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Literature summary for 3.2.1.78 extracted from

  • Xia, W.; Lu, H.; Xia, M.; Cui, Y.; Bai, Y.; Qian, L.; Shi, P.; Luo, H.; Yao, B.
    A novel glycoside hydrolase family 113 endo-beta-1,4-mannanase from Alicyclobacillus sp. strain A4 and insight into the substrate recognition and catalytic mechanism of this family (2016), Appl. Environ. Microbiol., 82, 2718-2727 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.78

Activating Compound

Activating Compound Comment Organism Structure
2-mercaptoethanol moderate activation at 1 mM Alicyclobacillus sp. A4

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Alicyclobacillus sp. A4

Protein Variants

Protein Variants Comment Organism
C143A the mutant shows reduced activity compared to the wild type enzyme Alicyclobacillus sp. A4
N90A the mutant shows reduced activity compared to the wild type enzyme Alicyclobacillus sp. A4
R97A inactive Alicyclobacillus sp. A4
W13A inactive Alicyclobacillus sp. A4
W13F the mutant shows reduced activity compared to the wild type enzyme Alicyclobacillus sp. A4
W274A inactive Alicyclobacillus sp. A4
W96A inactive Alicyclobacillus sp. A4
Y196A inactive Alicyclobacillus sp. A4
Y292A the mutant shows increased activity with mannotriose and mannotetraose and about 50% reduced activity with mannopentaose and mannohexaose compared to the wild type enzyme Alicyclobacillus sp. A4

Inhibitors

Inhibitors Comment Organism Structure
Ag+ strong inhibition at 1 mM Alicyclobacillus sp. A4
Cu2+ strong inhibition at 1 mM Alicyclobacillus sp. A4
SDS strong inhibition at 1 mM Alicyclobacillus sp. A4

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ moderate activation at 1 mM Alicyclobacillus sp. A4

Organism

Organism UniProt Comment Textmining
Alicyclobacillus sp. A4 W5R5D2
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein
-
 Alicyclobacillus sp. A4

Purification (Commentary)

Purification (Comment) Organism
Ni2+-NTA column chromatography Alicyclobacillus sp. A4

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
galactomannan + H2O
-
 Alicyclobacillus sp. A4 ?
-
 ?
glucomannan + H2O high activity Alicyclobacillus sp. A4 mannobiose + mannotriose
-
 ?
guar gum + H2O
-
 Alicyclobacillus sp. A4 mannose + mannobiose + mannotriose
-
 ?
guar gum + H2O highest activity with a mannose to galactose ratio of 79:21 Alicyclobacillus sp. A4 ?
-
 ?
ivory nut mannan + H2O moderate activity Alicyclobacillus sp. A4 ?
-
 ?
konjac flour + H2O high activity Alicyclobacillus sp. A4 ?
-
 ?
locust bean gum + H2O
-
 Alicyclobacillus sp. A4 ?
-
 ?
mannohexaose + H2O
-
 Alicyclobacillus sp. A4 ?
-
 ?
mannopentaose + H2O
-
 Alicyclobacillus sp. A4 ?
-
 ?
mannotetraose + H2O
-
 Alicyclobacillus sp. A4 mannobiose + mannotriose + mannopentaose
-
 ?
additional information no activity with mannotriose, barley beta-glucan, birchwood xylan, and carboxymethyl cellulose Alicyclobacillus sp. A4 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 39000, SDS-PAGE Alicyclobacillus sp. A4
? x * 39200, calculated from amino acid sequence Alicyclobacillus sp. A4

Synonyms

Synonyms Comment Organism
endo-beta-1,4-mannanase
-
 Alicyclobacillus sp. A4
Man113A
-
 Alicyclobacillus sp. A4

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
 Alicyclobacillus sp. A4

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 60 more than 50% activity between 40 and 60°C Alicyclobacillus sp. A4

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40 60 the enzyme maintains thermostability with 80% activity remaining at 40°C after 1 h and 60% activity remaining at 50°C after 1 h. When incubated at 60°C without substrate, the enzyme loses activity rapidly (about 25% residual activity after 10 min, no activity after 30 min) Alicyclobacillus sp. A4

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
 Alicyclobacillus sp. A4

pH Range

pH Minimum pH Maximum Comment Organism
5 8 more than 50% activity between pH 5.0 and 8.0 Alicyclobacillus sp. A4

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 10 the enzyme is stable over a pH range from 5.0 to 10.0 for 1 h, since it retains more than 60% of the initial activity after incubation without substrate at 37°C for 1 h Alicyclobacillus sp. A4

pI Value

Organism Comment pI Value Maximum pI Value
Alicyclobacillus sp. A4 calculated from amino acid sequence
-
 5.2

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.48
-
 Mannotriose mutant enzyme C143A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
1.3
-
 Mannotriose mutant enzyme N90A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
3.54
-
 Mannotriose mutant enzyme W13F, at pH 6.0 and 50°C Alicyclobacillus sp. A4
6.54
-
 Mannotriose wild type enzyme, at pH 6.0 and 50°C Alicyclobacillus sp. A4
6.98
-
 Mannotetraose mutant enzyme C143A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
7.6
-
 Mannotriose mutant enzyme Y292A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
11.37
-
 Mannotetraose mutant enzyme N90A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
14.8
-
 Mannotetraose mutant enzyme W13F, at pH 6.0 and 50°C Alicyclobacillus sp. A4
16.49
-
 mannohexaose mutant enzyme C143A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
20.34
-
 Mannopentaose mutant enzyme N90A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
21.21
-
 mannohexaose mutant enzyme Y292A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
29.58
-
 Mannopentaose mutant enzyme C143A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
29.7
-
 Mannotetraose wild type enzyme, at pH 6.0 and 50°C Alicyclobacillus sp. A4
30.36
-
 Mannopentaose mutant enzyme Y292A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
31.42
-
 mannohexaose mutant enzyme N90A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
45.63
-
 Mannotetraose mutant enzyme Y292A, at pH 6.0 and 50°C Alicyclobacillus sp. A4
54.48
-
 Mannopentaose mutant enzyme W13F, at pH 6.0 and 50°C Alicyclobacillus sp. A4
72.7
-
 Mannopentaose wild type enzyme, at pH 6.0 and 50°C Alicyclobacillus sp. A4
73.76
-
 mannohexaose mutant enzyme W13F, at pH 6.0 and 50°C Alicyclobacillus sp. A4
75.7
-
 Mannotetraose wild type enzyme, at pH 6.0 and 50°C Alicyclobacillus sp. A4