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4.3.1.7: ethanolamine ammonia-lyase

This is an abbreviated version!
For detailed information about ethanolamine ammonia-lyase, go to the full flat file.

Word Map on EC 4.3.1.7

Reaction

ethanolamine
=
acetaldehyde
+
NH3

Synonyms

AEL, ammonia-lyase, ethanolamine, EAL, ethanolamine ammonia lyase, ethanolamine ammonia-lyase, ethanolamine ammonia-lyase BMC, ethanolamine deaminase, ethanolamine-ammonia lyase, eut-L, EutB, eutBC

ECTree

     4 Lyases
         4.3 Carbon-nitrogen lyases
             4.3.1 Ammonia-lyases
                4.3.1.7 ethanolamine ammonia-lyase

Cloned

Cloned on EC 4.3.1.7 - ethanolamine ammonia-lyase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cloning and overexpression of three different versions (eut-LpET151 his-tagged version (appends a cleavable 33-amino acid tag sequence to the N-terminus), Eut-L_NHIS (short His6-tagged version) and Eut-L_CHIS (short His6-tagged version)) of the protein is carried out directly from the Escherichia coli genome by selective PCR amplification. Selenomethione-derivatized proteins are obtained by growing cloned bacteria in selenomethionine-containing M9 minimal media. Protein overexpression and purification are performed.
-
cloning in Escherichia coli overexpression strain incorporating the cloned Salmonella typhimurium EAL coding sequence
-
Escherichia coli overexpression strain incorporating the cloned Salmonella typhimurium EAL coding sequence
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expression in Escherichia coli
expression in Escherichia coli overexpression strain incorporating the cloned Salmonella typhimurium EAL coding sequences
-
N-terminal His6-tagged beta subunit lacking residues Lysbeta4-Cysbeta43
The plasmid containing the ethanolamine ammonia-lyase coding sequence is extracted from the Escherichia coli overexpression strain. Site-directed mutagenesis is performed on the R160 position within the eutb sequence by using the GeneTailor site-directed mutagenesis kit. Five plasmids, including the four mutations R160A, R160K, R160E, and R160I and a wild-type control, are constructed and transformed into the competent Escherichia coli DH5R T1R strain.
-
the plasmid pET-SEAL, encoding the small and large subunits of EAL is expressed in Escherichia coli
-