목차

Chapter 1. General Introduction 1
1.1 Introduction 2
1.2 Aims of This study 6
Chapter 2. Genome resequencing of the Escherichia coli wild-type and mutant XL1-Blue strain 7
2.1 Abstract 8
2.2 Introduction 9
2.3 Materials and Methods 11
2.3.1 Strains and media 11
2.3.2 Culture conditions in flask 11
2.3.3 Carotenoids extraction and quantification 12
2.3.4 Microscopic examination 12
2.3.5 Metabolite analysis 13
2.3.6 Batch fermentation 13
2.3.7 Genome resequencing and Annotation 13
2.4 Results 24
2.4.1 Production of β-carotene in XL1-Blue and Ajou 45 24
2.4.2 Whole genome analysis of wild-type XL1-Blue and Ajou 45 26
2.4.3 Phylogenetic analysis 30
2.4.4 List of resequencing mutation in Ajou 45 32
2.4.5 The knockout of cyaA gene increases β-carotene production 35
2.4.6 A decrease in cAMP increases β-carotene production 40
2.5 Discussion 42
Chapter 3. Transcriptomic analysis of Escherichia coli wild-type and mutant XL1-Blue strains 47
3.1 Abstract 48
3.2 Introduction 50
3.3 Material and Method 52
3.3.1 Strains and media 52
3.3.2 Construction of plasmid 52
3.3.3 Batch fermentation 53
3.3.4 RNA isolation, cDNA library construction, and RNA-seq data analysis 53
3.3.5 Culture conditions in flask 54
3.3.6 Quantification of mRNA by qRT-PCR 54
3.4 Results 60
3.4.1 Batch fermentation of wild type XL1-Blue and mutant XL1-Blue at minimal medium for transcriptomics analysis 60
3.4.2 Read counts of gene regulated by cAMP-CRP complex 62
3.4.3 Overexpression of aroA gene, fucO gene, yqeF gene in complex medium and semi-define medium increases β-carotene production 73
3.4.4 The overexpression of aroA gene, fucO gene and yqeF gene in Escherichia coli, which produces β-carotene with an integration system, increases β-carotene production 78
3.5 Discussion 81
4 Conclusions 83
5 References 85