Whole-cell biocatalyst for cadaverine production using stable, constitutive and high expression of lysine decarboxylase in recombinant Escherichia coli W3110

• Published in: Enzyme and microbial technology Vol. 148; p. 109811
• Main Authors: Ting, Wan-Wen, Huang, Chih-Yu, Wu, Pong-Yee, Huang, Shih-Fang, Lin, Hung-Yi, Li, Sheng-Feng, Chang, Jo-Shu, Ng, I-Son
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01-08-2021
• Subjects: Cadaverine; Constitutive expression; E. coli W3110; Lysine decarboxylase; Whole-cell biocatalyst; Cadaverine; Whole-cell biocatalyst; Lysine decarboxylase; E. coli W3110; Constitutive expression
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/j.enzmictec.2021.109811

•J23100 promoter was suitable for lysine decarboxylase expression in E. coli W3110.•Strain JW showed the excellent plasmid stability even in antibiotic free condition.•Maximal activity of biocatalyst was achieved by using fed-batch fermentation.•A 221 g/L titer and 55.3 g/L-h productivity of cadaverine was obtained.•The biocatalyst can be reused effectively with minimal biocatalyst addition.
Microbial production of industrial chemicals is a sustainable approach to reduce the dependence on petroleum-based chemicals such as acids, alcohols, and amines, in which the cadaverine is a natural diamide and serves as one of the key monomers for biopolymer production. In this study, the constitutive promoter J23100 driven lysine decarboxylase (CadA) for cadaverine production was established and compared in different Escherichia coli strains. The best chassis designed as JW, expressed the highest amount of CadA by using J23100 promoter, showing stable and high copy numbers (i.e., PCN > 100) when culture in the antibiotic-free medium. JW attained a CadA activity of 167 g-DAP/g-DCW-h and had the maximum biocatalyst of 45.6 g-DCW/L in fed-batch fermentation. In addition, JW was able to convert 2.5 M L-lysine to 221 g/L cadaverine, with 86 % yield and 55.3 g/L-h productivity. The whole-cell biocatalyst could be reused over four times at an average of 97 % conversion when supplied half of fresh cells in the reaction. This work developed a stable, constitutive expression, long-term preservation, high-level expression of CadA for DAP production, and paved an alternative opportunity of bio-nylon for industry in the future.