Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein.
2-methyl-1-butanol is a promising biofuel which could replace the world’s dependence on petroleum diesel. However, industrial production of this alcohol using biocatalyst is inefficient. The average cost of producing a unit of 2-methyl-1-butanol is more than that of petroleum diesel. Hence, better b...
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| Other Authors: | |
| Format: | Final Year Project (FYP) |
| Language: | English |
| Published: |
2010
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| Online Access: | http://hdl.handle.net/10356/39867 |
| _version_ | 1811684684432670720 |
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| author | Khoo, Choon Mong. |
| author2 | Jiang Rongrong |
| author_facet | Jiang Rongrong Khoo, Choon Mong. |
| author_sort | Khoo, Choon Mong. |
| collection | NTU |
| description | 2-methyl-1-butanol is a promising biofuel which could replace the world’s dependence on petroleum diesel. However, industrial production of this alcohol using biocatalyst is inefficient. The average cost of producing a unit of 2-methyl-1-butanol is more than that of petroleum diesel. Hence, better biocatalyst has to be engineered for better rate of production. To discover prokaryotic industrial strains with high tolerance for 2-methyl-1-butanol, CRP which is a well-investigated global transcription factor in Escherichia coli is mutated.
Error-prone PCR was administered on the crp gene which was then inserted into a pKSC101 plasmid. This recombinant plasmid was transformed into competent E. coli DH5 alpha cells. Subsequently, the transformed cells were investigated for the desired phenotype. This investigation was done based on the directed evolution concept.
Three better strains were discovered with their mutations determined. E. coli with shortened crp gene were found to be more resilient towards 2-methyl-1-butanol. However, there was no evidence suggesting how shortened crp gene was going to affect genetically engineered prokaryotic industrial strains production of the 2-methyl-1-butanol. |
| first_indexed | 2024-10-01T04:32:32Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/39867 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:32:32Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | ntu-10356/398672023-03-03T15:35:07Z Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein. Khoo, Choon Mong. Jiang Rongrong School of Chemical and Biomedical Engineering DRNTU::Engineering::Chemical engineering::Biotechnological production 2-methyl-1-butanol is a promising biofuel which could replace the world’s dependence on petroleum diesel. However, industrial production of this alcohol using biocatalyst is inefficient. The average cost of producing a unit of 2-methyl-1-butanol is more than that of petroleum diesel. Hence, better biocatalyst has to be engineered for better rate of production. To discover prokaryotic industrial strains with high tolerance for 2-methyl-1-butanol, CRP which is a well-investigated global transcription factor in Escherichia coli is mutated. Error-prone PCR was administered on the crp gene which was then inserted into a pKSC101 plasmid. This recombinant plasmid was transformed into competent E. coli DH5 alpha cells. Subsequently, the transformed cells were investigated for the desired phenotype. This investigation was done based on the directed evolution concept. Three better strains were discovered with their mutations determined. E. coli with shortened crp gene were found to be more resilient towards 2-methyl-1-butanol. However, there was no evidence suggesting how shortened crp gene was going to affect genetically engineered prokaryotic industrial strains production of the 2-methyl-1-butanol. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2010-06-07T06:14:09Z 2010-06-07T06:14:09Z 2010 2010 Final Year Project (FYP) http://hdl.handle.net/10356/39867 en Nanyang Technological University 58 p. application/pdf |
| spellingShingle | DRNTU::Engineering::Chemical engineering::Biotechnological production Khoo, Choon Mong. Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein. |
| title | Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein. |
| title_full | Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein. |
| title_fullStr | Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein. |
| title_full_unstemmed | Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein. |
| title_short | Increasing 2-methyl-1-butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein. |
| title_sort | increasing 2 methyl 1 butanol tolerance of prokaryotic industrial strains by using randomized libraries of camp receptor protein |
| topic | DRNTU::Engineering::Chemical engineering::Biotechnological production |
| url | http://hdl.handle.net/10356/39867 |
| work_keys_str_mv | AT khoochoonmong increasing2methyl1butanoltoleranceofprokaryoticindustrialstrainsbyusingrandomizedlibrariesofcampreceptorprotein |