Coding

Part:BBa_K3748005

Designed by: Sofía Jiménez Ochoa   Group: iGEM21_Groningen   (2021-10-01)


Secreted Alpha amylase from Bacillus licheniformis

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.licheniformis.

Alpha amylase has been successfully expressed in S.cerevisiae, using pRNR2 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.

T--Groningen--Parts_3bBL.png

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
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None