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Literature DB >> 19557307

Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression.

Zhong-Liu Wu¹, Jing Qiao, Zhi-Gang Zhang, F Peter Guengerich, Yan Liu, Xiao-Qiong Pei.

Abstract

To elucidate the effects of codon optimization and chaperone coexpression on the heterologous expression of mammalian cytochrome P450s (P450) in Escherichia coli, the expression of P450s 2B1, 2S1, 2U1, 2W1, and 27C1 were investigated. With codon optimization for N-terminus or the entire gene, the expression levels of P450 27C1, 2U1 and 2W1 increased 22-fold, 3.6-fold and 2.1-fold, respectively, while those for P450s 2B1 and 2S1 remained unchanged. With coexpression of E. coli molecular chaperones GroEL/ES, the expression level increased up to 14-fold for P450 27C1, and 3- to 5-fold for P450s 2B1, 2S1, and 2W1. Simultaneous application of these two techniques resulted in synergetic effects.

Entities: Chemical Disease Gene Species

Mesh：

Substances：

Year: 2009 PMID： 19557307 PMCID： PMC4188451 DOI： 10.1007/s10529-009-0059-5

Source DB: PubMed Journal: Biotechnol Lett ISSN： 0141-5492 Impact factor: 2.461

15 in total

1. Hierarchy of sequence-dependent features associated with prokaryotic translation.

Authors: Gila Lithwick; Hanah Margalit
Journal: Genome Res Date: 2003-12 Impact factor: 9.043

2. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE.

Authors: T OMURA; R SATO
Journal: J Biol Chem Date: 1964-07 Impact factor: 5.157

Review 3. Codon bias and heterologous protein expression.

Authors: Claes Gustafsson; Sridhar Govindarajan; Jeremy Minshull
Journal: Trends Biotechnol Date: 2004-07 Impact factor: 19.536

4. A codon window in mRNA downstream of the initiation codon where NGG codons give strongly reduced gene expression in Escherichia coli.

Authors: Ernesto I Gonzalez de Valdivia; Leif A Isaksson
Journal: Nucleic Acids Res Date: 2004-09-30 Impact factor: 16.971

5. Heterologous expression, purification, and properties of human cytochrome P450 27C1.

Authors: Zhong-Liu Wu; Cheryl J Bartleson; Amy-Joan L Ham; F Peter Guengerich
Journal: Arch Biochem Biophys Date: 2005-11-28 Impact factor: 4.013

6. Chaperone coexpression plasmids: differential and synergistic roles of DnaK-DnaJ-GrpE and GroEL-GroES in assisting folding of an allergen of Japanese cedar pollen, Cryj2, in Escherichia coli.

Authors: K Nishihara; M Kanemori; M Kitagawa; H Yanagi; T Yura
Journal: Appl Environ Microbiol Date: 1998-05 Impact factor: 4.792

5. Biosynthetic pathway for the cyanide-free production of phenylacetonitrile in Escherichia coli by utilizing plant cytochrome P450 79A2 and bacterial aldoxime dehydratase.

Authors: Yuta Miki; Yasuhisa Asano
Journal: Appl Environ Microbiol Date: 2014-08-29 Impact factor: 4.792

5 in total

Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression.

1. Hierarchy of sequence-dependent features associated with prokaryotic translation.

2. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE.

Review 3. Codon bias and heterologous protein expression.

4. A codon window in mRNA downstream of the initiation codon where NGG codons give strongly reduced gene expression in Escherichia coli.

5. Heterologous expression, purification, and properties of human cytochrome P450 27C1.

6. Chaperone coexpression plasmids: differential and synergistic roles of DnaK-DnaJ-GrpE and GroEL-GroES in assisting folding of an allergen of Japanese cedar pollen, Cryj2, in Escherichia coli.

7. DNAWorks: an automated method for designing oligonucleotides for PCR-based gene synthesis.

8. Expansion of substrate specificity of cytochrome P450 2A6 by random and site-directed mutagenesis.

9. Coding-sequence determinants of gene expression in Escherichia coli.

10. Expression and enzymatic activity of recombinant cytochrome P450 17 alpha-hydroxylase in Escherichia coli.

1. Oxidation of endogenous N-arachidonoylserotonin by human cytochrome P450 2U1.

2. Cytochrome c adducts with PCB quinoid metabolites.

3. Ligand characterization of CYP4B1 isoforms modified for high-level expression in Escherichia coli and HepG2 cells.

Review 4. Synthetic biology: tools to design, build, and optimize cellular processes.

5. Biosynthetic pathway for the cyanide-free production of phenylacetonitrile in Escherichia coli by utilizing plant cytochrome P450 79A2 and bacterial aldoxime dehydratase.