Artificial assembly of a minimal cell.
ABSTRACT Synthetic Biology approaches can assemble and/or reconstruct cell parts in synthetic compartments. A minimal cell as a model for early living cells can be artificially constructed in the laboratory resuming the main properties of a basic cell living system: a synthetic cell compartment or liposome to host a minimal metabolism based on protein synthesis, and a shell and core reproduction mechanism, all in an artificial cell assembly and remaining in the realm of minimal living. It is becoming realistic to construct artificial cells, starting from a minimal cell assembly, and deliver cell-like bioreactors to synthesize pure proteins/enzymes or isolate single pathways. These artificial cell-like systems could perform different tasks in antimicrobial drug development, drug delivery and diagnostic applications.
Article: Multiplexed in vivo His-tagging of enzyme pathways for in vitro single-pot multi-enzyme catalysis.[show abstract] [hide abstract]
ABSTRACT: Protein pathways are dynamic and highly coordinated spatially and temporally, capable of performing a diverse range of complex chemistries and enzymatic reactions with precision and at high efficiency. Biotechnology aims to harvest these natural systems to construct more advanced in vitro reactions, capable of new chemistries and operating at high yield. Here, we present an efficient Multiplex Automated Genome Engineering (MAGE) strategy to simultaneously modify and co-purify large protein complexes and pathways from the model organism Escherichia coli to reconstitute functional synthetic proteomes in vitro. By application of over 110 MAGE cycles, we successfully inserted hexa-histidine sequences into 38 essential genes in vivo that encode for the entire translation machinery. Streamlined co-purification and reconstitution of the translation protein complex enabled protein synthesis in vitro. Our approach can be applied to a growing area of applications in in vitro one-pot multi-enzyme catalysis (MEC) to manipulate or enhance in vitro pathways such as natural product or carbohydrate biosynthesis.ACS synthetic biology. 02/2012; 1(2):43-52.
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ABSTRACT: Minimal cells comprise only the genes and biomolecular machinery necessary for basic life. Synthesizing minimal and minimized cells will improve understanding of core biology, enhance development of biotechnology strains of bacteria, and enable evolutionary optimization of natural and unnatural biopolymers. Design and construction of minimal cells is proceeding in two different directions: 'top-down' reduction of bacterial genomes in vivo and 'bottom-up' integration of DNA/RNA/protein/membrane syntheses in vitro. Major progress in the past 5 years has occurred in synthetic genomics, minimization of the Escherichia coli genome, sequencing of minimal bacterial endosymbionts, identification of essential genes, and integration of biochemical systems.Current opinion in biotechnology 10/2010; 21(5):697-703. · 7.82 Impact Factor