Part:BBa_K1356000
Alpha Amylase with TolB Signaling Sequence and PstI Site Removed
The alpha amylase for this part was taken from part BBa_K1195001. Attached to it is a DsbA signaling sequence required in order to express the gene product extracellularly in N. multiformis in the break down of biofilm in wastewater treatment plants. Additionally, the PstI site originally found in the BBa_K1195001 part was removed using site-directed mutagenesis. The restriction site was changed from "CTGCAG" to "CTCCAG" in order to remove this site. This gene is contained within the standard iGem pSB1C3 plasmid backbone.
Amylase is an enzyme naturally synthesized by bacteria, such as E. coli, fungi, and even in humans in saliva and the pancreas. This enzyme catalyzes the hydrolysis of starches into sugars and breaks down the components of bacterial extracellular polymeric substance (EPS), which contains extracellular DNA, polysaccharides, and proteins. These components are a primary part of most bacterial biofilms and it is hoped that the enzyme being expressed extracellularly will allow for more biofilm break down so that N. multiformis can more effectively breakdown wastewater components to make wastewater treatment plants more effective. It has been shown in other studies that amylase is an effective degrader of several other types of biofilms and we hope to show that it is equally effective at breaking down wastewater biofilm.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Plasmid Schematic
RaptorX Protein Structure and Function Prediction
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