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ABSTRACT Biofilms are structured communities of tightly associated cells that constitute the predominant state of bacterial growth in natural and human-made environments. Although the core genetic circuitry that controls biofilm formation in model bacteria such as Bacillus subtilis has been well characterized, little is known about the role that metabolism plays in this complex developmental process. Here, we performed a time-resolved analysis of the metabolic changes associated with pellicle biofilm formation and development in B. subtilis by combining metabolomic, transcriptomic, and proteomic analyses. We report surprisingly widespread and dynamic remodeling of metabolism affecting central carbon metabolism, primary biosynthetic pathways, fermentation pathways, and secondary metabolism. Most of these metabolic alterations were hitherto unrecognized as biofilm associated. For example, we observed increased activity of the tricarboxylic acid (TCA) cycle during early biofilm growth, a shift from fatty acid biosynthesis to fatty acid degradation, reorganization of iron metabolism and transport, and a switch from acetate to acetoin fermentation. Close agreement between metabolomic, transcriptomic, and proteomic measurements indicated that remodeling of metabolism during biofilm development was largely controlled at the transcriptional level. Our results also provide insights into the transcription factors and regulatory networks involved in this complex metabolic remodeling. Following upon these results, we demonstrated that acetoin production via acetolactate synthase is essential for robust biofilm growth and has the dual role of conserving redox balance and maintaining extracellular pH. This report represents a comprehensive systems-level investigation of the metabolic remodeling occurring during B. subtilis biofilm development that will serve as a useful road map for future studies on biofilm physiology. IMPORTANCE Bacterial biofilms are ubiquitous in natural environments and play an important role in many clinical, industrial, and ecological settings. Although much is known about the transcriptional regulatory networks that control biofilm formation in model bacteria such as Bacillus subtilis , very little is known about the role of metabolism in this complex developmental process. To address this important knowledge gap, we performed a time-resolved analysis of the metabolic changes associated with bacterial biofilm development in B. subtilis by combining metabolomic, transcriptomic, and proteomic analyses. Here, we report a widespread and dynamic remodeling of metabolism affecting central carbon metabolism, primary biosynthetic pathways, fermentation pathways, and secondary metabolism. This report serves as a unique hypothesis-generating resource for future studies on bacterial biofilm physiology. Outside the biofilm research area, this work should also prove relevant to any investigators interested in microbial physiology and metabolism.
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Onggi’s permeability to carbon dioxide accelerates kimchi fermentation
Since ancient times, Korean chefs have fermented foods in an onggi, a traditional earthenware vessel. The porous structure of the onggi mimics the loose soil where lactic acid bacteria is naturally found. This permeability has been purported to facilitate the growth of lactic acid bacteria, but the details of the process remain poorly understood. In this combined experimental and theoretical study, we ferment salted napa cabbage in onggi and hermetic glassware and measure the time course of carbon dioxide concentration, which is a signature of fermentation. We present a mathematical model for carbon dioxide generation rate during fermentation using the onggi’s gas permeability as a free parameter. Our model provides a good fit for the data, and we conclude that porous walls help the onggi to ‘exhale’ carbon dioxide, lowering internal levels to those favoured by lactic acid bacteria. The positive pressure inside the onggi and the constant outflow through its walls act as a safety valve for bacteria growth by blocking the entry of external contaminants without mechanical components. We hope this study draws attention to the work of traditional artisans and inspires energy-efficient methods for fermenting and storing food products.
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- Award ID(s):
- 1806833
- PAR ID:
- 10459085
- Date Published:
- Journal Name:
- Journal of The Royal Society Interface
- Volume:
- 20
- Issue:
- 201
- ISSN:
- 1742-5662
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1098/rsif.2023.0034
