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Title: Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions

Abstract

Microbial communities enriched from diverse environments have shown considerable promise for the targeted discovery of microorganisms and enzymes for bioconversion of lignocellulose to liquid fuels. While preservation of microbial communities is important for commercialization and research, few studies have examined storage conditions ideal for preservation. The goal of this study was to evaluate the impact of preservation method on composition of microbial communities enriched on switchgrass before and after storage. The enrichments were completed in a high-solid and aerobic environment at 55 °C. Community composition was examined for each enrichment to determine when a stable community was achieved. Preservation methods included cryopreservation with the cryoprotective agents DMSO and glycerol, and cryopreservation without cryoprotective agents. Revived communities were examined for their ability to decompose switchgrass under high-solid and thermophilic conditions. High-throughput 16S rRNA gene sequencing of DNA extracted from enrichment samples showed that the majority of the shift in composition of the switchgrass-degrading community occurred during the initial three 2-week enrichments. Shifts in community structure upon storage occurred in all cryopreserved samples. Storage in liquid nitrogen in the absence of cryoprotectant resulted in variable preservation of dominant microorganisms in enriched samples. Cryopreservation with either DMSO or glycerol provided consistent and equivalentmore » preservation of dominant organisms. In conclusion, a stable switchgrass-degrading microbial community was achieved after three 2-week enrichments. Dominant microorganisms were preserved equally well with DMSO and glycerol. DMSO-preserved communities required more incubation time upon revival to achieve pre-storage activity levels during high-solid thermophilic cultivation on switchgrass. Despite shifts in the community with storage, the samples were active upon revival under thermophilic and high-solid conditions. The results suggest that the presence of microorganisms may be more important than their relative abundance in retaining an active microbial community.« less

Authors:
 [1];  [2];  [3];  [4];  [5]; ORCiD logo [2]
  1. Univ. of California, Davis, CA (United States)
  2. Univ. of California, Davis, CA (United States); Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  3. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Davis, CA (United States). Dept. of Food Science and Technology
  4. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
  5. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1257272
Grant/Contract Number:  
CA-D-BAE-2228-RR; 12-LR-237496; AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; biological lignocellulose deconstruction; cryopreservation; microbial community enrichment

Citation Formats

Yu, Chaowei, Reddy, Amitha P., Simmons, Christopher W., Simmons, Blake A., Singer, Steven W., and VanderGheynst, Jean S. Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions. United States: N. p., 2015. Web. doi:10.1186/s13068-015-0392-y.
Yu, Chaowei, Reddy, Amitha P., Simmons, Christopher W., Simmons, Blake A., Singer, Steven W., & VanderGheynst, Jean S. Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions. United States. doi:10.1186/s13068-015-0392-y.
Yu, Chaowei, Reddy, Amitha P., Simmons, Christopher W., Simmons, Blake A., Singer, Steven W., and VanderGheynst, Jean S. Wed . "Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions". United States. doi:10.1186/s13068-015-0392-y. https://www.osti.gov/servlets/purl/1257272.
@article{osti_1257272,
title = {Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions},
author = {Yu, Chaowei and Reddy, Amitha P. and Simmons, Christopher W. and Simmons, Blake A. and Singer, Steven W. and VanderGheynst, Jean S.},
abstractNote = {Microbial communities enriched from diverse environments have shown considerable promise for the targeted discovery of microorganisms and enzymes for bioconversion of lignocellulose to liquid fuels. While preservation of microbial communities is important for commercialization and research, few studies have examined storage conditions ideal for preservation. The goal of this study was to evaluate the impact of preservation method on composition of microbial communities enriched on switchgrass before and after storage. The enrichments were completed in a high-solid and aerobic environment at 55 °C. Community composition was examined for each enrichment to determine when a stable community was achieved. Preservation methods included cryopreservation with the cryoprotective agents DMSO and glycerol, and cryopreservation without cryoprotective agents. Revived communities were examined for their ability to decompose switchgrass under high-solid and thermophilic conditions. High-throughput 16S rRNA gene sequencing of DNA extracted from enrichment samples showed that the majority of the shift in composition of the switchgrass-degrading community occurred during the initial three 2-week enrichments. Shifts in community structure upon storage occurred in all cryopreserved samples. Storage in liquid nitrogen in the absence of cryoprotectant resulted in variable preservation of dominant microorganisms in enriched samples. Cryopreservation with either DMSO or glycerol provided consistent and equivalent preservation of dominant organisms. In conclusion, a stable switchgrass-degrading microbial community was achieved after three 2-week enrichments. Dominant microorganisms were preserved equally well with DMSO and glycerol. DMSO-preserved communities required more incubation time upon revival to achieve pre-storage activity levels during high-solid thermophilic cultivation on switchgrass. Despite shifts in the community with storage, the samples were active upon revival under thermophilic and high-solid conditions. The results suggest that the presence of microorganisms may be more important than their relative abundance in retaining an active microbial community.},
doi = {10.1186/s13068-015-0392-y},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 8,
place = {United States},
year = {2015},
month = {12}
}

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