A passive, hydrodynamic technique implemented using a microfluidic device to perform co-encapsulation of samples in droplets and sorting of said droplets is described herein. The hydrodynamic technique utilizes laminar flows and high shear liquid-liquid interfaces at a microfluidic junction to encapsulate samples in the droplets. A sorting mechanism is implemented to separate sample droplets from empty droplets. This technique can achieve a one-one-one encapsulation efficiency of about 80% and can significantly improve the droplet sequencing and related applications in single cell genomics and proteomics.

A passive, hydrodynamic technique implemented using a microfluidic device to perform co-encapsulation of samples in droplets and sorting of said droplets is described herein. The hydrodynamic technique utilizes laminar flows and high shear liquid-liquid interfaces at a microfluidic junction to encapsulate samples in the droplets. A sorting mechanism is implemented to separate sample droplets from empty droplets. This technique can achieve a one-one-one encapsulation efficiency of about 80% and can significantly improve the droplet sequencing and related applications in single cell genomics and proteomics.

A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

A method of proteolyzing a protein, including immobilizing a protein in at least one pore of a porous body, and contacting the protein immobilized in the pore and a protease immobilized on a solid surface such that the protease selectively accesses a site of the protein and proteolyzes the protein at the site.

A method of proteolyzing a protein, including immobilizing a protein in at least one pore of a porous body, and contacting the protein immobilized in the pore and a protease immobilized on a solid surface such that the protease selectively accesses a site of the protein and proteolyzes the protein at the site.

Polypeptide scaffolds comprising enzymatic proteins are provided. The enzymatic polypeptide scaffolds comprise heterologous enzymes to form a heterologous metabolic pathway, and can be targeted to a substrate through a surface anchoring domain. The enzymatic polypeptide scaffolds leverage the high specificity and affinity protein/protein interaction between the cohesins and dockerins of microorganismal cellulosomes to form custom enzymatic arrays.

Polypeptide scaffolds comprising enzymatic proteins are provided. The enzymatic polypeptide scaffolds comprise heterologous enzymes to form a heterologous metabolic pathway, and can be targeted to a substrate through a surface anchoring domain. The enzymatic polypeptide scaffolds leverage the high specificity and affinity protein/protein interaction between the cohesins and dockerins of microorganismal cellulosomes to form custom enzymatic arrays.

A method of proteolyzing a protein, including immobilizing a protein in at least one pore of a porous body, and contacting the protein immobilized in the pore and a protease immobilized on a solid surface such that the protease selectively accesses a site of the protein and proteolyzes the protein at the site.

A method of proteolyzing a protein, including immobilizing a protein in at least one pore of a porous body, and contacting the protein immobilized in the pore and a protease immobilized on a solid surface such that the protease selectively accesses a site of the protein and proteolyzes the protein at the site.

Provided are amino acid sequences of engineered transaminase polynucleotide that are useful for asymmetrically synthesizing chiral amine compounds, and its preparation process and reaction process under industrial-relevant conditions. Also provided are polynucleotide sequences encoding engineered transaminase polypeptides, engineered host cells capable of expressing engineered transaminase polypeptides, and methods of producing chiral amine compounds using engineered transaminase polypeptides. Compared to other enzymes, the engineered transaminase polypeptides provided by this invention have better catalytic activity and stability, and are not inhibited by chiral amine products in the synthesis process. The use of the engineered polypeptides of the present invention for the preparation of chiral amine compound results in higher unit activity, and has good industrial application prospects.