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Production of solubleproteins is essential for structure/functionstudies; however, this usually requires milligram amounts of protein,which can be difficult to obtain with traditional expression systems.Recently, the Gram-negative bacterium Vibrio natriegens emerged as a novel and alternative host platform for productionof proteins in high yields. Here, we used a commercial strain derivedfrom V. natriegens （Vmax X2） to produce soluble bacterialand fungal proteins in milligram scale, which we struggled to achievein Escherichia coli. These proteins include the choleratoxin （CT） and N-acetyl glucosamine-binding proteinA （GbpA） from Vibrio cholerae, the heat-labile enterotoxin（LT） from E. coli and the fungal nematotoxin CCTX2from Coprinopsis cinerea. CT, GbpA, and LT are secretedby the Type II secretion system in their natural hosts. When thesethree proteins were produced in Vmax, they were also secreted andcould be recovered from the growth media. This simplified the downstreampurification procedure and resulted in considerably higher proteinyields compared to production in E. coli （6- to 26-foldincrease）. We also tested Vmax for protein perdeuteration using deuteratedminimal media with deuterium oxide as solvent and achieved a 3-foldincrease in yield compared to the equivalent protocol in E.coli. This is good news, since isotopic labeling is expensiveand often ineffective but represents a necessary prerequisite forsome structural biology techniques. Thus, Vmax represents a promisinghost for production of challenging expression targets and for proteinperdeuteration in amounts suitable for structural biology studies.
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