Expression and Characterization of Acidothermus celluloyticus E1 Endoglucanase in Transgenic Duckweed Lemna minor 8627

Sun, Ye; Himmel, Michael E.; Skory, Christopher D.; Adney, William S.; Thomas, Steven R.; Tisserat, Brent; Nishimura, Yufuko; Yamamoto, Yuri T.

doi:10.1016/j.biortech.2006.09.055

Expression and Characterization of Acidothermus celluloyticus E1 Endoglucanase in Transgenic Duckweed Lemna minor 8627

Journal Article · Sun Dec 31 19:00:00 EST 2006 · Bioresource Technology

DOI:https://doi.org/10.1016/j.biortech.2006.09.055· OSTI ID:944493

Sun, Ye; Himmel, Michael E.; Skory, Christopher D.; Adney, William S.; Thomas, Steven R.; Tisserat, Brent; Nishimura, Yufuko; Yamamoto, Yuri T.

Endoglucanase E1 from Acidothermus cellulolyticus was expressed cytosolically under control of the cauliflower mosaic virus 35S promoter in transgenic duckweed, Lemna minor 8627 without any obvious observable phenotypic effects on morphology or rate of growth. The recombinant enzyme co-migrated with the purified catalytic domain fraction of the native E1 protein on western blot analysis, revealing that the cellulose-binding domain was cleaved near or in the linker region. The duckweed-expressed enzyme was biologically active and the expression level was up to 0.24% of total soluble protein. The endoglucanase activity with carboxymethylcellulose averaged 0.2 units mg protein-1 extracted from fresh duckweed. The optimal temperature and pH for E1 enzyme activity were about 80 degrees C and pH 5, respectively. While extraction with HEPES (N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid]) buffer (pH 8) resulted in the highest recovery of total soluble proteins and E1 enzyme, extraction with citrate buffer (pH 4.8) at 65 degrees C enriched relative amounts of E1 enzyme in the extract. This study demonstrates that duckweed may offer new options for the expression of cellulolytic enzymes in transgenic plants.

Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE

DOE Contract Number:: AC36-08GO28308

OSTI ID:: 944493

Report Number(s):: NREL/JA-270-41767; MainId:63494; UUID:56bcb3d3-fad4-e411-b769-d89d67132a6d; MainAdminId:45568

Journal Information:: Bioresource Technology, Journal Name: Bioresource Technology Journal Issue: 2007 Vol. 98; ISSN 0960-8524; ISSN BIRTEB

Country of Publication:: United States

Language:: English

References (15)

