Part:BBa_K3748000

Secreted Alpha Amylase from Bacillus Amyloliquefaciens

Alpha amylase is a secreted protein that acts extracellularly. 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 compose of its signal peptide and the sequence for the active enzyme. This enzyme is naturally secreted in B.amyloliquefaciens.

Alpha amylase has been successfully secreted in S.paradoxus, using pHHF1. 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.paradoxus overnight culture.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 304
  Illegal XbaI site found at 786
  Illegal SpeI site found at 943
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 304
  Illegal SpeI site found at 943
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 304
  Illegal BamHI site found at 230
  Illegal BamHI site found at 1567
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 304
  Illegal XbaI site found at 786
  Illegal SpeI site found at 943
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 304
  Illegal XbaI site found at 786
  Illegal SpeI site found at 943
  Illegal AgeI site found at 868
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 14
  Illegal BsaI.rc site found at 1574