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

Summary 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. In this study, we describe the successful secretion of a heterologous FAD cofactor‐containing enzyme into the supernatant of Corynebacterium glutamicum by using the twin‐arginine translocation (Tat) protein export machinery for protein translocation across the cytoplasmic membrane. Our results show for the first time that, also for cofactor‐containing proteins, a secretory production strategy is a feasible and promising option.