Use of physiological information and process optimisation enhances production of extracellular nuclease by a marine strain of Bacillus licheniformis

• Published in: Bioresource technology 2013-02, Vol.130, p.552-558
• Main Authors: Rajarajan, Nithyalakshmy, Ward, Alan C., Burgess, J. Grant, Glassey, Jarka
• Format: Article
• Language: English
• Subjects: Bacillus; Bacillus - physiology; Bacillus licheniformis; Bacterial Proteins - biosynthesis; Bacterial Proteins - secretion; biofilm; Biofilms; Biofilms - growth & development; Biological and medical sciences; Biotechnology; Cell Culture Techniques; Construction; Culture Media; Deoxyribonucleases - biosynthesis; Deoxyribonucleases - secretion; Design of experiments; DNA; Enzyme engineering; Enzyme production; enzymes; experimental design; Fundamental and applied biological sciences. Psychology; manganese; Manganese - metabolism; Methods. Procedures. Technologies; Nuclease; Optimisation; Optimization; Phosphates; Phosphates - metabolism; Production of selected enzymes; sporulation; Strain
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2012.12.064

► Production of an antibiofilm nuclease from Bacillus licheniformis was achieved. ► Use of manganese as a secretory stimulus increased NucB production 5-fold. ► Statistical optimisation gave a 10-fold improvement in NucB secretion. ► NucB production was enhanced using a unique physiology driven optimisation approach. The extracellular nuclease, NucB, from Bacillus licheniformis, can digest extracellular DNA in biofilms, causing biofilm dispersal, and may therefore be used commercially to remove biofilms. However, producing quantities of this secreted peptide is difficult and our aim was therefore to improve its laboratory scale production. This study builds on our understanding of B. licheniformis physiology to enhance NucB production. The addition of manganese, which triggers sporulation and enhances NucB expression, lead to a 5-fold increase in NucB production. Optimisation via Placket–Burman design of experiments identified 3 significant medium components and a subsequent Central Composite Design, to determine the optimum levels of these components, resulted in a 10-fold increase to 471U/ml. The optimal phosphate concentration was less than 0.3mM as this is known to inhibit nuclease production. The use of physiologically relevant information combined with optimisation represents a promising approach to increased enzyme production, which may also be widely applicable.