Abstract: Disclosed herein are methods and compositions for a new way to separate biologics using coated pegboards—plastic plates affixed with an array of 96-pegs which align with the wells of a traditional 96-well plate used in bench top experiments. The ends of the pegs are conjugated to various compounds which are able to bind tags on engineered proteins (ex. Ni-NTA would bind a His-Tag on an engineered protein). These pegs extend into plate wells to specifically bind the tagged protein from the cell lysate which contains hundreds of other proteins. The pegboard, now laden with the protein of interest (POI), may then be dipped into sequential buffers to wash away potential non-specifically bound proteins to purify the POI.

Abstract: Disclosed herein is a novel, cofactor balanced, cell-free biocatalysis pathway to make diacids from formaldehyde or methanol that does not use ATP and produces no CO2. In another embodiment, disclosed herein is a novel C5/C6 (hydrolysate) utilization pathway that interplays with the cell making diacids from formaldehyde or methanol that does not use ATP, produces no CO2, and replaces ATP with (cheap) polyphosphate (no other cofactors needed). Using methods and compositions disclosed herein, no CO2 is produced and the pathways disclosed herein are cell-free. The combination of pathways, feedstocks and products disclosed herein is novel.

Abstract: 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.

Abstract: 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.