Abstract: Disclosed herein are systems, methods and compositions of matter for engineered formate dehydrogenases electrocatalytically driven in reverse to fix CO2 to formate, an electrode interface composed of self-assembled monolayers (SAM), and a nanowire link between the electrode and the enzyme's iron sulfur (FeS) cluster (engineered) to specifically link the iron sulfur cluster to the electrode. Engineering the interface between the enzyme and the electrode enables a high degree of activity and function compared to just an enzyme immobilized on the electrode. This interface enables specific orientational control of the enzyme and allows for separation of the enzyme from the electrode surface to allow retention of its native hydrodynamic radius. The transfer of electrons via the nanowire to the FeS cluster allows the enzyme to utilize its native electron transfer pathways.

Abstract: A range of Cel7 putative cellobiohydrolase genes were identified using genome mining and homologous sequence alignment to Cel7A from Trichoderma reesei. A representative subset of these genes from across a broad diversity of evolutionarily disparate sources were cloned and expressed in T. reesei using a constitutive promotor and a common secretion signal. The purified recombinant enzymes were tested for efficacy on various substrates. The top performers were subjected to structural studies and subsites likely to confer enhanced performance were predicted using homology modeling and comparisons of natural sequence diversity. Once identified, the subsites were genetically introduced individually and combinatorically into the best in class Cel7A backbone we have found to date and then expressed in T. reesei and tested. A triple mutant was determined to have the highest cellulase activity we have measured in a cellobiohydrolase to date.

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: Disclosed herein are methods of making multifunctional microbial cellulases. The engineered multifunctional microbial cellulases disclosed herein exhibit improved activity over native cellulases.

Abstract: Disclosed herein are methods of making multifunctional microbial cellulases. The engineered multifunctional microbial cellulases disclosed herein exhibit improved activity over native cellulases.

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: Disclosed herein are enzymes and combinations of the enzymes useful for the hydrolysis of cellulose and the conversion of biomass. Methods of degrading cellulose and biomass using enzymes and cocktails of enzymes are also disclosed.

Abstract: Disclosed herein are enzymes and combinations of the enzymes useful for the hydrolysis of cellulose and the conversion of biomass. Methods of degrading cellulose and biomass using enzymes and cocktails of enzymes are also disclosed.

Abstract: Improved systems and methods for reducing costs and increasing yields of cellulosic ethanol are disclosed herein, along with plants genetically transformed for increased biomass, expression of lignocellulolytic enzyme polypeptides, and/or simplification of harvesting and downstream processing. Methods for processing biomass from these transgenic plants that involve less severe and/or less expensive pre-treatment protocols than are typically employed are also disclosed.

Abstract: The disclosure provides reactors for rapid pretreatment of multiple biomass samples in a simple, process-driven, high throughput screening assay. This disclosure also provides methods and systems for rapid, high-throughput pretreatment and subsequent enzyme hydrolysis testing of multiple biomass samples.