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Literature summary for 3.2.1.135 extracted from

  • Labes, A.; Karlsson, E.N.; Fridjonsson, O.H.; Turner, P.; Hreggvidson, G.O.; Kristjansson, J.K.; Holst, O.; Schoenheit, P.
    Novel members of glycoside hydrolase family 13 derived from environmental DNA (2008), Appl. Environ. Microbiol., 74, 1914-1921.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
Bifidobacterium breve cultured in reinforced clostridial medium, anaeorbic, 37°C. Escherichia coli cultured in Luria Bertani broth, agitated, 37°C. Lactococcus lactis cultured in M17 broth containing 0.5% glucose, 30°C for amplification of vectro pNZ8048 containing only alpha-amylase or pullulanase encoding domains, respectively. Selection of recombinant Escherichia coli with pORI19-apuB on agar containing erythromycin and 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside and isopropyl-beta-D-galactopyranoside Bifidobacterium breve

Protein Variants

Protein Variants Comment Organism
additional information knockout variant through insertion in gene apuB that is encoding for amylopullulanase: no growth on starch, amylopectin, glycogen, or pullulan Bifidobacterium breve

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
175000
-
matured enzyme linked to peptidoglycan at carboxy terminus, premature 182360 Da product of 5127 bp apuB gene with 1708 amino acids, signal sequence of 34 amino acids with cleavage site located between Ala34 and Asp35, 4 highly conserved regions, calculated from sequence Bifidobacterium breve

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
pullulan + H2O Bifidobacterium breve N-terminal alpha-amylase-containing domain and C-terminal pullulanase-containing domain panose + ?
-
?
pullulan + H2O Bifidobacterium breve UCC2003 N-terminal alpha-amylase-containing domain and C-terminal pullulanase-containing domain panose + ?
-
?

Organism

Organism UniProt Comment Textmining
Bifidobacterium breve
-
-
-
Bifidobacterium breve UCC2003
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Lactococcus lactis cytoplasmic fraction containing the alpha-amylase and pullulanase domain proteins, respectively, is purified by using Ni-nitrilotriacetic acid matrices, elution fractions analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis Bifidobacterium breve

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
pullulan + H2O N-terminal alpha-amylase-containing domain and C-terminal pullulanase-containing domain Bifidobacterium breve panose + ?
-
?
pullulan + H2O N-terminal alpha-amylase-containing domain and C-terminal pullulanase-containing domain Bifidobacterium breve UCC2003 panose + ?
-
?

Synonyms

Synonyms Comment Organism
amylopullulanase type II bifunctional amylopullulanase: N-terminal alpha-amylase-containing domain to cleave alpha-1,4-glucosidic linkages such in starch, amylopectin, and glycogen to produce maltooligosaccharides, and a separate C-terminal pullulanase-containing domain to cleave alpha-1,6-glucosidic linkages in pullulan to produce maltotriose and polymers of maltotriose Bifidobacterium breve