Metabolic engineering of Vibrio natriegens for the production of N-acetylglucosamine

Abstract

Vibrio natriegens has emerged as a promising microbial chassis for industrial biotechnology due to its unparalleled growth rate and metabolic versatility. In this study, V. natriegens ATCC14048 was rationally engineered as a high-efficiency platform for N-acetylglucosamine (GlcNAc) biosynthesis. First, key biosynthetic genes including glmS from Escherichia coli and Scgna1 from Saccharomyces cerevisiae were overexpressed, while the native GlcNAc degradation gene nagA was knocked out, which led to a GlcNAc titer of 0.11 g/L. Subsequently, blocking the pentose phosphate pathway and glycolysis enhanced precursor availability and achieved GlcNAc titer of 1.22 g/L. Metabolic flux was further amplified by deleting rapZ, nagB, and manX genes, along with chromosomal integration of the strong promoter J23119 upstream of endogenous glmS, yielding 3.59 g/L GlcNAc. Shake flask cultivation optimization through nitrogen source addition and oxygen supply ultimately elevated GlcNAc titer to 6.89 g/L in shake-flask cultures. This work demonstrates V. natriegens' exceptional metabolic plasticity and positions it as a superior chassis for industrial-scale biomanufacturing of amino sugars.