Information on EC 3.2.1.158 - alpha-agarase

-

agaraseA33

agaW1540

A0A8F2W760

-

AgaWS5

alpha-agarase

CaLJ96

-

769809, 772453

Cm-AGA

Endohydrolysis of (1->3)-alpha-L-galactosidic linkages in agarose, yielding agarotetraose as the major product

-

agarose 3-glycanohydrolase

Requires Ca2+. The enzyme from Thalassomonas sp. can use agarose, agarohexaose and neoagarohexaose as substrate. The products of agarohexaose hydrolysis are dimers and tetramers, with agarotetraose being the predominant product, whereas hydrolysis of neoagarohexaose gives rise to two types of trimer. While the enzyme can also hydrolyse the highly sulfated agarose porphyran very efficiently, it cannot hydrolyse the related compounds kappa-carrageenan (see EC 3.2.1.83) and iota-carrageenan (see EC 3.2.1.157) [2]. See also EC 3.2.1.81, beta-agarase.

63952-00-1

-

agar + H2O

?

agar + H2O

agarotetraose

Substrates: -
Products: -

?

agarohexaose + H2O

?

-

Substrates: -
Products: -

?

agarohexaose + H2O

agarobiose + agarotetraose

agarohexaose + H2O

agarotetraose + agarobiose

agarooctaose + H2O

2 agarotetraose

769890

Substrates: -
Products: -

?

agaropentaose + H2O

agarobiose + agarotriose

-

Substrates: -
Products: -

?

agarose + H2O

?

agarose + H2O

agarobiose + agarotetraose + ?

agarose + H2O

agarobiose + agarotetraose + agarohexaose

-

Substrates: agarotetraose is the dominant product. No actiovity with: agarobiose, agarotriose and agarotetraose
Products: -

?

agarose + H2O

agarotetraose + ?

20 entries

agarose + H2O

agarotetraose + agarobiose + agarohexaose

agarose + H2O

agarotetraose + agarohexaose

agarose + H2O

agarotetraose + agarohexaose + ?

agarose + H2O

neoagarotetraose + neoagarohexaose + ?

A0A8F2W760

Substrates: rAgaW1540 is an endo-acting beta-agarase that degrades agarose into neoagarotetraose and neoagarohexaose as the main products. cf. EC 3.2.1.81
Products: -

?

agarotetraose + H2O

neoagarotriose

Substrates: -
Products: -

?

neoagarohexaose + H2O

?

-

Substrates: -
Products: -

?

neoagarohexaose + H2O

agarotetraose

porphyran + H2O

?

additional information

?

-

agar + H2O

?

agarose + H2O

agarotetraose + ?

