Part:BBa_K3748006

Secreted Alpha amylase from Bacillus subtilis

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.subtilis.

Alpha amylase has been successfully expressed in S.cerevisiae, using pPGK1 and the secretion signal αMF. 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 607
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 607
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 607
  Illegal BglII site found at 1079
  Illegal BglII site found at 1249
  Illegal BamHI site found at 1099
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 607
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 607
  
• 1000
  COMPATIBLE WITH RFC[1000]