Part:BBa_K1606007

Kumamax enzyme that gluten updated m-cherry reporter device

We modernized m-cherry (BBa_J06504) part to report the absence of the gluten protein in a solution or in a dough that our updated Kumamax enzyme is inactive in the environment. When the absence of the protein of the gluten our repoter gene is expressed and give red light to environment. Besides LacI promoter works under acidic environment in the dough formation.

mCherry(restricted)-GFP-GFP-PGap-z

101-106-117-mcherry-pgap-pgap-pgap-pgap-mcherry

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Project Summary

People with celiac disease have to consume gluten free food because gluten creates an auto-immune response in their body. However, gluten free food is a lot more expensive than normal food. That is why, we decided to find a solution to this problem by taking kumamolisin and putting it in a vector of yeast that has alpha secretion factor then putting the plasmid in yeast. The yeast will produce and secrete kumamolisin enzyme to its external environment and will destroys gluten while the bread continues to be fermented. We also prepared a gluten detection kit using green fluroscent protein in PQLP aminoacid sequence. If kumamolisin cuts the gluten it will have cut GFP as well so there will be no colour but if it does not cut the gluten than it also has not cut GFP so green color will be observed.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 2646
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1397
Illegal NgoMIV site found at 1670
Illegal NgoMIV site found at 2198
Illegal NgoMIV site found at 2444
Illegal AgeI site found at 793
Illegal AgeI site found at 905
Illegal AgeI site found at 1722
Illegal AgeI site found at 2298
Illegal AgeI site found at 2538
1000
COMPATIBLE WITH RFC[1000]