Ag+

A0A8F2W760

10 mM, slightly promotes the enzymatic activity

Ba2+

Ca2+

Cr2+

A0A8F2W760

10 mM, slightly promotes the enzymatic activity

Fe2+

A0A8F2W760

10 mM, activation to 126.99% of control

Ga3+

A0A8F2W760

10 mM, slightly promotes the enzymatic activity

K+

A0A8F2W760

10 mM, slightly promotes the enzymatic activity

Li+

Mn2+

Na+

Ni2+

A0A8F2W760

10 mM, slightly promotes the enzymatic activity

Sr2+

A0A8F2W760

10 mM, slightly promotes the enzymatic activity

Ag+

-

1 mM, inhibition to 15% of control

Al3+

3% residual activity at 2 mM

Ba2+

BaCl2

2 mM, 5% loss of activity

Co2+

Cr3+

-

10 mM, inhibition to 32.71% of control

Cu2+

6 entries

CuCl2

2 mM, 48% loss of activity

EDTA

Fe2+

Fe3+

K+

-

10 mM, inhibition to 58.4% of control

Li+

-

5 mM, inhibition to 61.71% of control

LiCl

2 mM, 3% loss of activity

Mg2+

MgCl2

2 mM, 5% loss of activity

Mn2+

MnCl2

2 mM, 43% loss of activity

Na+

-

10 mM, inhibition to 58.1% of control

Na2EDTA

-

10 mM, complete inhibition

NH4+

84% residual activity at 2 mM

Ni2+

5 entries

NiCl2

2 mM, 51% loss of activity

Pb2+

-

1 mM, inhibition to 7% of control

SDS

5 entries

Sn2+

-

1 mM, inhibition to 30% of control

Zn2+

7 entries

Ca2+

-

664530

EDTA

A0A8F2W760

10 mM, slightly promotes the enzymatic activity

additional information

W7Q7M2

the enzyme exhibits a cold activation, which is enhanced by 5fold after incubation at -20°C or 4°C for 6 h. The carbohydrate-binding modules and long linker region are presumed to play a crucial role in cold adaptation due to a relatively high percentage of acidic residues and a negative charge density compared to the catalytic domain

771199

-

additional information

agarose

18 entries

2.4 - 576.96

agarose

6 entries

additional information

agarose

11 entries

206.1

MH005820.1

pH and temperature not specified in the publication

40.7

6423

-

pH 7.2, 42°C

664530

additional information

6

-

7

7.2

7.4

porphyran cleavage assay at

8

8.5

3 - 10

A0A8F2W760

pH 3.0: about 60% of maximal activity, pH 10.0: about 70% of maximal activity

3 - 5

-

the activity is less than 10% of that at pH 7.0

4 - 9

-

pH 4.0: about 70% of maximal activity, pH 9.0: about 40% of maximal activity

4.5 - 9.5

5 - 7

-

pH 5.0: about 55% of maximal activity, pH 7.0: about 75% of maximal activity

6 - 10

pH 6.0: about 70% of maximal activity, pH 10.0: about 70% of maximal activity

768774

6 - 11

-

6 - 9

-

664808

7 - 10

-

more than 80% of the highest enzyme activity is retained at pH 7.0-10.0

7 - 9

MH005820.1

pH 7.0: about 60% of maximal activity, pH 9.0: about 40% of maximal activity

22

cultivation of Eschericha coli cells harbouring the agaA gene, agarose lysis observed after one week on Zd agar broth

35

5 entries

37

-

40

45

0 - 80

A0A8F2W760

0°C: 85.4% of maximal activity, 20-40°C: more than 92% of maximal activity, 80°C: about 70% of maximal activity

10 - 60

10°C: about 40% of maximal activity, 60°C: about 70% of maximal activity

768774

20 - 40

20 - 50

30 - 45

more than 50% of the enzyme activity is lost below 30°C and above 45°C

30 - 50

-

more than 80% of the highest enzyme activity is maintained at 30-50°C, and the activity at 20°C is 12.3% of that at 37°C

5.3

-

isoelectric focusing

664808

-

-

Agarilytica rhodophyticola MCCC 1H00123

-

-

Alteromonas agarilytica GJ1B

Catenovulum agarivorans DS-2

STB13

771199

W7Q7M2

UniProt

Catenovulum agarivorans YM01T

-

732006

-

-

MH005820.1

GenBank

Catenovulum maritimum STB14

-

MH005820.1

GenBank

-

768774, 769809

UniProt

Catenovulum sediminis WS1-A

-

UniProt

-

-

-

A0A8F2W760

UniProt

Thalassomonas sp. JAMB A33

-

664696, 731519, 769809

-

-

731519

-

Thalassomonas sp. JAMB-A33

-

664696, 769809

-

-

680291, 705484

Thalassotalea agarivorans JAMB-A33

UniProt

-

UniProt

additional information

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

140000

-

769809

150000

SDS-PAGE

768774

154000

deduced from amino acid sequence of pre-protein of 1429 residues

155000

-

769809

156090

MH005820.1

calculated

158000

-

180000

200000

360000

50000

A0A8F2W760

SDS-PAGE

54000

-

SDS-PAGE

75000

-

x * 75000, SDS-PAGE

731519

85000

86780

deduced from amino acid sequence

680291

87000

recombinant enzyme

705484

89800

-

calculated

90000

-

SDS-PAGE

97000

SDS-PAGE

?

dimer

homodimer

monomer

additional information

includes the peptides A and B, signal peptide of 26 residues, cleaved between A26 and E27

