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Overexpression of a designed 2.2 kb gene of eukaryotic phenylalanine ammonia‐lyase in Escherichia coli
Author(s) -
Baedeker Mathias,
Schulz Georg E
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01000-5
Subject(s) - escherichia coli , phenylalanine ammonia lyase , phenylalanine , biochemistry , gene , enzyme , microbiology and biotechnology , biology , expression vector , chemistry , oligonucleotide , lyase , gene expression , recombinant dna , amino acid
Phenylalanine ammonia‐lyase (EC 4.3.1.5) is a key enzyme in the secondary metabolism of higher plants catalyzing the non‐oxidative conversion of L ‐phenylalanine into trans ‐cinnamate. The nucleotide sequence of its 2.2 kb gene was designed for expression in Escherichia coli and synthesized in a single reaction from 108 oligonucleotides using assembly PCR. After amplification, the gene was cloned into the expression vector pT7‐7 and coexpressed with the chaperone HSP‐60 system. The expression system yielded 70 mg of fully active enzyme per liter culture.