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Metabolic engineering of recombinant protein secretion by Saccharomyces cerevisiae.
In this review, the protein post-translational modification such as folding, trafficking, and secretion, steps that are traditionally studied in isolation will here be described in the context of the whole system of protein secretion.
Imbalance of heterologous protein folding and disulfide bond formation rates yields runaway oxidative stress
Apparent runaway oxidative stress due to radical production observed here and elsewhere can be explained by a futile cycle of disulfide formation and breaking that consumes reduced glutathione and produces reactive oxygen species.
Microfluidic screening and whole-genome sequencing identifies mutations associated with improved protein secretion by yeast
By combining high-throughput microfluidic screening with whole-genome sequencing of the selected clones from yeast libraries, the mutations associated with significantly improved protein production are identified and can be used as reverse metabolic engineering target genes in design of efficient cell factories for protein secretion.
Different expression systems for production of recombinant proteins in Saccharomyces cerevisiae
For the IP there is more than 10‐fold higher protein production with the CPOTud vector compared with the standard URA3‐based vector, and this vector system therefore represent a valuable resource for future studies and optimization of recombinant protein production in yeast.
A gRNA-tRNA array for CRISPR-Cas9 based rapid multiplexed genome editing in Saccharomyces cerevisiae
A gRNA-tRNA array for CRISPR-Cas9 (GTR-CRISPR) for multiplexed engineering of Saccharomyces cerevisiae is reported, resulting in a 30-fold increase in free fatty acid production in 10 days using just two-round deletions of eight previously identified genes.
Heat shock response improves heterologous protein secretion in Saccharomyces cerevisiae
The results provide an effective strategy to improve protein secretion and demonstrated an approach that can induce ER and cytosolic chaperones simultaneously.
Metabolic engineering of a synergistic pathway for n-butanol production in Saccharomyces cerevisiae
The engineered yeast strain represents a promising alternative platform for n-butanol production through a synergistic pathway: the endogenous threonine pathway and the introduced citramalate pathway.
High-throughput screening for industrial enzyme production hosts by droplet microfluidics.
A high-throughput method for single cell screening by microfluidic droplet sorting is applied to a whole-genome mutated yeast cell library yielding improved production hosts of secreted industrial
Recent advances in combinatorial biosynthesis for drug discovery
Three major strategies for combinatorial biosynthesis are discussed: 1) precursor-directed biosynthesis; 2) enzyme-level modification, which includes swapping of the entire domains, modules and subunits, site-specific mutagenesis, and directed evolution; 3) pathway-level recombination.