min or at 70°C for 1 min. It loses above 80% of its original activity after incubation at 90°C for 20 s

55

60

65

65 - 80

-

the enzyme retains approximately 90% of the initial activity after incubation for 1 h at 65-80°C

70

-

1 min, 20% residual activity

95

-

the enzyme retains 50% activity after 1 h at 95°C

additional information

-

mutant L122Q/N446I has higher thermostability than wild-type AgaB. Melting temperature of S2 is 4.6°C higher than that of wild-type AgaB, and the half-life of mutant L122Q/N446I is 350 min at 40°C, which is 18.4fold longer than that of the wild-type enzyme. Mutant N446I has similar enhanced thermostability as mutant L122Q/N446I. L122Q shows similar thermostability with wild-type AgaB. Half-life of mutant N446V at 40°C is 18.2fold longer than that of wild-type AgaB. Half-life of mutant N446L at 40°C is 12.9fold longer than that of wild-type AgaB

maintains more than half of its maximum activity at an NaCl concentration of 4 M

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715890

salt requirement for stability

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the agarase shows almost the same activity in both the 0 mM and 1000 mM NaCl solutions, and even

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715890

-20°C

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-20°C, 20 mM Tris-HCl, more than 12 months

Pseudoalteromonas antarctica

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136887

-20°C, a year

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136884

-20°C, at least 6 months

Pseudomonas-like bacterium

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-20°C, more than 2 years

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20°C, two months

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136875

25°C, 2 days

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4°C, 20 mM Tris-HCl buffer, more than 12 months

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136883

4°C, pH 7.5 (phosphate buffer), 6 months, agarase AG-a loses 8% activity

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4°C, Tris-HCl with 0.4 M NaCl, pH 7.5, more than 6 months

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11 entries

17.9fold with a final yield of 23.3%

18.8fold and with a final yield of 16.8% by anion exchange and hydrophobic chromatographies

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476fold and with a final yield of 21.2%, by ammonium sulfate precipitation, weak anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration

agarose bead chromatography

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ammonium sulfate precipitation

Acinetobacter sp.

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682985

ammonium sulfate precipitation and HiTrap DEAE column chromatography

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ammonium sulfate precipitation and Sephadex gel filtration

Bacillus subtilis

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713704

ammonium sulfate precipitation, DEAE-Sepharose column chromatography, and Sephacryl S-100HR gel filtration

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ammonium sulfate precipitation, DEAE-Toyopearl PAK-650M column chromatography, and Sephacryl S-200 gel filtration

ammonium sulfate precipitation, gel filtration on Sephacryl S-100HR, ion exchange chromatography on diethylaminoethyl-Sepharose

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ammonium sulfate precipitation, phenyl Sepharose column chromatography, and Superdex 75 gel filtration

ammonium sulfate precipitation, Q-Sepharose column chromatography, and Sephacryl S-200 gel filtration

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ammonium sulfate precipitation, Sepharose CL6B, anion-exchange Mono Q HR5, Ni-NTA column, AgaB further gel filtration with Sephacryl S200

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ammonium sulfate precipitation, Toyopearl QAE-550C column chromatography, Toyopearl HW-55F column chromatography, and Mono-Q column chromatography

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chitin bead column chromatography

from culture supernatant

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677787

His-trap HP Ni2+ column chromatography and Sephacryl S-200 gel filtration

Flammeovirga yaeyamensis

HisTrap column chromatography

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701877

isolated from the sediment in Suruga bay, Japan, at a depth of 2406m, purification by ammonium sulfate precipitation, DEAE-Toyopearl 650M, gel filtration

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isolated from the sediment of south side of the Kuril trench, Japan, at a depth of 4152 m, purification by ammonium sulfate precipitation, dialysis, DEAE-Toyopearl 650M column, dialysis

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668031

Ni affinity column chromatography and Sephacryl S-200 gel filtration

Q9RGX8

701557

Ni-NTa affinity chromatography and exclusion chromatography with Sephacryl S100

Ni-NTA column chromatography

Ni2+-Sepharose column chromatography

Escherichia coli

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nickel-affinity column chromatography, HiTrap DEAE column chromatography, and Superdex 200 gel filtration

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713635

nickel-chelating affinity column chromatography

partial

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655312

rAgaB34 purified by ultrafiltration and on Ni+ column, 15fold with a yield of 80%

A8W969

recombinant AgaB and mutants purified by ammonium sulfate precipitation, weak anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration. Mutant S2 purified about 476.5fold, with a final yield of 20.9%

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recombinant protein isolated from culture broth

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678898

to electrophoretic purity by immobilized metal affinity chromatography

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to homogeneity directly from the native-PAGE without stained by Coomassie brilliant blue, agarase Hz-b 18.6fold purified and agarase Hz-c 13.9fold purified

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ultrafiltration, DEAE Sepharose column chromatography and Sephacryl S-300HR gel filtration

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using the pMAL protein fusion and purification system

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AgaA, AgaB and truncated form of AgaA referred to as AgaAc with His tag, overexpression in Escherichia coli

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agaB gene inserted into vector pBV220, expressed in Escherichia coli BL21 (DE3) as a fusion protein bearing a C-terminal hexahistidine tag

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DNA fragments (3.0-8.0 kb) ligated into vector pUC18 and transformed into Escherichia coli DH5alpha. Vector pUSH1, which produces a derivative of rAgaB34 with a 6 × His tag at its C-terminal, transformed into Escherichia coli DH5alpha

A8W969

expressed in Bacillus subtilis

expressed in Escherichia coli BL21 cells

expressed in Escherichia coli BL21(DE3) cells

11 entries

expressed in Escherichia coli DH5alpha cells

expressed in Escherichia coli EPI300 cells

expressed in Escherichia coli JM83 cells

Q9KK55

expressed in Pichia pastoris

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expressed in Streptomyces lividans strain TK24

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expression in Bacillu subtilis strain DB104

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678898

expression in Bacillus subtilis

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668031

expression in Bacillus subtilis ISW 1214

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expression in Escherichia coli

expression in Escherichia coli of full-length protein, of the catalytic module, and of the carbohydrate binding module

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expression in Escherichia coli XL21 Blue and ORIGAMI (DE3) pLysS cells

expression in Streptomyces lividans, natural Dag-

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136871

ligated into a pET22b(+) vector and expressed in Escherichia coli BL21(DE3) competent cells

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ligated into pBluescript II KS(+), and transferred into Escherichia coli DH5alpha, forming a genomic DNA library. Cloned into the pET-24a(+) vector and transformed into Escherichia coli BL21 (DE3) competent cells

mutants cloned into vector pET-24a (+) and expressed in Escherichia coli BL21 (DE3). Expression of the agaB gene in vector pBS-ks(sv) and transformed to the Escherichia coli DH5alpha strain

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PCR product cloned into a pT7Blue-2 vector. Amplified DNA fragment digested with NdeI and XhoI and ligated into a pET22b(+) vector and expressed in Escherichia coli BL21(DE3)

the deduced enzyme protein contains a N-terminal signal peptide of 29 amino acid residues, followed by a 266 amino acid sequence that is homologous to catalytic module of family 16 glycoside hydrolases, a bacterial immunoglobulin group 2-like domain of 52 amino acid residues, two carbohydratebinding modules of family 6 separated from Big-2-like domain by nine times repeated GDDTDP amino acid sequence. Expression in Escherichia coli

677750

the enzyme is inducible by agar

using 1 M urea, 500 mM L-arginine, 10% (w/v) glycerol and 20 mM sodium phosphate (pH 7.0), at 4°C

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analysis

industry

synthesis