Difference between revisions of "Part:BBa K2462000"

Revision as of 12:34, 1 November 2017

dehalogenase

Description

   RdhANP is a native protein of Nitratireducror pacificus pht-3B. It is codon-optimized for operated and expression in Bacillus magaterium. RdhANP can catalyze the reduction of ortho-halogenated phenolic compound, which endows it with biological dehalogenation function. Different from other reductive proteins that are membrane-associated and oxygen-sensitive in their family, RdhANP is a soluble cytoplasmic one and oxygen-tolerant, which is fairly good for heterologous expression in other chassis. However, this enzyme displays a strict requirement for o-halogenated phenolic substrates and B12 cofactor; it also should under reductive condition using either reduced methyl viologen or NADPH.

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 678
Illegal BglII site found at 1902
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 90
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 233

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 <partinfo>BBa_K2462000 short</partinfo>
-RdhANP is a native protein of Nitratireducror pacificus pht-3B. It is codon-optimized for operated and expression in Bacillus magaterium. RdhANP can catalyze the reduction of ortho-halogenated phenolic compound, which endows it with biological dehalogenation function. Different from other reductive proteins that are membrane-associated and oxygen-sensitive in their family, RdhANP is a soluble cytoplasmic one and oxygen-tolerant, which is fairly good for heterologous expression in other chassis. However, this enzyme displays a strict requirement for o-halogenated phenolic substrates and B12 cofactor; it also should under reductive condition using either reduced methyl viologen or NADPH.
+===Description===
+    RdhANP is a native protein of Nitratireducror pacificus pht-3B. It is codon-optimized for operated and expression in Bacillus magaterium. RdhANP can catalyze the reduction of ortho-halogenated phenolic compound, which endows it with biological dehalogenation function. Different from other reductive proteins that are membrane-associated and oxygen-sensitive in their family, RdhANP is a soluble cytoplasmic one and oxygen-tolerant, which is fairly good for heterologous expression in other chassis. However, this enzyme displays a strict requirement for o-halogenated phenolic substrates and B12 cofactor; it also should under reductive condition using either reduced methyl viologen or NADPH.
 <!-- Add more about the biology of this part here
-===Usage and Biology===
+===Verification===
+    For the verification of our enzyme, we constructed it into the vector pSPAsp-hp. Then the plasmid containing RdhANP was transformed into Bacillus magaterium through collaboration with FAFU-China. As for the expression, xylose was used as an inducer and the induction lasted for 12 hours under 37℃. After that, 800uM substrate(2,6-DCP was chosen by us as an representative according to the article) was added and interacted with our engineering bacteria for 2 hours under 17℃. Finally, our samples taken before and after the incubation were all tested through HPLC.
+    To be more specific, two groups of bacteria were tested simultaneously with one of them (Bacteria with RdhANP) as experimental group and the other (Bacteria without RdhANP) as a control.
+    The results of HPLC was depicted as following:
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