title: Secretory production of an FAD cofactor-containing cytosolic enzyme (sorbitol-xylitol oxidase from Streptomyces coelicolor) using the twin-arginine translocation (Tat) pathway of Corynebacterium glutamicum.
abstract: Carbohydrate oxidases are biotechnologically interesting enzymes that require a tightly or covalently bound cofactor for activity. Using the industrial workhorse Corynebacterium glutamicum as the expression host, successful secretion of a normally cytosolic FAD cofactor-containing sorbitol-xylitol oxidase from Streptomyces coelicolor was achieved by using the twin-arginine translocation (Tat) protein export machinery for protein translocation across the cytoplasmic membrane. Our results demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies.
authors.affiliation: Institut für Bio- und Geowissenschaften 1, Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Scheele
authors.forename: Dan
authors.initials: D
authors.lastname: Oertel
authors.forename: Johannes
authors.initials: J
authors.lastname: Bongaerts
authors.forename: Stefan
authors.initials: S
authors.lastname: Evers
authors.forename: Hendrik
authors.initials: H
authors.lastname: Hellmuth
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Maurer
authors.forename: Michael
authors.initials: M
authors.lastname: Bott
authors.forename: Roland
authors.initials: R
authors.lastname: Freudl
mesh_terms: D001709:Biotechnology / Q000379:methods*; D048230:Corynebacterium glutamicum / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods; D010088:Oxidoreductases / Q000378:metabolism*; D021381:Protein Transport; D048372:Streptomyces coelicolor / Q000201:enzymology*; D014993:Xylitol / Q000378:metabolism*
chemical_list: D026901:Membrane Transport Proteins; D005182:Flavin-Adenine Dinucleotide; D010088:Oxidoreductases; C111983:sorbitol oxidase; D014993:Xylitol