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Literature summary for 3.2.1.68 extracted from

  • Li, Y.; Niu, D.; Zhang, L.; Wang, Z.; Shi, G.
    Purification, characterization and cloning of a thermotolerant isoamylase produced from Bacillus sp. CICIM 304 (2013), J. Ind. Microbiol. Biotechnol., 40, 437-446.
    View publication on PubMed
Search for more literature for 3.2.1.68

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, phylogenetic tree Bacillus sp. (in: Bacteria)

Inhibitors

Inhibitors Comment Organism Structure
Cu2+ complete inhibition at 0.5 mM Bacillus sp. (in: Bacteria)
Fe3+ 59% inhibition at 0.5 mM, 90% at 5 mM Bacillus sp. (in: Bacteria)
Hg2+ complete inhibition at 0.5 mM Bacillus sp. (in: Bacteria)
additional information no inhibition by EDTA, citrate, maltose, and alpha-cyclodextrin Bacillus sp. (in: Bacteria)
phenylmethanesulfonyl fluoride 9% inhibition at 5 mM Bacillus sp. (in: Bacteria)
SDS 77% inhibition at 30 mM Bacillus sp. (in: Bacteria)
Sn2+ 28% inhibition at 0.5 mM, 73% at 5 mM Bacillus sp. (in: Bacteria)
Urea 82% inhibition at 2 M Bacillus sp. (in: Bacteria)
Zn2+ 64% inhibition at 0.5 mM, 88% at 5 mM Bacillus sp. (in: Bacteria)

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information
-
 Bacillus sp. (in: Bacteria)

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activates 10% at 0.5 mM, and 25% at 5 mM, Ca2+ highly stabilizes the enzyme at 80°C Bacillus sp. (in: Bacteria)
Co2+ activates 12% at 5 mM Bacillus sp. (in: Bacteria)
Mn2+ activates 28% at 0.5 mM, and 41% at 5 mM Bacillus sp. (in: Bacteria)
additional information poor effects by Na+, K+, Mg2+, Li+, and EDTA at 0.5-5 mM. Mn2+ and Mg2+ do not stabilize the enzyme at 70°C and 80°C Bacillus sp. (in: Bacteria)

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
100000
-
 gel filtration Bacillus sp. (in: Bacteria)
101155
-
 1 * 100000, SDS-PAGE, 1 * 101155, sequence calculation Bacillus sp. (in: Bacteria)

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
amylopectin + H2O Bacillus sp. (in: Bacteria)
-
 amylose + linear maltooligosaccharides
-
 ?
amylopectin + H2O Bacillus sp. (in: Bacteria) CICIM 304
-
 amylose + linear maltooligosaccharides
-
 ?

Organism

Organism UniProt Comment Textmining
Bacillus sp. (in: Bacteria) L0C9D3 isolated by screening a soil sample collected from a hot spring of Tengchong volcano in Yunnan, China
-
Bacillus sp. (in: Bacteria) CICIM 304 L0C9D3 isolated by screening a soil sample collected from a hot spring of Tengchong volcano in Yunnan, China
-

Purification (Commentary)

Purification (Comment) Organism
native extracellular enzyme 369fold from culture supernatant by ultrafiltration, ammonium sulfate fractionation, gel filtration, two different steps of anion exchange chromatography, and dialysis, to homogeneity Bacillus sp. (in: Bacteria)

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 6535 U/mg, measuring 0.01 increase in absorbance at 600 nm per h at 50°C, pH 6.0, per mg protein Bacillus sp. (in: Bacteria)

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
amylopectin + H2O
-
 Bacillus sp. (in: Bacteria) amylose + linear maltooligosaccharides
-
 ?
amylopectin + H2O
-
 Bacillus sp. (in: Bacteria) CICIM 304 amylose + linear maltooligosaccharides
-
 ?
glycogen + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
glycogen + H2O
-
 Bacillus sp. (in: Bacteria) CICIM 304 ?
-
 ?
maize starch + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
maltodextrin + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
maltodextrin + H2O
-
 Bacillus sp. (in: Bacteria) CICIM 304 ?
-
 ?
additional information pullulan is a poor substrate Bacillus sp. (in: Bacteria) ?
-
 ?
additional information pullulan is a poor substrate Bacillus sp. (in: Bacteria) CICIM 304 ?
-
 ?
soluble starch + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
soluble starch + H2O
-
 Bacillus sp. (in: Bacteria) CICIM 304 ?
-
 ?
sticky rice starch + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 100000, SDS-PAGE, 1 * 101155, sequence calculation Bacillus sp. (in: Bacteria)

Synonyms

Synonyms Comment Organism
glycogen-6-glucanogydrolase
-
 Bacillus sp. (in: Bacteria)
IAM
-
 Bacillus sp. (in: Bacteria)

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 recombinant enzyme Bacillus sp. (in: Bacteria)

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 70 purified native enzyme, pH 6.0, 1 h, over 70% activity remaining Bacillus sp. (in: Bacteria)
80
-
 purified native enzyme, pH 6.0, 1 h, inactivation Bacillus sp. (in: Bacteria)

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 broad optimum, profile overview Bacillus sp. (in: Bacteria)

pH Range

pH Minimum pH Maximum Comment Organism
5.5 9 over 90% of maximal activity within this range, profile overview Bacillus sp. (in: Bacteria)

pH Stability

pH Stability pH Stability Maximum Comment Organism
4.5 9 purified native enzyme, 50°C, 1 h, over 70% activity remaining Bacillus sp. (in: Bacteria)

General Information

General Information Comment Organism
evolution the enzyme harbors a carbohydrate-binding module family 48 (CBM48) domain Bacillus sp. (in: Bacteria)
physiological function isoamylase catalyzes hydrolysis of alpha-D-(1,6)-glucosidic branch linkages in amylopectin and glycogen to release amylose and linear maltooligosaccharides Bacillus sp. (in: Bacteria)