Abstract
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.
Highlights
Bacterial cellulose (BC) is devoid of hemicellulose and lignin, which are the common contaminants of plant cellulose, making it the purest forms of cellulose obtainable as-produced and is desirable for greener synthesis option
We validate the applicability of the platform using three different strains that are Gluconacetobacter hansenii ATCC 53582, Gluconacetobacter xylinus ATCC 700178 and Komagataeibacter rhaeticus iGEM, under different growth conditions
Based on the previously discussed Keller-Segel surface growth model of bacteria, we expected that the increase in the bacterial population at the site of inoculation would lead to depletion of nutrients with time
Summary
BC is devoid of hemicellulose and lignin, which are the common contaminants of plant cellulose, making it the purest forms of cellulose obtainable as-produced and is desirable for greener synthesis option. A platform for quick screening of optimal cellulose producing conditions is beneficial in circumventing practical difficulties associated with culturing of the bacteria by providing answers to some of these biological questions: Does the bacteria have preference towards carbon source and environmental conditions for growth or cellulose production? Surface growth of chemotactic bacteria has been previously studied by several groups with Keller-Segel type as the widely accepted model[8,9]. In such experimental setup, a small drop of bacterial inoculum is added on top of a soft-agar medium surface at the centre of a petri-dish/plate and the bacteria is allowed to grow. We further establish that the trends obtained through the surface-growth platform correlate well with cellulose yields achievable under static culture conditions
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