In planta expression of exocellulase enzymes for bio-ethanol production

Budathoki, P 2018, In planta expression of exocellulase enzymes for bio-ethanol production, Doctor of Philosophy (PhD), Science, RMIT University.

Document type: Thesis
Collection: Theses

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Title In planta expression of exocellulase enzymes for bio-ethanol production
Author(s) Budathoki, P
Year 2018
Abstract The use of bioethanol as an alternative fuel can help reduce greenhouse emissions associated with the consumption of crude petroleum oil and also slow the depletion of fossil fuel reserves. However, first generation bioethanol produced from starch or sugar crops cannot meet the enormous demand for fuel ethanol due to limited production capacity using available agricultural land and the ensuing competition for food and feed production. The production of ethanol from lignocellulosic crop biomass is challenging, but considering the availability of abundant quantities available globally, it has the potential to help meet the increasing requirement for clean and sustainable energy generation in the future. One of the major challenges in making cellulosic ethanol cost-competitive is to reduce the production of cellbiohydrolase (CBH) the predominant enzyme required for the degradation of biomass into simple sugars prior to fermentation. This project evaluated the expression of CBHI in the chloroplasts of tobacco, this was based on the premise that chloroplast based expression of other recombinant proteins have shown increased yield.Two genes encoding exocellulases; cbhI from Trichoderma reesei, and a novel gene CO14 (a protozoan cellulase isolated from termite gut) were selected for expression. There are no reports of these enzymes having been expressed in chloroplasts. This project generated transgenic tobacco lines that produced very limited amounts of recombinant exocellulases (CBHI/CO14) in the chloroplasts, however this expression generated a phenotype in that had major changes in the choloroplast ultrastructure; indicating that in plants the chloroplast is not a suitable organelle for transgenic expression of exocellulases.
Degree Doctor of Philosophy (PhD)
Institution RMIT University
School, Department or Centre Science
Subjects Industrial Biotechnology Diagnostics (incl. Biosensors)
Industrial Microbiology (incl. Biofeedstocks)
Industrial Molecular Engineering of Nucleic Acids and Proteins
Industrial Biotechnology not elsewhere classified
Microbial Genetics
Keyword(s) Cellbiohydrolase (CBH)
Trichoderma reesei
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Created: Tue, 11 Sep 2018, 14:43:51 EST by Adam Rivett
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