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

Abstract

Abstract 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 uncovermore » ecologically important aspects of bacterial metabolism as well as to accelerate biotechnological innovation.« less

Authors:
; ; ; ; ; ; ORCiD logo
Publication Date:
Research Org.:
Univ. of Maryland Baltimore County (UMBC), Baltimore, MD (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH)
OSTI Identifier:
1619313
Alternate Identifier(s):
OSTI ID: 1602749
Grant/Contract Number:  
SC0014183
Resource Type:
Published Article
Journal Name:
Applied Microbiology and Biotechnology
Additional Journal Information:
Journal Name: Applied Microbiology and Biotechnology Journal Volume: 104 Journal Issue: 8; Journal ID: ISSN 0175-7598
Publisher:
Springer Science + Business Media
Country of Publication:
Germany
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 3D printing; Cellvibrio japonicus; Cellulose; Chitin; Lignocellulose; Polysaccharide

Citation Formats

Monge, Estela C., Levi, Marios, Forbin, Joseph N., Legesse, Mussie D., Udo, Basil A., deCarvalho, Tagide N., and Gardner, Jeffrey G. High-throughput screening of environmental polysaccharide-degrading bacteria using biomass containment and complex insoluble substrates. Germany: N. p., 2020. Web. doi:10.1007/s00253-020-10469-3.
Monge, Estela C., Levi, Marios, Forbin, Joseph N., Legesse, Mussie D., Udo, Basil A., deCarvalho, Tagide N., & Gardner, Jeffrey G. High-throughput screening of environmental polysaccharide-degrading bacteria using biomass containment and complex insoluble substrates. Germany. doi:https://doi.org/10.1007/s00253-020-10469-3
Monge, Estela C., Levi, Marios, Forbin, Joseph N., Legesse, Mussie D., Udo, Basil A., deCarvalho, Tagide N., and Gardner, Jeffrey G. Sat . "High-throughput screening of environmental polysaccharide-degrading bacteria using biomass containment and complex insoluble substrates". Germany. doi:https://doi.org/10.1007/s00253-020-10469-3.
@article{osti_1619313,
title = {High-throughput screening of environmental polysaccharide-degrading bacteria using biomass containment and complex insoluble substrates},
author = {Monge, Estela C. and Levi, Marios and Forbin, Joseph N. and Legesse, Mussie D. and Udo, Basil A. and deCarvalho, Tagide N. and Gardner, Jeffrey G.},
abstractNote = {Abstract 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.},
doi = {10.1007/s00253-020-10469-3},
journal = {Applied Microbiology and Biotechnology},
number = 8,
volume = 104,
place = {Germany},
year = {2020},
month = {2}
}

Journal Article:
Free Publicly Available Full Text
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DOI: https://doi.org/10.1007/s00253-020-10469-3

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