Scale-up of industrial microbial processes.

Scaling up industrial microbial processes for commercial production is a high-stakes endeavor, requiring time and investment often exceeding that for laboratory microbe and process development. Omissions, oversights and errors can be costly, even fatal to the program. Approached properly, scale-up can be executed successfully. Three guiding principles are provided as a basis: begin with the end in mind; be diligent in the details; prepare for the unexpected. A detailed roadmap builds on these principles. There is a special emphasis on the fermentation step, which is usually the costliest and also impacts downstream processing. Examples of common scale-up mistakes and the recommended approaches are given. It is advised that engineering resources skilled in integrated process development and scale-up be engaged from the very beginning of microbe and process development to guide ongoing R&D, thus ensuring a smooth and profitable path to the large-scale commercial end.

An integrated biotechnology platform for developing sustainable chemical processes.

Genomatica has established an integrated computational/experimental metabolic engineering platform to design, create, and optimize novel high performance organisms and bioprocesses. Here we present our platform and its use to develop E. coli strains for production of the industrial chemical 1,4-butanediol (BDO) from sugars. A series of examples are given to demonstrate how a rational approach to strain engineering, including carefully designed diagnostic experiments, provided critical insights about pathway bottlenecks, byproducts, expression balancing, and commercial robustness, leading to a superior BDO production strain and process.

Barrier properties of gastrointestinal mucus to nanoparticle transport.

Gastrointestinal mucus, a complex network of highly branched glycoproteins and macromolecules, is the first barrier through which orally delivered drug and gene vectors must traverse. The diffusion of such vectors can be restricted by the high adhesivity and viscoelasticity of mucus. In this investigation, the barrier properties of gastrointestinal mucus to particle transport were explored using real-time multiple particle tracking. The influence of surface chemistry on particle transport rates was examined using amine-, carboxylate-, and sulfate-modified polystyrene nanoparticles. A strong dependence of particle mobility in gastrointestinal mucus on surface charge was observed, with anionic particles diffusing 20-30 times faster than cationic particles. Comparison of diffusion coefficients calculated for gastrointestinal mucus with significantly varying values previously reported in the literature for other mucus sources, including cervicovaginal mucus and cystic fibrosis sputum, highlight the dependence of mucus barrier properties on the anatomical source. A significant degree of transport rate heterogeneity was also observed in native gastrointestinal mucus, suggesting a highly heterogeneous distribution of pore sizes. Furthermore, the suitability of purified mucin as a model system for transport studies was assessed by comparing particle transport rates between native intestinal mucus and purified porcine gastric mucin. Particle transport rates were approximately threefold lower in native mucus compared to purified mucin for anionic particles, yet comparable for cationic particles. Differences between barrier properties of the purified mucin preparation and native mucus depended on specific carrier properties, indicating that the purified mucin preparation does not provide an accurate model system for native mucus.