Intelligent optofluidic analysis for ultrafast single bacterium profiling of cellulose production and morphology.

Bacterial cellulose (BC), a renewable type of cellulose, has been used in the manufacture of foods, cosmetics, and biomedical products. To produce BC, a high-throughput single-bacterium measurement is necessary to identify the functional bacteria that can produce BC with sufficient amount and desirable morphology. In this study, a continuous-flow intelligent optofluidic device was developed to enable high-throughput single-bacterium profiling of BC. Single bacteria were incubated in agarose hydrogel particles to produce BC with varied densities and structures. An intelligent convolutional neural network (CNN) computational method was developed to analyze the scattering patterns of BC. The BC production and morphology were determined with a throughput of ∼35 bacteria per second. A total of ∼105 single-bacterium BC samples were characterized within 3 hours. The high flexibility of this approach facilitates high-throughput comprehensive single-cell production analysis for a range of applications in engineering biology.

Rational design of a scalable bioprocess platform for bacterial cellulose production.

Bacterial cellulose (BC) has been gaining importance over the past decades as a versatile material that finds applications in diverse industries. However, a secured supply is hindered by the slow production rate and batch-to-batch variability of the yield. Here, we report a rational approach for characterising the BC production process using Design of Experiment (DoE) methodology to study the impact of different parameters on desired process attributes. Notably, we found that the carbon source used for bacterial growth significantly impacts the interplay between the process variables and affects the desired outcomes. We therefore, propose that the highest priority process outcome in this study, the yield, is a function of the carbon source and optimal reactor design. Our systematic approach has achieved projected BC yields as high as ∼40 g/L for Gluconacetobacter hansenii 53582 grown on sucrose as the carbon source compared to the widely reported yields of ∼10 g/L.

A Novel Platform for Evaluating the Environmental Impacts on Bacterial Cellulose Production.

Bacterial cellulose (BC) is a biocompatible material with versatile applications. However, its large-scale production is challenged by the limited biological knowledge of the bacteria. The advent of synthetic biology has lead the way to the development of BC producing microbes as a novel chassis. Hence, investigation on optimal growth conditions for BC production and understanding of the fundamental biological processes are imperative. In this study, we report a novel analytical platform that can be used for studying the biology and optimizing growth conditions of cellulose producing bacteria. The platform is based on surface growth pattern of the organism and allows us to confirm that cellulose fibrils produced by the bacteria play a pivotal role towards their chemotaxis. The platform efficiently determines the impacts of different growth conditions on cellulose production and is translatable to static culture conditions. The analytical platform provides a means for fundamental biological studies of bacteria chemotaxis as well as systematic approach towards rational design and development of scalable bioprocessing strategies for industrial production of bacterial cellulose.