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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Bifidobacterium breve | - |
- |
- |
Bifidobacterium breve UCC2003 | - |
- |
- |
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 | 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 | 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 |