Characterisation of cellulose-hydrolysing enzymes from the fungusBipolaris sorokiniana Geimba, Mercedes P.; Riffel, Alessandro; Agostini, Viviane Journal of the Science of Food and Agriculture, Vol. 79, Issue 13 https://doi.org/10.1002/(SICI)1097-0010(199910)79:13<1849::AID-JSFA445>3.0.CO;2-4	journal	October 1999
Quantitation ofAcidothermus cellulolyticus E1 endoglucanase andThermomonospora fusca E3 exoglucanase using enzyme-linked immunosorbent assay (ELISA) Nieves, Rafael A.; Chou, Yat-Chen; Himmel, Michael E. Applied Biochemistry and Biotechnology, Vol. 51-52, Issue 1, p. 211-223 https://doi.org/10.1007/BF02933425	journal	September 1995
A new thermostable endoglucanase,Acidothermus cellulolyticus E1: Synergism withTrichoderma reesei CBH I and comparison to Thermomonospora fusca E₅ Baker, John O.; Adney, William S.; Nleves, Rafael A. Applied Biochemistry and Biotechnology, Vol. 45-46, Issue 1, p. 245-256 https://doi.org/10.1007/BF02941803	journal	March 1994
Genetic transformation of duckweed Lemna gibba and Lemna minor Yamamoto, Yuri T.; Rajbhandari, Nirmala; Lin, Xiaohong In Vitro Cellular & Developmental Biology - Plant, Vol. 37, Issue 3, p. 349-353 https://doi.org/10.1007/s11627-001-0062-6	journal	May 2001
Cellulase assay methods: a review Sharrock, Keith R. Journal of Biochemical and Biophysical Methods, Vol. 17, Issue 2 https://doi.org/10.1016/0165-022X(88)90040-1	journal	October 1988
Characterization and affinity applications of cellulose-binding domains Tomme, Peter; Boraston, Al; McLean, Brad Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 715, Issue 1 https://doi.org/10.1016/S0378-4347(98)00053-X	journal	September 1998
Nutrient recovery from swine lagoon water by Spirodela punctata Cheng, Jiayang; Bergmann, Ben A.; Classen, John J. Bioresource Technology, Vol. 81, Issue 1 https://doi.org/10.1016/S0960-8524(01)00098-0	journal	January 2002
In vitro selection of duckweed geographical isolates for potential use in swine lagoon effluent renovation Bergmann, B. A.; Cheng, J.; Classen, J. Bioresource Technology, Vol. 73, Issue 1 https://doi.org/10.1016/S0960-8524(99)00137-6	journal	May 2000
Outlook for cellulase improvement: Screening and selection strategies Percival Zhang, Y.-H.; Himmel, Michael E.; Mielenz, Jonathan R. Biotechnology Advances, Vol. 24, Issue 5, p. 452-481 https://doi.org/10.1016/j.biotechadv.2006.03.003	journal	September 2006
Crystal Structure of Thermostable Family 5 Endocellulase E1 from Acidothermus cellulolyticus in Complex with Cellotetraose ^† ^, ^‡ Sakon, Joshua; Adney, William S.; Himmel, Michael E. Biochemistry, Vol. 35, Issue 33 https://doi.org/10.1021/bi9604439	journal	January 1996
Improved plant-based production of E1 endoglucanase using potato: expression optimization and tissue targeting Dai, Ziyu; Hooker, Brian S.; Anderson, Daniel B. Molecular Breeding, Vol. 6, Issue 3, p. 277-285 https://doi.org/10.1023/A:1009653011948	journal	May 2000
Accumulation of a thermostable endo-1,4-β-D-glucanase in the apoplast of Arabidopsis thaliana leaves Ziegler, Matthew T.; Thomas, Steven R.; Danna, Kathleen J. Molecular Breeding, Vol. 6, Issue 1, p. 37-46 https://doi.org/10.1023/A:1009667524690	journal	January 2000
Dramatic effects of truncation and sub-cellular targeting on the accumulation of recombinant microbial cellulase in tobacco Ziegelhoffer, Thomas; Raasch, John A.; Austin-Phillips, Sandra Molecular Breeding, Vol. 8, Issue 2, p. 147-158 https://doi.org/10.1023/A:1013338312948	journal	September 2001
Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures Schenk, Roy U.; Hildebrandt, A. C. Canadian Journal of Botany, Vol. 50, Issue 1, p. 199-204 https://doi.org/10.1139/b72-026	journal	January 1972
Measurement of cellulase activities Ghose, T. K. Pure and Applied Chemistry, Vol. 59, Issue 2 https://doi.org/10.1351/pac198759020257	journal	January 1987

Similar Records

Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting

Journal Article · Thu Jun 01 00:00:00 EDT 2000 · Molecular Breeding · OSTI ID:15001181

Optimization of Acidothermus Celluloyticus Endoglucanase (E1) Production in Transgenic Tobacco Plants by Transcriptional, Post-transcription and Post-Translational Modification

Journal Article · Sat Oct 01 00:00:00 EDT 2005 · Transgenic Research, 14(5):627-643 · OSTI ID:860942

Gene coding for the E1 endoglucanase

Patent · Sun Dec 31 23:00:00 EST 1995 · OSTI ID:870513

Related Subjects

Acidothermus cellulolyticus
BASIC BIOLOGICAL SCIENCES
BIOMASS FUELS
BRASSICA
BUFFERS
CITRATES
ENZYME ACTIVITY
ENZYMES
Endoglucanase E1
Lemna minor
MORPHOLOGY
PROMOTERS
PROTEINS
TRANSGENIC PLANTS
gene expression
transgenic duckweed

Expression and Characterization of Acidothermus celluloyticus E1 Endoglucanase in Transgenic Duckweed Lemna minor 8627

Citation Formats

References (15)

Similar Records

Related Subjects