Overexpression of a designed 2.2 kb gene of eukaryotic phenylalanine ammonia-lyase in Escherichia coli

被引：48

作者：

Baedeker, M ^{[1
]}

Schulz, GE ^{[1
]}

机构：

[1] Univ Freiburg, Inst Organ Chem & Biochem, D-79104 Freiburg, Germany

来源：

FEBS LETTERS | 1999年 / 457卷 / 01期

关键词：

gene synthesis; phenylpropanoid metabolism; HSP-60 chaperone coexpression; codon usage; inclusion body;

D O I：

10.1016/S0014-5793(99)01000-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

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 transcinnamate. 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. (C) 1999 Federation of European Biochemical Societies.

引用

页码：57 / 60

页数：4

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