Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae
Background: Hydrocarbon alkanes, components of major fossil fuels, are considered as next-generation biofuels because their biological production has recently been shown to be possible. However, high-yield alkane production requires robust host cells that are tolerant against alkanes, which exhibit...
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2013
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| Online Access: | https://hdl.handle.net/10356/96110 http://hdl.handle.net/10220/10083 |
| _version_ | 1811684423598342144 |
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| author | Chen, Binbin Ling, Hua Chang, Matthew Wook |
| author2 | School of Chemical and Biomedical Engineering |
| author_facet | School of Chemical and Biomedical Engineering Chen, Binbin Ling, Hua Chang, Matthew Wook |
| author_sort | Chen, Binbin |
| collection | NTU |
| description | Background: Hydrocarbon alkanes, components of major fossil fuels, are considered as next-generation biofuels because their biological production has recently been shown to be possible. However, high-yield alkane production requires robust host cells that are tolerant against alkanes, which exhibit cytotoxicity. In this study, we aimed to improve alkane tolerance in Saccharomyces cerevisiae, a key industrial microbial host, by harnessing heterologous transporters that potentially pump out alkanes. Results: To this end, we attempted to exploit ABC transporters in Yarrowia lipolytica based on the observation that it utilizes alkanes as a carbon source. We confirmed the increased transcription of ABC2 and ABC3 transporters upon exposure to a range of alkanes in Y. lipolytica. We then showed that the heterologous expression of ABC2 and ABC3 transporters significantly increased tolerance against decane and undecane in S. cerevisiae through maintaining lower intracellular alkane level. In particular, ABC2 transporter increased the tolerance limit of S. cerevisiae about 80-fold against decane. Furthermore, through site-directed mutagenesis for glutamate (E988 for ABC2, and E989 for ABC3) and histidine (H1020 for ABC2, and H1021 for ABC3), we provided the evidence that glutamate was essential for the activity of ABC2 and ABC3 transporters, with ATP most likely to be hydrolyzed by a catalytic carboxylate mechanism. Conclusions: Here, we demonstrated that transporter engineering through expression of heterologous efflux pumps led to significantly improved tolerance against alkane biofuels in S. cerevisiae. We believe that our results laid the groundwork for developing robust alkane-producing yeast cells through transporter engineering, which will greatly aid in next-generation alkane biofuel production and recovery. |
| first_indexed | 2024-10-01T04:28:24Z |
| format | Journal Article |
| id | ntu-10356/96110 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:28:24Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | ntu-10356/961102023-12-29T06:47:49Z Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae Chen, Binbin Ling, Hua Chang, Matthew Wook School of Chemical and Biomedical Engineering Background: Hydrocarbon alkanes, components of major fossil fuels, are considered as next-generation biofuels because their biological production has recently been shown to be possible. However, high-yield alkane production requires robust host cells that are tolerant against alkanes, which exhibit cytotoxicity. In this study, we aimed to improve alkane tolerance in Saccharomyces cerevisiae, a key industrial microbial host, by harnessing heterologous transporters that potentially pump out alkanes. Results: To this end, we attempted to exploit ABC transporters in Yarrowia lipolytica based on the observation that it utilizes alkanes as a carbon source. We confirmed the increased transcription of ABC2 and ABC3 transporters upon exposure to a range of alkanes in Y. lipolytica. We then showed that the heterologous expression of ABC2 and ABC3 transporters significantly increased tolerance against decane and undecane in S. cerevisiae through maintaining lower intracellular alkane level. In particular, ABC2 transporter increased the tolerance limit of S. cerevisiae about 80-fold against decane. Furthermore, through site-directed mutagenesis for glutamate (E988 for ABC2, and E989 for ABC3) and histidine (H1020 for ABC2, and H1021 for ABC3), we provided the evidence that glutamate was essential for the activity of ABC2 and ABC3 transporters, with ATP most likely to be hydrolyzed by a catalytic carboxylate mechanism. Conclusions: Here, we demonstrated that transporter engineering through expression of heterologous efflux pumps led to significantly improved tolerance against alkane biofuels in S. cerevisiae. We believe that our results laid the groundwork for developing robust alkane-producing yeast cells through transporter engineering, which will greatly aid in next-generation alkane biofuel production and recovery. Published version 2013-06-10T01:50:54Z 2019-12-06T19:25:55Z 2013-06-10T01:50:54Z 2019-12-06T19:25:55Z 2013 2013 Journal Article Chen, B., Ling, H., & Chang, M. W. (2013). Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae. Biotechnology for Biofuels, 6(21). https://hdl.handle.net/10356/96110 http://hdl.handle.net/10220/10083 10.1186/1754-6834-6-21 23402697 en Biotechnology for biofuels © 2013 Chen et al. This paper was published in Biotechnology for Biofuels and is made available as an electronic reprint (preprint) with permission of Chen et al. The paper can be found at the following official DOI: [http://dx.doi.org/10.1186/1754-6834-6-21]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law. application/pdf |
| spellingShingle | Chen, Binbin Ling, Hua Chang, Matthew Wook Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae |
| title | Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae |
| title_full | Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae |
| title_fullStr | Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae |
| title_full_unstemmed | Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae |
| title_short | Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae |
| title_sort | transporter engineering for improved tolerance against alkane biofuels in saccharomyces cerevisiae |
| url | https://hdl.handle.net/10356/96110 http://hdl.handle.net/10220/10083 |
| work_keys_str_mv | AT chenbinbin transporterengineeringforimprovedtoleranceagainstalkanebiofuelsinsaccharomycescerevisiae AT linghua transporterengineeringforimprovedtoleranceagainstalkanebiofuelsinsaccharomycescerevisiae AT changmatthewwook transporterengineeringforimprovedtoleranceagainstalkanebiofuelsinsaccharomycescerevisiae |