We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Efficient strategies to enhance plasmid stability for fermentation of recombinant Escherichia coli harboring tyrosine phenol lyase.
- Authors
Tang, Xiao-Ling; Hu, Wen-Ye; Wang, Zhi-Chao; Zheng, Ren-Chao; Zheng, Yu-Guo
- Abstract
Objective: To solve the bottleneck of plasmid instability during microbial fermentation of l-DOPA with recombinant Escherichia coli expressing heterologous tyrosine phenol lyase. Results: The tyrosine phenol lyase from Fusobacterium nucleatum was constitutively expressed in E. coli and a fed-batch fermentation process with temperature down-shift cultivation was performed. Efficient strategies including replacing the original ampicillin resistance gene, as well as inserting cer site that is active for resolving plasmid multimers were applied. As a result, the plasmid stability was increased. The co-use of cer site on plasmid and kanamycin in culture medium resulted in proportion of plasmid containing cells maintained at 100% after fermentation for 35 h. The specific activity of tyrosine phenol lyase reached 1493 U/g dcw, while the volumetric activity increased from 2943 to 14,408 U/L for l-DOPA biosynthesis. Conclusions: The established strategies for plasmid stability is not only promoted the applicability of the recombinant cells for l-DOPA production, but also provides important guidance for industrial fermentation with improved microbial productivity.
- Subjects
TYROSINE; PHENOL; ESCHERICHIA coli; PHENOLS; FERMENTATION; FOSFOMYCIN
- Publication
Biotechnology Letters, 2021, Vol 43, Issue 7, p1265
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-021-03082-5