Part:BBa_K3748004

Secreted Alpha amylase from Bacillus amyloliquefaciens

Alpha amylase is a secreted protein that acts extracellularly, in this part, the native secretion signal has been removed. This enzyme do the catalysis of the endohydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides containing three or more alpha-(1->4)-linked D-glucose units. The product of the reaction are shorter chain saccharides that are then accesible to enter the cell where they can be used as a source of energy. Its native sequence is composed of its signal peptide and the sequence for the active enzyme. This enzyme is naturally secreted in B.amyloliquefaciens.

Alpha amylase has been successfully expressed in S.cerevisiae, using pPGK1 and the secretion signal αMFΔ. Alpha amylase activity has been tested in a colorimetric quantitative assay. If you want to know more go to https://2021.igem.org/Team:Groningen/Experiments.

Figure 1. Increase in absorbance over time as a result of p-nitrophenol generation due to the cleavage of ethylidene-pNP-G7 (polysaccharide) by alpha amylase in the supernatant of S.cerevisiae overnight culture.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 190
  Illegal XbaI site found at 672
  Illegal SpeI site found at 829
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 190
  Illegal SpeI site found at 829
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 190
  Illegal BamHI site found at 116
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 190
  Illegal XbaI site found at 672
  Illegal SpeI site found at 829
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 190
  Illegal XbaI site found at 672
  Illegal SpeI site found at 829
  Illegal AgeI site found at 754
  
• 1000
  COMPATIBLE WITH RFC[1000]