title: Functional assembly of a multi-enzyme methanol oxidation cascade on a surface-displayed trifunctional scaffold for enhanced NADH production.
abstract: We report a simple and low-cost strategy that allows the sequential and site-specific assembly of a dehydrogenase-based multi-enzyme cascade for methanol oxidation on the yeast surface using the high-affinity interactions between three orthogonal cohesin-dockerin pairs. The multi-enzyme cascade showed 5 times higher NADH production rate than the non-complexed enzyme mixture, a result of efficient substrate channeling.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE 19716, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.forename: Scott
authors.initials: S
authors.lastname: Banta
authors.forename: Wilfred
authors.initials: W
authors.lastname: Chen
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism; D002480:Cellulase / Q000737:chemistry / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005560:Formate Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000432:Methanol / Q000737:chemistry*; D009243:NAD / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology; D013499:Surface Properties; D000097722:Cohesins
chemical_list: D000426:Alcohol Dehydrogenase; D000445:Aldehyde Oxidoreductases; D018797:Cell Cycle Proteins; D002480:Cellulase; D002868:Chromosomal Proteins, Non-Histone; D004800:Enzymes, Immobilized; D005560:Formate Dehydrogenases; D000432:Methanol; D009243:NAD; D010088:Oxidoreductases; D011994:Recombinant Proteins; D000097722:Cohesins; C087006:formaldehyde dehydrogenase, glutathione-independent