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.
Highlights
In order to expand the capabilities of engineered organisms, synthetic biologists have begun exploring non-model organisms that display characteristics not found in model organisms such as Escherichia coli [1,2,3]
V. natriegens has a growth rate in glucose minimal medium (1.7 h−1 ) that is higher than even E. coli that has undergone adaptive laboratory evolution (~1.0 h−1 ), and 1.0 h−1 appears to be the upper limit for E. coli growth rate [9]
Cells were grown in proprietary minimal media in a microplate with individual wells supplemented by different carbohydrate sources, and cell respiration was determined kinetically by measuring NADH production through reduction of a tetrazolium dye over time
Summary
In order to expand the capabilities of engineered organisms, synthetic biologists have begun exploring non-model organisms that display characteristics not found in model organisms such as Escherichia coli [1,2,3]. The type strain of Vibrio natriegens is a Gram-negative, facultatively anaerobic, salt marsh isolate [4], that has been put forward as an emergent synthetic biology chassis due in large part to its non-pathogenic nature and extremely fast growth rate (reported doubling time < 10 min) [5,6,7,8,9,10,11,12] To put this in context, its growth rate using glucose is at least two times faster than some industrially-relevant microorganisms including E. coli, Bacillus subtilis, Corynebacterium glutamicum, and yeast [7].
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