Engineering the N ‐terminal end of CelA results in improved performance and growth of Caldicellulosiruptor bescii on crystalline cellulose

CelA is the most abundant enzyme secreted by Caldicellulosiruptor bescii and has been shown to outperform mixtures of commercially available exo‐ and endoglucanases in vitro. CelA contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose‐binding domains via linker peptides. Here, repeated aspartate residues were introduced into the N ‐terminal ends of CelA GH9 and GH48 domains to improve secretion efficiency and/or catalytic efficiency of CelA. Among several constructs, the highest activity on carboxymethylcellulose (CMC), 0.81 ± 0.03 mg/mL was observed for the C. bescii strain containing CelA with 5‐aspartate tag at the N ‐terminal end of GH9 domain—an 82% increase over wild type CelA. In addition, expression of CelA with N ‐terminal repeated aspartate residues in C. bescii results in a dramatic increase in its ability to grow on Avicel. Biotechnol. Bioeng. 2017;114: 945–950. © 2016 Wiley Periodicals, Inc.