Metabolic Profile of the Genome-Reduced Bacillus subtilis Strain IIG-Bs-27-39: An Attractive Chassis for Recombinant Protein Production

• DOI: 10.1021/acssynbio.4c00254
• Publication Date: 2024-07-09T19:48:04Z
• Authors: Rocío Aguilar Suárez; Michael Kohlstedt; Ayşegül Öktem; Jolanda Neef; Yuzheng Wu; Kaiya Ikeda; Ken-Ichi Yoshida; Josef Altenbuchner; Christoph Wittmann; Jan Maarten van Dijl
• Keywords: Metabolic Engineering; Strain (injury)

The Gram-positive bacterium Bacillus subtilis is extensively used in the industry for secretory production of proteins with commercial value. To further improve its performance, this microbe has been subject extensive genome engineering efforts, especially removal large genomic regions that are dispensable or even counterproductive. Here, we present genome-reduced B. strain IIG-Bs-27-39, which was obtained through systematic deletion mobile genetic elements, as well genes extracellular proteases, sporulation, flagella formation, and antibiotic production. Different from previously characterized strains, IIG-Bs-27-39 still able to grow on minimal media. We feature benchmark against parental 168 respect heterologous protein metabolic parameters during bioreactor cultivation. presented superior secretion difficult-to-produce staphylococcal antigens, higher specific growth rates biomass yields. At level, changes byproduct formation internal amino acid pools were observed, whereas energetic such ATP yield, ATP/ADP levels, adenylate energy charge comparable between two strains. Intriguingly, observed a significant increase total cellular NADPH level all tested conditions increases NAD+ NADP(H) This indicates more available anabolic processes production, thereby providing link physiology performance. On basis, conclude represents an attractive chassis future biotechnological applications.