Designer Micelles Accelerate Flux Through Engineered Metabolism in E. coli and Support Biocompatible Chemistry

Abstract

Synthetic biology has enabled the production of many value-added chemicals via microbial fermentation. However,the problem of lowproduct titers from recombinant pathwaysh as limited the utility of this approach.M ethods to increase metabolic flux are therefore critical to the success of metabolic engineering.H ere we demonstrate that vitamin Ederived designer micelles,o riginally developed for use in synthetic chemistry,a re biocompatible and accelerate flux through astyrene production pathway in Escherichia coli. We show that these micelles associate non-covalently with the bacterial outer-membrane and that this interaction increases membrane permeability.I na ddition, these micelles also accommodate both heterogeneous and organic-soluble transition metal catalysts and accelerate biocompatible cyclopropanation in vivo.O verall, this work demonstrates that these surfactants hold great promise for further application in the field of synthetic biotechnology,and for expanding the types of molecules that can be readily accessed from renewable resources via the combination of microbial fermentation and biocompatible chemistry.

Biocatalytic activity of Monascus mycelia depending on physiology and high sensitivity to product concentration

“…Accumulation of high concentration of extracellular Monascus pigments was realized due to the plant oil phase with high extractive capacity. Furthermore, instable/toxic extracellular product can also be converted into relatively stable/nontoxic compound by enzymatic reaction (Willrodt et al 2015 ) or non-enzymatic reaction (Xiong et al 2015 ; Domaille et al 2016 ; Wallace and Balskus 2016 ), which may be a more efficient strategy to make whole cell biocatalyst play its potential.…”

“…In-situ product removal, such as addition of solid-state adsorbent (Evanst and Wang 1984 ), nonaqueous two-phase extraction (Wang and Dai 2010 ), is the most common strategy for elimination of product inhibition/degradation. Cascade conversion of instable/toxic product into stable/non-toxic compound by enzymatic reaction (Willrodt et al 2015 ) or non-enzymatic reaction (Xiong et al 2015 ; Domaille et al 2016 ; Wallace and Balskus 2016 ) is also developed for bioprocess optimization.…”

Interfacing non-enzymatic catalysis with living microorganisms

“…Additionally, modification of metabolites can be an effective method of driving flux through a metabolic pathway towards the product of interest. 51 Furthermore, microbes are able to alter the chemistry of their external environment ( e.g. pH and/or O 2 levels 52 ), potentially facilitating non-enzymatic reactions.…”

Designer metalloenzymes for synthetic biology: Enzyme hybrids for catalysis

“…The extracellular location of transition metal catalysts has been successfully combined with biosynthetic pathways to catalyse the final step in several biosynthetic cascades (Figure 4). This approach has been used to make feedstock chemicals (Figure 4B) [52], small-molecule intermediates (Figure 4C) [53] as well as potential antibiotics (Figure 4D) [54]. Interestingly, as far as the author is aware no engineered metalloenzyme or ArM catalysing an abiotic reaction have been included in a biosynthetic pathway, even though their use in vivo has been described for both ring-closing metathesis and cyclopropanation, as featured in Figure 4B and C.…”