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Title: Controlled production of cellulases in plants for biomass conversion. Annual report, March 11, 1997--March 14, 1998

Abstract

The goal of this project is to facilitate conversion of plant biomass to usable energy by developing transgenic plants that express genes for microbial cellulases, which can be activated after harvest of the plants. In particular, the feasibility of targeting an endoglucanase and a cellobiohydrolase to the plant apoplast (cell wall milieu) is to be determined. To avoid detrimental effects of cellulose expression in plants, enzymes with high temperature optima were chosen; the genes for these enzymes are from thermophilic organisms that can use cellulose as a sole energy source. During the past year (year 2 of the grant), efforts have been focused on testing expression of endoglucanase E{sub 1}, from Acidothermus cellulolyticus, in the apoplast of both tobacco suspension cells and Arabidopsis thaliana plants. Using the plasmids constructed during the first year, transgenic cells and plants that contain the gene for the E{sub 1} catalytic domain fused to a signal peptide sequence were obtained. This gene was constructed so that the fusion protein will be secreted into the apoplast. The enzyme is made in large quantities and is secreted into the apoplast. More importantly, it is enzymatically active when placed under optimal reaction conditions (high temperature). Moreover, the plantmore » cells and intact plants exhibit no obvious problems with growth and development under laboratory conditions. Work has also continued to improve binary vectors for Agrobacterium-mediated transformation, to determine activity of E{sub 1} at various temperatures, and to investigate the activity of the 35S Cauliflower Mosaic Virus promoter in E. coli. 9 figs.« less

Authors:
Publication Date:
Research Org.:
Colorado Univ., Boulder, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Research, Washington, DC (United States)
OSTI Identifier:
621862
Report Number(s):
DOE/ER/12194-T2
ON: DE98003577; TRN: 98:002320
DOE Contract Number:  
FG03-96ER12194
Resource Type:
Technical Report
Resource Relation:
Other Information: PBD: [1998]
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; PROGRESS REPORT; GENETIC ENGINEERING; PLASMIDS; CELLULASE; ENZYME ACTIVITY; ARABIDOPSIS; TOBACCO; TRANSGENIC PLANTS

Citation Formats

Danna, K.J.. Controlled production of cellulases in plants for biomass conversion. Annual report, March 11, 1997--March 14, 1998. United States: N. p., 1998. Web. doi:10.2172/621862.
Danna, K.J.. Controlled production of cellulases in plants for biomass conversion. Annual report, March 11, 1997--March 14, 1998. United States. doi:10.2172/621862.
Danna, K.J.. Mon . "Controlled production of cellulases in plants for biomass conversion. Annual report, March 11, 1997--March 14, 1998". United States. doi:10.2172/621862. https://www.osti.gov/servlets/purl/621862.
@article{osti_621862,
title = {Controlled production of cellulases in plants for biomass conversion. Annual report, March 11, 1997--March 14, 1998},
author = {Danna, K.J.},
abstractNote = {The goal of this project is to facilitate conversion of plant biomass to usable energy by developing transgenic plants that express genes for microbial cellulases, which can be activated after harvest of the plants. In particular, the feasibility of targeting an endoglucanase and a cellobiohydrolase to the plant apoplast (cell wall milieu) is to be determined. To avoid detrimental effects of cellulose expression in plants, enzymes with high temperature optima were chosen; the genes for these enzymes are from thermophilic organisms that can use cellulose as a sole energy source. During the past year (year 2 of the grant), efforts have been focused on testing expression of endoglucanase E{sub 1}, from Acidothermus cellulolyticus, in the apoplast of both tobacco suspension cells and Arabidopsis thaliana plants. Using the plasmids constructed during the first year, transgenic cells and plants that contain the gene for the E{sub 1} catalytic domain fused to a signal peptide sequence were obtained. This gene was constructed so that the fusion protein will be secreted into the apoplast. The enzyme is made in large quantities and is secreted into the apoplast. More importantly, it is enzymatically active when placed under optimal reaction conditions (high temperature). Moreover, the plant cells and intact plants exhibit no obvious problems with growth and development under laboratory conditions. Work has also continued to improve binary vectors for Agrobacterium-mediated transformation, to determine activity of E{sub 1} at various temperatures, and to investigate the activity of the 35S Cauliflower Mosaic Virus promoter in E. coli. 9 figs.},
doi = {10.2172/621862},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Mon Jun 01 00:00:00 EDT 1998},
month = {Mon Jun 01 00:00:00 EDT 1998}
}

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