Efficient strategies to enhance plasmid stability for fermentation of recombinant Escherichia coli harboring tyrosine phenol lyase View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2021-04-08

AUTHORS

Xiao-Ling Tang, Wen-Ye Hu, Zhi-Chao Wang, Ren-Chao Zheng, Yu-Guo Zheng

ABSTRACT

ObjectiveTo solve the bottleneck of plasmid instability during microbial fermentation of l-DOPA with recombinant Escherichia coli expressing heterologous tyrosine phenol lyase.ResultsThe 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.ConclusionsThe 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. More... »

PAGES

1265-1276

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s10529-021-03082-5

DOI

http://dx.doi.org/10.1007/s10529-021-03082-5

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1137000367

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/33830386


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