Currently available tools for multiplex bacterial genome engineering are optimized for a few laboratory model strains, demand extensive prior modification of the host strain, and lead to the accumulation of numerous off-target modifications. Building on prior development of multiplex automated genome engineering (MAGE), our work addresses these problems in a single framework. Using a dominant-negative mutant protein of the methyl-directed mismatch repair (MMR) system, we achieved a transient suppression of DNA repair in Escherichia coli, which is necessary for efficient oligonucleotide integration. By integrating all necessary components into a broad-host vector, we developed a new workflow we term pORTMAGE. It allows efficient modification of multiple loci, without any observable off-target mutagenesis and prior modification of the host genome. Because of the conserved nature of the bacterial MMR system, pORTMAGE simultaneously allows genome editing and mutant library generation in other biotechnologically and clinically relevant bacterial species. Finally, we applied pORTMAGE to study a set of antibiotic resistance-conferring mutations in Salmonella enterica and E. coli. Despite over 100 million y of divergence between the two species, mutational effects remained generally conserved. In sum, a single transformation of a pORTMAGE plasmid allows bacterial species of interest to become an efficient host for genome engineering. These advances pave the way toward biotechnological and therapeutic applications. Finally, pORTMAGE allows systematic comparison of mutational effects and epistasis across a wide range of bacterial species.

https://www.pnas.org/content/pnas/113/9/2502.full.pdf

A highly precise and portable genome engineering method allows comparison of mutational effects across bacterial species.

Bioconversion technologies of crude glycerol to value added industrial products

Metabolic engineering has been enabling development of high performance microbial strains for the efficient production of natural and non-natural compounds from renewable non-food biomass. Even though microbial production of various chemicals has successfully been conducted and commercialized, there are still numerous chemicals and materials that await their efficient bio-based production. Aromatic chemicals, which are typically derived from benzene, toluene and xylene in petroleum industry, have been used in large amounts in various industries. Over the last three decades, many metabolically engineered microorganisms have been developed for the bio-based production of aromatic chemicals, many of which are derived from aromatic amino acid pathways. This review highlights the latest metabolic engineering strategies and tools applied to the biosynthesis of aromatic chemicals, many derived from shikimate and aromatic amino acids, including l-phenylalanine, l-tyrosine and l-tryptophan. It is expected that more and more engineered microorganisms capable of efficiently producing aromatic chemicals will be developed toward their industrial-scale production from renewable biomass.

/pdf/metabolic-engineering-of-microorganisms-for-production-of-1g2zoct0qj.pdf

Metabolic engineering of microorganisms for production of aromatic compounds

Advances in Chromobacterium violaceum and properties of violacein-Its main secondary metabolite: A review

Building microbial factories for the production of aromatic amino acid pathway derivatives: From commodity chemicals to plant-sourced natural products

Fed-batch fermentation of recombinant Citrobacter freundii with expression of a violacein-synthesizing gene cluster for efficient violacein production from glycerol

Heterologous expression of particulate methane monooxygenase in different host cells

The invention discloses a method for preparing violacein and a special bacterium thereof. A recombinant methylobacterium extorquens is obtained by introducing a violacein biosynthesis-related gene cluster into a host of methylobacterium extorquens. The method for preparing violacein of the invention is low in cost; on one hand, a medium used for fermentation is simple in component and low in cost; on the other hand, the method of the invention only adopts methanol as a carbon source to obtain violacein by fermentation, and methanol can be synthesized by petroleum or biological renewable resources, and is low in price; the method of the invention realizes industrial production of violacein by using cheap and renewable resources; the method of the invention is simple in operation, high in yield, and can reach 0.15 g/l fermentation liquor. The recombinant bacterium of the invention has good effect in producing violacein. Therefore, the recombinant bacterium and the preparation method of violacein of the invention have wide application prospects.

Method for preparing violacein and special bacterium thereof

The invention discloses a method for preparing a pigment In the method, sodium dodecyl sulfate serving as a surfactant is added into fermentation liquid in the fermentation process of a genetic engineering strain Cfreundii/pComvio; and the pigment consists of violacein and deoxidized violacein In the method, after the sodium dodecyl sulfate serving as the surfactant is added into the fermentation liquid, a reaction substrate is added into the fermentation liquid After the surfactant is added in the fermentation process of the genetic engineering strain Cfreundii/pComvio, the strain growsrapidly and is easy to culture, the biosynthesis stability of the pigment is high, the yield is improved and the fermentation period is shortened The pigment yield of thalli can reach 21+/-01896gL fermentation liquid under the optimal treatment conditions

Method for preparing pigment

Disruption of lactate dehydrogenase and alcohol dehydrogenase for increased hydrogen production and its effect on metabolic flux in Enterobacter aerogenes

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