D259N

-

KM-value for agarose is lower than that of wild-type enzyme

D333N

-

mutant enzyme exhibits no enzymatic activity

E452Q

-

KM-value for agarose is lower than that of wild-type enzyme

E468Q

-

mutant enzyme exhibits no enzymatic activity

D259N

Agarilytica rhodophyticola MCCC 1H00123

-

KM-value for agarose is lower than that of wild-type enzyme

-

D333N

Agarilytica rhodophyticola MCCC 1H00123

-

mutant enzyme exhibits no enzymatic activity

-

E452Q

Agarilytica rhodophyticola MCCC 1H00123

-

KM-value for agarose is lower than that of wild-type enzyme

-

E468Q

Agarilytica rhodophyticola MCCC 1H00123

-

mutant enzyme exhibits no enzymatic activity

-

D1328G

kcat/Km for agarose is 1.9fold lower than the wild-type value

D1330G

kcat/Km for agarose is 1.04fold lower than the wild-type value

D460G

the mutant enzyme retains more than 80% of activity compared with that of the wild-type enzyme

D695G

the mutant enzyme retains more than 80% of activity compared with that of the wild-type enzyme

D708G

the mutant enzyme retains more than 80% of activity compared with that of the wild-type enzyme

D758G

the mutant enzyme retains more than 80% of activity compared with that of the wild-type enzyme

D779G

mutation leads to complete inactivation

D781G

70% loss of activity. D781 is involved in the binding of Na+, and its mutation raises the optimal concentration of Na+ 4 times higher than that of the wild type

additional information

10

11

MH005820.1

6 h, enzyme retains about 45% of its initial activity

3

4

5

5 - 6

-

2 h, enzyme retains 60% of its initial activity

6

6 - 11

6.5

6.5 - 10.5

7

7 - 9

MH005820.1

6 h, enzyme retains about 70% of its initial activity

8

9

0

-

2 h, enzyme retains 90% of its initial activity

10

-

2 h, enzyme retains 100% of its initial activity

10 - 40

the enzyme retains 100% or 60% activity after being incubated at 10-20°C or 30°C for 1 h, respectively. The enzyme loses nearly 95% activity after being incubated at 40°C for 1 h

20

20 - 30

-

1 h, stable

25

A0A8F2W760

60 min, enzyme retains 97.9% of its initial activity

30

35

MH005820.1

1 h, enzyme retains about 40% of its initial activity

37

-

1 h, complete loss of activity at temperature higher than 37°C

40

45

50

60

-

30 min, enzyme retains about 20% of its initial activity

Ca2+ stabilizes enzyme

-

4°C, 28 d, more than 70% of initial activity is retained

-

-

11 entries

AgaE is purified by two steps using ultrafiltration and Ni2+ based metal ion affinity chromatography with the yield of about 5.4 mg/l of recombinant protein

gel filtration

680291

hydroxyapatite chromatography and gel filtration

705484

isolated from the sediment of Noma point, Japan, at a depth of 230m, purified by DEAE-Toyopearl 650M column, ultracentrifugation and gel filtration

-

Ni-Sepharose column chromatography and Superdex S200 gel filtration

-

-

agaA gene isolated from a genomic library of Alteromonas agarilytica, alpha-agarase precursor of 1429 amino acid residues, expressed in Escherichia coli, novel catalytic domain defines a new GH family

expressed in Bacillus subtilis

705484

expressed in Escherichia coli AD494(DE3) cells

expressed in Escherichia coli HB101, open reading frame of 1463 amino acid residues of the agaA33 gene determined, for extracellular production of recombinant alpha-agarase AgaA33 the Bacillus subtilis strain ISW1214 serves as host and plasmid pEXBS as expression vector

680291

expressed in Saccharomyces cerevisiae strains SEY2102 and FY833 and in Pichia pastoris strain GS115

-

731519

expression in Bacillus subtilis

W7Q7M2

771199

expression in Escherichia coli

expression in Escherichia coli BL21 (DE3)

MH005820.1

expression in Escherichia coli BL21(DE3)

-

expression in Escherichia coli BL21(DE3) and in Escherichia coli Rosetta (DE3)

-

expression in Escherichia coli BL21(DE3) plysS

A0A8F2W760

expression in Escherichia coli ER2566