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Title: Custom fabrication of biomass containment devices using 3-D printing enables bacterial growth analyses with complex insoluble substrates

Abstract

Physiological studies of recalcitrant polysaccharide degradation are challenging for several reasons, one of which is the difficulty in obtaining a reproducibly accurate real-time measurement of bacterial growth using insoluble substrates. Current methods suffer from several problems including (i) high background noise due to the insoluble material interspersed with cells, (ii) high consumable and reagent cost and (iii) significant time delay between sampling and data acquisition. A customizable substrate and cell separation device would provide an option to study bacterial growth using optical density measurements. To test this hypothesis we used 3-D printing to create biomass containment devices that allow interaction between insoluble substrates and microbial cells but do not interfere with spectrophotometer measurements. Evaluation of materials available for 3-D printing indicated that UV-cured acrylic plastic was the best material, being superior to nylon or stainless steel when examined for heat tolerance, reactivity, and ability to be sterilized. Cost analysis of the 3-D printed devices indicated they are a competitive way to quantitate bacterial growth compared to viable cell counting or protein measurements, and experimental conditions were scalable over a 100-fold range. The presence of the devices did not alter growth phenotypes when using either soluble substrates or insoluble substrates. Furthermore,more » we applied biomass containment to characterize growth of Cellvibrio japonicus on authentic lignocellulose (non-pretreated corn stover), and found physiological evidence that xylan is a significant nutritional source despite an abundance of cellulose present.« less

Authors:
 [1];  [1]; ORCiD logo [1]
  1. Univ. of Maryland Baltimore County (UMBC), Baltimore, MD (United States)
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) (SC-23)
OSTI Identifier:
1326371
Alternate Identifier(s):
OSTI ID: 1359567
Grant/Contract Number:
SC0014183
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Microbiological Methods
Additional Journal Information:
Journal Volume: 130; Journal Issue: C; Journal ID: ISSN 0167-7012
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 3-D printing; biomass containment; Cellvibrio japonicus; lignocellulose; polysaccharide degradation

Citation Formats

Nelson, Cassandra E., Beri, Nina R., and Gardner, Jeffrey G. Custom fabrication of biomass containment devices using 3-D printing enables bacterial growth analyses with complex insoluble substrates. United States: N. p., 2016. Web. doi:10.1016/j.mimet.2016.09.013.
Nelson, Cassandra E., Beri, Nina R., & Gardner, Jeffrey G. Custom fabrication of biomass containment devices using 3-D printing enables bacterial growth analyses with complex insoluble substrates. United States. doi:10.1016/j.mimet.2016.09.013.
Nelson, Cassandra E., Beri, Nina R., and Gardner, Jeffrey G. Wed . "Custom fabrication of biomass containment devices using 3-D printing enables bacterial growth analyses with complex insoluble substrates". United States. doi:10.1016/j.mimet.2016.09.013. https://www.osti.gov/servlets/purl/1326371.
@article{osti_1326371,
title = {Custom fabrication of biomass containment devices using 3-D printing enables bacterial growth analyses with complex insoluble substrates},
author = {Nelson, Cassandra E. and Beri, Nina R. and Gardner, Jeffrey G.},
abstractNote = {Physiological studies of recalcitrant polysaccharide degradation are challenging for several reasons, one of which is the difficulty in obtaining a reproducibly accurate real-time measurement of bacterial growth using insoluble substrates. Current methods suffer from several problems including (i) high background noise due to the insoluble material interspersed with cells, (ii) high consumable and reagent cost and (iii) significant time delay between sampling and data acquisition. A customizable substrate and cell separation device would provide an option to study bacterial growth using optical density measurements. To test this hypothesis we used 3-D printing to create biomass containment devices that allow interaction between insoluble substrates and microbial cells but do not interfere with spectrophotometer measurements. Evaluation of materials available for 3-D printing indicated that UV-cured acrylic plastic was the best material, being superior to nylon or stainless steel when examined for heat tolerance, reactivity, and ability to be sterilized. Cost analysis of the 3-D printed devices indicated they are a competitive way to quantitate bacterial growth compared to viable cell counting or protein measurements, and experimental conditions were scalable over a 100-fold range. The presence of the devices did not alter growth phenotypes when using either soluble substrates or insoluble substrates. Furthermore, we applied biomass containment to characterize growth of Cellvibrio japonicus on authentic lignocellulose (non-pretreated corn stover), and found physiological evidence that xylan is a significant nutritional source despite an abundance of cellulose present.},
doi = {10.1016/j.mimet.2016.09.013},
journal = {Journal of Microbiological Methods},
number = C,
volume = 130,
place = {United States},
year = {Wed Sep 21 00:00:00 EDT 2016},
month = {Wed Sep 21 00:00:00 EDT 2016}
}

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