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Title: Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum

Abstract

Clostridium thermocellum parent ..delta..hpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentations when compared to the ..delta..hpt strain.

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1409731
Report Number(s):
NREL/JA-2700-70541
Journal ID: ISSN 0141-5492
DOE Contract Number:
AC36-08GO28308
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biotechnology Letters; Journal Volume: 39; Journal Issue: 551
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; bioethanol; consolidated bioprocessing; high throughput screening; lignocellulosic biomass; microplate assay

Citation Formats

Oguntimein, Gbekeloluwa B., Rodriguez, Miguel, Dumitrache, Alexandru, Shollenberger, Todd, Decker, Stephen R., Davison, Brian H., and Brown, Steven D. Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum. United States: N. p., 2017. Web. doi:10.1007/s10529-017-2467-2.
Oguntimein, Gbekeloluwa B., Rodriguez, Miguel, Dumitrache, Alexandru, Shollenberger, Todd, Decker, Stephen R., Davison, Brian H., & Brown, Steven D. Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum. United States. doi:10.1007/s10529-017-2467-2.
Oguntimein, Gbekeloluwa B., Rodriguez, Miguel, Dumitrache, Alexandru, Shollenberger, Todd, Decker, Stephen R., Davison, Brian H., and Brown, Steven D. 2017. "Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum". United States. doi:10.1007/s10529-017-2467-2.
@article{osti_1409731,
title = {Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum},
author = {Oguntimein, Gbekeloluwa B. and Rodriguez, Miguel and Dumitrache, Alexandru and Shollenberger, Todd and Decker, Stephen R. and Davison, Brian H. and Brown, Steven D.},
abstractNote = {Clostridium thermocellum parent ..delta..hpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentations when compared to the ..delta..hpt strain.},
doi = {10.1007/s10529-017-2467-2},
journal = {Biotechnology Letters},
number = 551,
volume = 39,
place = {United States},
year = 2017,
month =
}
  • Here, to develop and prototype a high-throughput microplate assay to assess anaerobic microorganisms and lignocellulosic biomasses in a rapid, cost-effective screen for consolidated bioprocessing potential. Clostridium thermocellum parent Δ hpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentationsmore » when compared to the Δ hpt strain. A prototype microplate assay system is developed that will facilitate high-throughput bioprospecting for new lignocellulosic biomass types, genetic variants and new microbial strains for bioethanol production.« less
  • Here, to develop and prototype a high-throughput microplate assay to assess anaerobic microorganisms and lignocellulosic biomasses in a rapid, cost-effective screen for consolidated bioprocessing potential. Clostridium thermocellum parent Δ hpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentationsmore » when compared to the Δ hpt strain. A prototype microplate assay system is developed that will facilitate high-throughput bioprospecting for new lignocellulosic biomass types, genetic variants and new microbial strains for bioethanol production.« less
  • Adsorption of Avicel-hydrolyzing activity was examined with respect to: mixed hardwood flour pretreated with 1% sulfuric acid for 9 s at 220C (PTW220), lignin prepared from PTW220 by either acid or enzymatic hydrolysis, and Avicel. Experiments were conducted at 60C for all materials, and also at 25C for PTW220. Based on transient adsorption results and reaction rates, times were selected at which to characterize adsorption at 60C as follows: PTW220, 1 min; lignin, 30 min; and Avicel, 45 min. Similar results were obtained for adsorption of cellulase activity to PTW220 at 25 and 60C, and for lignin prepared by enzymaticmore » and acid hydrolysis. For all materials, adsorption was described well by a Langmuir equation, although the reversibility of adsorption was not investigated. Langmuir affinity constants (L/g) were: PTW220, 109; lignin, 17.9; Avicel, 4.3; cellulose from PTW220, [ge]187. Langmuir capacity constants were 760 for PTW220 and 42 for Avicel; the cellulase binding capacity of lignin appeared to be very high under the conditions examined, and could not be determined. At low and moderate cellulase loadings at least, the majority of cellulase activity adsorbed to PTVV220 is bound to the cellulosic component. The results indicate that PTW220, and its cellulose component in particular, differ radically from Avicel with respect to adsorption. Avicel-hydrolyzing activity and CMC-hydrolyzing activities were found to bind to Avicel with a constant ratio of essentially one, consistent with adsorption of a multi-activity complex.« less