Exploiting the Feedstock Flexibility of the Emergent Synthetic Biology Chassis Vibrio natriegens for Engineered Natural Product Production

Abstract

A recent goal of synthetic biology has been to identify new chassis that provide benefits lacking in model organisms. Vibrio natriegens is a marine Gram-negative bacterium which is an emergent synthetic biology chassis with inherent benefits: An extremely fast growth rate, genetic tractability, and the ability to grow on a variety of carbon sources ("feedstock flexibility"). Given these inherent benefits, we sought to determine its potential to heterologously produce natural products, and chose beta-carotene and violacein as test cases. For beta-carotene production, we expressed the beta-carotene biosynthetic pathway from the sister marine bacterium Vibrio campbellii, as well as the mevalonate biosynthetic pathway from the Gram-positive bacterium Lactobacillus acidophilus to improve precursor abundance. Violacein was produced by expressing a biosynthetic gene cluster derived from Chromobacterium violaceum. Not only was V. natriegens able to heterologously produce these compounds in rich media, illustrating its promise as a new chassis for small molecule drug production, but it also did so in minimal media using a variety of feedstocks. The ability for V. natriegens to produce natural products with multiple industrially-relevant feedstocks argues for continued investigations into the production of more complex natural products in this chassis.

Metabolic engineering of Vibrio natriegens

“…More than 60 different carbon sources (including various of industrial relevance) can be utilized by V. natriegens as sole source of carbon and energy under aerobic conditions and at least d-gluconate, d-glucose, d-mannitol, and d-ribose support biomass generation in anaerobic cultivations [5,9,16,21]. Further studies suggest moreover, that various substrates do not support growth but are metabolized by V. natriegens as detected by respiratory activity of the cells [10,25] or reduction of alternative terminal electron acceptors like Fe(III) citrate [24]. Table 3 provides an overview of all carbohydrates and polymers, amino and carboxylic acids, alcohols and polyamines, that promote growth of V. natriegens under aerobic and anaerobic conditions or that are metabolized without supporting growth [5,9,10,16,21,24,25].…”

Engineering Vibrio sp. SP1 for the production of carotenoids directly from brown macroalgae

“…4 C). Previous attempt to produce carotenoids from Vibrio natriegens yielded 0.6 mg L −1 of β -carotene from 4 g L −1 glucose [16] . Considering this strain had additional genetic engineering to integrate genes regarding MVA pathway from Lactobacillus acidophilus to replenish the amount of substrate and the substrate was glucose, the overall productivity of β -carotene from alginate in the engineered Vibrio sp.…”

“…Among various Vibrio species, the most commonly studied and engineered strain is Vibrio natriegens [13] . Metabolic engineering for the bioproduction enabled the strain to produce high value-added chemicals such as melanin [14] , selenium nanoparticle [15] , β -carotene and violacein [16] . However, despite the merit of the strain, it lacks ability to assimilate third generation biomass, alginate, a prominent sugar in brown macroalgae.…”

Complete Genome Sequences of Two Vibrio natriegens Bacteriophages

“…With a doubling time of less than 10 minutes under optimal conditions, it is the fastest-growing bacterium ( 2 ). It has garnered interest as a biotechnology platform for protein production, molecular biology, and metabolic engineering ( 3 – 6 ). Identifying bacteriophage that infect V. natriegens has the potential to aid in these efforts, as molecular genetic tools are often derived from bacteriophage ( 7 ).…”