Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitors
Abstract Background Furfural and acetic acid are the two major inhibitors generated during lignocellulose pretreatment and hydrolysis, would severely inhibit the cell growth, metabolism, and ethanol fermentation efficiency of Zymomonas mobilis. Effective genome shuffling mediated by protoplast elect...
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| Format: | Article |
| Language: | English |
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BMC
2019-12-01
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| Series: | Biotechnology for Biofuels |
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| Online Access: | https://doi.org/10.1186/s13068-019-1631-4 |
| _version_ | 1811231439210938368 |
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| author | Weiting Wang Bo Wu Han Qin Panting Liu Yao Qin Guowei Duan Guoquan Hu Mingxiong He |
| author_facet | Weiting Wang Bo Wu Han Qin Panting Liu Yao Qin Guowei Duan Guoquan Hu Mingxiong He |
| author_sort | Weiting Wang |
| collection | DOAJ |
| description | Abstract Background Furfural and acetic acid are the two major inhibitors generated during lignocellulose pretreatment and hydrolysis, would severely inhibit the cell growth, metabolism, and ethanol fermentation efficiency of Zymomonas mobilis. Effective genome shuffling mediated by protoplast electrofusion was developed and then applied to Z. mobilis. Results After two rounds of genome shuffling, 10 different mutants with improved cell growth and ethanol yield in the presence of 5.0 g/L acetic acid and 3.0 g/L furfural were obtained. The two most prominent genome-shuffled strains, 532 and 533, were further investigated along with parental strains in the presence of 7.0 g/L acetic acid and 3.0 g/L furfural. The results showed that mutants 532 and 533 were superior to the parental strain AQ8-1 in the presence of 7.0 g/L acetic acid, with a shorter fermentation time (30 h) and higher productivity than AQ8-1. Mutant 533 exhibited subtle differences from parental strain F34 in the presence of 3.0 g/L furfural. Mutations present in 10 genome-shuffled strains were identified via whole-genome resequencing, and the source of each mutation was identified as either de novo mutation or recombination of the parent genes. Conclusions These results indicate that genome shuffling is an efficient method for enhancing stress tolerance in Z. mobilis. The engineered strains generated in this study could be potential cellulosic ethanol producers in the future. |
| first_indexed | 2024-04-12T10:45:21Z |
| format | Article |
| id | doaj.art-0aae7fe98bfb482f8e4f0ca579a94802 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-04-12T10:45:21Z |
| publishDate | 2019-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-0aae7fe98bfb482f8e4f0ca579a948022022-12-22T03:36:26ZengBMCBiotechnology for Biofuels1754-68342019-12-0112111210.1186/s13068-019-1631-4Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitorsWeiting Wang0Bo Wu1Han Qin2Panting Liu3Yao Qin4Guowei Duan5Guoquan Hu6Mingxiong He7Biomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsBiomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsBiomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsBiomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsBiomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsBiomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsBiomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsBiomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural AffairsAbstract Background Furfural and acetic acid are the two major inhibitors generated during lignocellulose pretreatment and hydrolysis, would severely inhibit the cell growth, metabolism, and ethanol fermentation efficiency of Zymomonas mobilis. Effective genome shuffling mediated by protoplast electrofusion was developed and then applied to Z. mobilis. Results After two rounds of genome shuffling, 10 different mutants with improved cell growth and ethanol yield in the presence of 5.0 g/L acetic acid and 3.0 g/L furfural were obtained. The two most prominent genome-shuffled strains, 532 and 533, were further investigated along with parental strains in the presence of 7.0 g/L acetic acid and 3.0 g/L furfural. The results showed that mutants 532 and 533 were superior to the parental strain AQ8-1 in the presence of 7.0 g/L acetic acid, with a shorter fermentation time (30 h) and higher productivity than AQ8-1. Mutant 533 exhibited subtle differences from parental strain F34 in the presence of 3.0 g/L furfural. Mutations present in 10 genome-shuffled strains were identified via whole-genome resequencing, and the source of each mutation was identified as either de novo mutation or recombination of the parent genes. Conclusions These results indicate that genome shuffling is an efficient method for enhancing stress tolerance in Z. mobilis. The engineered strains generated in this study could be potential cellulosic ethanol producers in the future.https://doi.org/10.1186/s13068-019-1631-4ElectrofusionGenome shufflingMutagenesisInhibitor toleranceZymomonas mobilis |
| spellingShingle | Weiting Wang Bo Wu Han Qin Panting Liu Yao Qin Guowei Duan Guoquan Hu Mingxiong He Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitors Biotechnology for Biofuels Electrofusion Genome shuffling Mutagenesis Inhibitor tolerance Zymomonas mobilis |
| title | Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitors |
| title_full | Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitors |
| title_fullStr | Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitors |
| title_full_unstemmed | Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitors |
| title_short | Genome shuffling enhances stress tolerance of Zymomonas mobilis to two inhibitors |
| title_sort | genome shuffling enhances stress tolerance of zymomonas mobilis to two inhibitors |
| topic | Electrofusion Genome shuffling Mutagenesis Inhibitor tolerance Zymomonas mobilis |
| url | https://doi.org/10.1186/s13068-019-1631-4 |
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