High-throughput screening of environmental polysaccharide-degrading bacteria using biomass containment and complex insoluble substrates

• Published in: Applied microbiology and biotechnology Vol. 104; no. 8; pp. 3379 - 3389
• Main Authors: Monge, Estela C., Levi, Marios, Forbin, Joseph N., Legesse, Mussie D., Udo, Basil A., deCarvalho, Tagide N., Gardner, Jeffrey G.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.04.2020; Springer; Springer Nature B.V
• Subjects: 3D printing; Bacteria; Bacteria - growth & development; Bacteria - isolation & purification; Bacteria - metabolism; BASIC BIOLOGICAL SCIENCES; Biodegradation; Biomass; Biomedical and Life Sciences; Biotechnological Products and Process Engineering; Biotechnology; Biotechnology - instrumentation; Biotechnology - methods; Carbohydrate Metabolism; Carbohydrates; Cellulose; Cellvibrio japonicus; Chitin; Containment; Degradation; Devices; Enzymes; Filtration; High-throughput screening; High-Throughput Screening Assays - instrumentation; High-Throughput Screening Assays - methods; Human nutrition; Life Sciences; Lignin - metabolism; Lignocellulose; Microbial Genetics and Genomics; Microbiology; Nutrient cycles; Nutrition; Physiological aspects; Polysaccharide; Polysaccharides; Polysaccharides - metabolism; Printing, Three-Dimensional; Saccharides; Screening; Solubility; Substrates; Three dimensional printing; Chitin; Lignocellulose; Polysaccharide; 3D printing; Cellulose; Cellvibrio japonicus
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-020-10469-3

Carbohydrate degradation by microbes plays an important role in global nutrient cycling, human nutrition, and biotechnological applications. Studies that focus on the degradation of complex recalcitrant polysaccharides are challenging because of the insolubility of these substrates as found in their natural contexts. Specifically, current methods to examine carbohydrate-based biomass degradation using bacterial strains or purified enzymes are not compatible with high-throughput screening using complex insoluble materials. In this report, we developed a small 3D printed filter device that fits inside a microplate well that allows for the free movement of bacterial cells, media, and enzymes while containing insoluble biomass. These devices do not interfere with standard microplate readers and can be used for both short- (24–48 h) and long-duration (> 100 h) experiments using complex insoluble substrates. These devices were used to quantitatively screen in a high-throughput manner environmental isolates for their ability to grow using lignocellulose or rice grains as a sole nutrient source. Additionally, we determined that the microplate-based containment devices are compatible with existing enzymatic assays to measure activity against insoluble biomass. Overall, these microplate containment devices provide a platform to study the degradation of complex insoluble materials in a high-throughput manner and have the potential to help uncover ecologically important aspects of bacterial metabolism as well as to accelerate biotechnological innovation.