Difference between revisions of "Part:BBa K2455007"

Latest revision as of 18:39, 28 October 2017

YddG-TrpE construct with an E. coli RBS between (BBa_K2455004 & BBa_K2455001)

This is a composite part consisting of the two YddG (BBa_K2455004) and TrpE (BBa_K2455001) biobricks with an RBS in between.

This biobrick has been designed in order to over-express and secrete tryptophan in E. coli.

YddG is an aromatic amino acid exporter that is responsible for the secretion of Tryptophan, Tyrosine and Phenylalanine. Previous work by Wang et al. have shown that over-expression of YddG in E.coli increases the accumulation of Tryptophan in the culture medium.

TrpE codes for the Anthranilate enzyme responsible for the first step of the conversion of Chorismate into to tryptophan. Previous work (Gu et al., 2012) have shown that over-expression of TrpE leads to increased tryptophan production in E. coli. The trpE gene carries a M293T point mutation (to avoid negative feedback inhibition) and a hexa histidine-tag attached by a short linker sequence.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1876
Illegal BglII site found at 1937
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 577
Illegal AgeI site found at 639
Illegal AgeI site found at 756
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2455007 short</partinfo>
-This is a composite part consisting of the two YddG ([[Part:BBa_K2455004|BBa_K2455004]]) and AroG ([[Part:BBa_K2455000|BBa_K2455000]]) biobricks with an RBS in between.
+This is a composite part consisting of the two YddG ([[Part:BBa_K2455004|BBa_K2455004]]) and TrpE ([[Part:BBa_K2455001|BBa_K2455001]]) biobricks with an RBS in between.
-This biobrick has been designed in order to over-express production of tryptophan, tyrosine and phenylalanine in <i>E. coli</i>.
+This biobrick has been designed in order to over-express and secrete tryptophan in <i>E. coli</i>.
 YddG is an aromatic amino acid exporter that is responsible for the secretion of Tryptophan, Tyrosine and Phenylalanine. Previous work by Wang et al. have shown that over-expression of YddG in <i>E.coli</i> increases the accumulation of Tryptophan in the culture medium.
-AroG gene from <i>E. coli</i> strain MG1655 carrying an P150L point mutation (to avoid negative feedback inhibition) and a hexa histidine-tag attached by a short linker sequence. AroG is the first enzyme of the shikimate pathway leading to synthesis of chorismate, which subsequently can be converted to tryptophan, tyrosine, and phenylalanine. Being the first enzyme in the pathway, it determines the carbon flow towards amino acid synthesis and thus the production. Previous work (Gu <i>et al.</i>, 2012) have shown that overexpression of AroG leads to increased tryptophan production in <i>E. coli.</i>
+TrpE codes for the Anthranilate enzyme responsible for the first step of the conversion of Chorismate into to tryptophan. Previous work ([https://microbialcellfactories.biomedcentral.com/articles/10.1186/1475-2859-11-30 Gu <i>et al.</i>, 2012]) have shown that over-expression of TrpE leads to increased tryptophan production in E. coli. The trpE gene carries a M293T point mutation (to avoid negative feedback inhibition) and a hexa histidine-tag attached by a short linker sequence.
 <!-- Add more about the biology of this part here