Heterologous production of non-ribosomal peptide LLD-ACV in Saccharomyces cerevisiae
Artikel i vetenskaplig tidskrift, 2009

Non-ribosomal peptides (NRPs) are a diverse family of secondary metabolites with a broad range of biological activities. We started to develop an eukaryotic microbial platform based on the yeast Saccharomyces cerevisiae for heterologous production of NRPs using delta-(L-alpha-aminoadipyl)-L-cysteinyl-D- valine (ACV) as a model NRP. The Penicillium chrysogenum gene pcbAB encoding ACV synthetase was expressed in S. cerevisiae from a high-copy plasmid together with phosphopantetheinyl transferase (PPTase) encoding genes from Aspergillus nidulans, P. chrysogenum and Bacillus subtilis, and in all the three cases production of ACV was observed. To improve ACV synthesis, several factors were investigated. Codon optimization of the 50 end of pcbAB did not significantly increase ACV production. However, a 30-fold enhancement was achieved by lowering the cultivation temperature from 30 to 20 degrees C. When ACVS and PPT a seen coding genes were integrated into the yeast genome, a 6-fold decrease in ACV production was observed indicating that gene copy number was one of the rate-limiting factors for ACV production in yeast. (C) 2009 Elsevier Inc. All rights reserved.

bacillus-subtilis

escherichia-coli

expression

Non-ribosomal peptide synthetase

natural-product

penicillin biosynthesis

Secondary metabolites

gene-cluster

Phosphopantetheinyl transferase

protein

synthetase

S. cerevisiae

low-temperature increases

secondary metabolites

Författare

X. Chen

L. Huang

J. Zhang

Jens B Nielsen

Chalmers, Kemi- och bioteknik, Livsvetenskaper, Systembiologi

Metabolic Engineering

1096-7176 (ISSN) 1096-7184 (eISSN)

Vol. 11 391-397

Ämneskategorier

Industriell bioteknik

Styrkeområden

Livsvetenskaper och teknik

DOI

10.1016/j.ymben.2009.08.002