Titrating bacterial growth and chemical biosynthesis for efficient N-acetylglucosamine and N-acetylneuraminic acid bioproduction

Rongzhen Tian, Yanfeng Liu (), Yanting Cao, Zhongjie Zhang, Jianghua Li, Long Liu, Guocheng Du and Jian Chen ()
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Rongzhen Tian: Jiangnan University
Yanfeng Liu: Jiangnan University
Yanting Cao: Jiangnan University
Zhongjie Zhang: Jiangnan University
Jianghua Li: Jiangnan University
Long Liu: Jiangnan University
Guocheng Du: Jiangnan University
Jian Chen: Jiangnan University

Nature Communications, 2020, vol. 11, issue 1, 1-13

Abstract: Abstract Metabolic engineering facilitates chemical biosynthesis by rewiring cellular resources to produce target compounds. However, an imbalance between cell growth and bioproduction often reduces production efficiency. Genetic code expansion (GCE)-based orthogonal translation systems incorporating non-canonical amino acids (ncAAs) into proteins by reassigning non-canonical codons to ncAAs qualify for balancing cellular metabolism. Here, GCE-based cell growth and biosynthesis balance engineering (GCE-CGBBE) is developed, which is based on titrating expression of cell growth and metabolic flux determinant genes by constructing ncAA-dependent expression patterns. We demonstrate GCE-CGBBE in genome-recoded Escherichia coli Δ321AM by precisely balancing glycolysis and N-acetylglucosamine production, resulting in a 4.54-fold increase in titer. GCE-CGBBE is further expanded to non-genome-recoded Bacillus subtilis to balance growth and N-acetylneuraminic acid bioproduction by titrating essential gene expression, yielding a 2.34-fold increase in titer. Moreover, the development of ncAA-dependent essential gene expression regulation shows efficient biocontainment of engineered B. subtilis to avoid unintended proliferation in nature.

Date: 2020
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DOI: 10.1038/s41467-020-18960-1

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