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Literature summary for 2.4.1.18 extracted from

  • Suzuki, R.; Koide, K.; Hayashi, M.; Suzuki, T.; Sawada, T.; Ohdan, T.; Takahashi, H.; Nakamura, Y.; Fujita, N.; Suzuki, E.
    Functional characterization of three (GH13) branching enzymes involved in cyanobacterial starch biosynthesis from Cyanobacterium sp. NBRC 102756 (2015), Biochim. Biophys. Acta, 1854, 476-484.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene nbrc_glgB1, functional recombinant expression of N-terminally His-tagged isozyme BE1 in Escherichia coli Cyanobacterium sp.
gene nbrc_glgB2, functional recombinant expression of His-tagged isozyme BE2 in Escherichia coli Cyanobacterium sp.
gene nbrc_glgB3, functional recombinant expression of His-tagged isozyme BE3 in Escherichia coli Cyanobacterium sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
66000
-
x * 77262, sequence calculation of His-tagged isozyme BE3, x * 66000, recombinant His-tagged isozyme BE3, SDS-PAGE Cyanobacterium sp.
69000
-
x * 78781, sequence calculation of His-tagged isozyme BE2, x * 69000, recombinant His-tagged isozyme BE2, SDS-PAGE Cyanobacterium sp.
77262
-
x * 77262, sequence calculation of His-tagged isozyme BE3, x * 66000, recombinant His-tagged isozyme BE3, SDS-PAGE Cyanobacterium sp.
78781
-
x * 78781, sequence calculation of His-tagged isozyme BE2, x * 69000, recombinant His-tagged isozyme BE2, SDS-PAGE Cyanobacterium sp.
88000
-
x * 94125, sequence calculation of His-tagged isozyme BE1, x * 88000, recombinant His-tagged isozyme BE1, SDS-PAGE Cyanobacterium sp.
94125
-
x * 94125, sequence calculation of His-tagged isozyme BE1, x * 88000, recombinant His-tagged isozyme BE1, SDS-PAGE Cyanobacterium sp.

Organism

Organism UniProt Comment Textmining
Cyanobacterium sp. A4UU34 gene nbrc_glgB1, isozyme BE1
-
Cyanobacterium sp. A4UU35 gene nbrc_glgB2, isozyme BE2
-
Cyanobacterium sp. A4UU36 nbrc_glgB3, isozyme BE3
-
Cyanobacterium sp. NBRC 102756 A4UU34 gene nbrc_glgB1, isozyme BE1
-
Cyanobacterium sp. NBRC 102756 A4UU35 gene nbrc_glgB2, isozyme BE2
-
Cyanobacterium sp. NBRC 102756 A4UU36 nbrc_glgB3, isozyme BE3
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged isozyme BE1 from Escherichia coli by nickel affinity chromatography, gel filtration, and ultrafiltration Cyanobacterium sp.
recombinant His-tagged isozyme BE2 from Escherichia coli by nickel affinity chromatography, gel filtration, and ultrafiltration Cyanobacterium sp.
recombinant His-tagged isozyme BE3 from Escherichia coli by nickel affinity chromatography, gel filtration, and ultrafiltration Cyanobacterium sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
47.3
-
purified recombinant His-tagged isozyme BE1, pH 7.0, 30°C Cyanobacterium sp.
951.4
-
purified recombinant His-tagged isozyme BE1, pH 7.0, 30°C Cyanobacterium sp.
988.8
-
purified recombinant His-tagged isozyme BE1, pH 7.0, 30°C Cyanobacterium sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the recombinant isozyme is active with amylose or amylopectin, and is active against phosphorylase limit dextrin, in which outer branches are uniformly diminished to 4 glucose residues. Chain-length preference of reaction product of the recombinant isozymes, overview Cyanobacterium sp. ?
-
?
additional information the recombinant isozyme is active with amylose or amylopectin, and is active against phosphorylase limit dextrin, in which outer branches are uniformly diminished to 4 glucose residues. Chain-length preference of reaction product of the recombinant isozymes, overview Cyanobacterium sp. NBRC 102756 ?
-
?

Subunits

Subunits Comment Organism
? x * 77262, sequence calculation of His-tagged isozyme BE3, x * 66000, recombinant His-tagged isozyme BE3, SDS-PAGE Cyanobacterium sp.
? x * 78781, sequence calculation of His-tagged isozyme BE2, x * 69000, recombinant His-tagged isozyme BE2, SDS-PAGE Cyanobacterium sp.
? x * 94125, sequence calculation of His-tagged isozyme BE1, x * 88000, recombinant His-tagged isozyme BE1, SDS-PAGE Cyanobacterium sp.
More branching enzymes are multidomain proteins, GH13-type BEs contain (from N- to C-terminal end) carbohydrate binding module 48 (CBM48), catalytic domain (i.e. domain A), and domain C, isozyme domain structure comparisons, overview Cyanobacterium sp.

Synonyms

Synonyms Comment Organism
BE1
-
Cyanobacterium sp.
BE2
-
Cyanobacterium sp.
BE3
-
Cyanobacterium sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Cyanobacterium sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35
-
purified recombinant His-tagged isozyme BE3, stable up to Cyanobacterium sp.
40
-
purified recombinant His-tagged isozyme BE1, stable up to Cyanobacterium sp.
45
-
purified recombinant His-tagged isozyme BE2, stable up to Cyanobacterium sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Cyanobacterium sp.
8
-
-
Cyanobacterium sp.

pH Range

pH Minimum pH Maximum Comment Organism
8 10 isozyme BE2 shows nearly half of the maximum activity at pH 10.0 Cyanobacterium sp.

General Information

General Information Comment Organism
evolution Cyanobacterium sp. NBRC 102756 belongs to the cyanobacteria that accumulate water-insoluble alpha-glucan similar to amylopectin rather than glycogen, the latter of which is more commonly produced in these organisms. The amylopectin-producing species invariably have three branching enzyme (BE) homologues, BE1, BE2, and BE3, all belonging to the glycoside hydrolase family 13, GH13, phylogenetic tree. The specific activity of BE3 is much lower than those of BE1 and BE2. Isozymes BE1 and BE2 show similar chain-length preference to BEIIb isoform of Oryza sativa, while the catalytic specificity of BE3 is similar to that of Oryza sativa BEI. These results indicate that starch-producing cyanobacteria have both type-I BE (BE3) and type-II BEs (BE1 and BE2) in terms of chain-length preferences, as is the case of plants Cyanobacterium sp.
evolution Cyanobacterium sp. NBRC 102756 belongs to the cyanobacteria that accumulate water-insoluble alpha-glucan similar to amylopectin rather than glycogen, the latter ofwhich is more commonly produced in these organisms. The amylopectin-producing species invariably have three branching enzyme (BE) homologues, BE1, BE2, and BE3, all belonging to the glycoside hydrolase family 13, GH13, phylogenetic tree. The specific activity of BE3 is much lower than those of BE1 and BE2. Isozymes BE1 and BE2 show similar chain-length preference to BEIIb isoform of Oryza sativa, while the catalytic specificity of BE3 is similar to that of Oryza sativa BEI. These results indicate that starch-producing cyanobacteria have both type-I BE (BE3) and type-II BEs (BE1 and BE2) in terms of chain-length preferences, as is the case of plants Cyanobacterium sp.