title: Development of a Novel Plasmid-Free Thymidine Producer by Reprogramming Nucleotide Metabolic Pathways.
abstract: A novel thymidine-producing strain of Escherichia coli was prepared by genome recombineering. Eleven genes were deleted by replacement with an expression cassette, and 7 genes were integrated into the genome. The resulting strain, E. coli HLT013, showed a high thymidine yield with a low deoxyuridine content. DNA microarrays were then used to compare the gene expression profiles of HLT013 and its isogenic parent strain. Based on microarray analysis, the pyr biosynthesis genes and 10 additional genes were selected and then expressed in HLT013 to find reasonable candidates for enhancing thymidine yield. Among these, phage shock protein A (PspA) showed positive effects on thymidine production by diminishing redox stress. Thus, we integrated pspA into the HLT013 genome, resulting in E. coli strain HLT026, which produced 13.2 g/liter thymidine for 120 h with fed-batch fermentation. Here, we also provide a basis for new testable hypotheses regarding the enhancement of thymidine productivity and the attainment of a more complete understanding of nucleotide metabolism in bacteria.
authors: ForBioKorea Co., Ltd., Seoul, Republic of Korea.
authors: Jin-Sook
authors: JS
authors: Kim
authors: BioNgene Co., Ltd., Seoul, Republic of Korea.
authors: Min-Kyung
authors: MK
authors: Jeong
authors: ForBioKorea Co., Ltd., Seoul, Republic of Korea.
authors: Bong-Seong
authors: BS
authors: Koo
authors: ForBioKorea Co., Ltd., Seoul, Republic of Korea BioNgene Co., Ltd., Seoul, Republic of Korea hclee0609@gmail.com.
authors: Hyeon-Cheol
authors: HC
authors: Lee
mesh_terms: D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering; D053858:Metabolic Networks and Pathways; D009711:Nucleotides / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D013936:Thymidine / Q000378:metabolism*
chemical_list: D009711:Nucleotides; D013936:Thymidine