Department of Chemistry, University of Washington, Box 351700, Seattle, WA, USA.
Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA.
Nat Commun. 2020 Feb 4;11(1):563. doi: 10.1038/s41467-020-14371-4.
Most mono- and co-culture bioprocess applications rely on large-scale suspension fermentation technologies that are not easily portable, reusable, or suitable for on-demand production. Here, we describe a hydrogel system for harnessing the bioactivity of embedded microbes for on-demand small molecule and peptide production in microbial mono-culture and consortia. This platform bypasses the challenges of engineering a multi-organism consortia by utilizing a temperature-responsive, shear-thinning hydrogel to compartmentalize organisms into polymeric hydrogels that control the final consortium composition and dynamics without the need for synthetic control of mutualism. We demonstrate that these hydrogels provide protection from preservation techniques (including lyophilization) and can sustain metabolic function for over 1 year of repeated use. This approach was utilized for the production of four chemical compounds, a peptide antibiotic, and carbohydrate catabolism by using either mono-cultures or co-cultures. The printed microbe-laden hydrogel constructs' efficiency in repeated production phases, both pre- and post-preservation, outperforms liquid culture.
大多数单培养物和共培养物生物工艺应用都依赖于大规模悬浮发酵技术，这些技术不易携带、重复使用或适用于按需生产。在这里，我们描述了一种水凝胶系统，用于利用嵌入式微生物的生物活性，在微生物单培养物和群落中按需生产小分子和肽。该平台通过利用温度响应、剪切稀化水凝胶将生物体分隔到聚合物水凝胶中来绕过工程多生物体群落的挑战，而无需对共生关系进行合成控制即可控制最终群落的组成和动态。我们证明，这些水凝胶可以防止保存技术（包括冻干）的影响，并可以维持代谢功能超过 1 年的重复使用。该方法用于使用单培养物或共培养物生产四种化学化合物、一种肽抗生素和碳水化合物分解代谢。与液体培养相比，打印的含有微生物的水凝胶构建体在保存前后的重复生产阶段的效率更高。
