Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic Hydrolysate
Bio-based xylonic acid produced from inexpensive lignocellulosic biomass has enormous market potential and enhances the overall economic benefits of biorefinery processes. In this study, the introduction of genes encoding xylose dehydrogenase driven by the promoter P<i>pdc</i> into <i...
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MDPI AG
2025-03-01
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| Series: | Fermentation |
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| author | Banrui Ruan Xiongying Yan Zhaoqing He Qiaoning He Shihui Yang |
| author_facet | Banrui Ruan Xiongying Yan Zhaoqing He Qiaoning He Shihui Yang |
| author_sort | Banrui Ruan |
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| description | Bio-based xylonic acid produced from inexpensive lignocellulosic biomass has enormous market potential and enhances the overall economic benefits of biorefinery processes. In this study, the introduction of genes encoding xylose dehydrogenase driven by the promoter P<i>pdc</i> into <i>Z. mobilis</i> using a plasmid vector resulted in the accumulation of xylonic acid at a titer of 16.8 ± 1.6 g/L. To achieve stable xylonic acid production, a gene cassette for xylonic acid production was integrated into the genome at the chromosomal locus of <i>ZMO0038</i> and <i>ZMO1650</i> using the endogenous type I-F CRISPR-Cas system. The titer of the resulting recombinant strain XA3 reduced to 12.2 ± 0.56 g/L, which could be the copy number difference between the plasmid and chromosomal integration. Oxygen content was then identified to be the key factor for xylonic acid production. To further increase xylonic acid production capability, a recombinant strain, XA9, with five copies of a gene cassette for xylonic acid production was constructed by integrating the gene cassette into the genome at the chromosomal locus of <i>ZMO1094</i>, <i>ZMO1547</i>, and <i>ZMO1577</i> on the basis of XA3. The titer of xylonic acid increased to 51.9 ± 0.1 g/L with a maximum yield of 1.10 g/g, which is close to the theoretical yield in a pure sugar medium. In addition, the recombinant strain XA9 is genetically stable and can produce 16.2 ± 0.14 g/L of xylonic acid with a yield of 1.03 ± 0.01 g/g in the lignocellulosic hydrolysate. Our study thus constructed a recombinant strain, XA9, of <i>Z. mobilis</i> for xylonic acid production from lignocellulosic hydrolysate, demonstrating the capability of <i>Z. mobilis</i> as a biorefinery chassis for economic lignocellulosic biochemical production. |
| format | Article |
| id | doaj-art-740962234e604e7db21e55c19395fd28 |
| institution | Kabale University |
| issn | 2311-5637 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj-art-740962234e604e7db21e55c19395fd282025-08-20T03:43:33ZengMDPI AGFermentation2311-56372025-03-0111314110.3390/fermentation11030141Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic HydrolysateBanrui Ruan0Xiongying Yan1Zhaoqing He2Qiaoning He3Shihui Yang4State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, ChinaBio-based xylonic acid produced from inexpensive lignocellulosic biomass has enormous market potential and enhances the overall economic benefits of biorefinery processes. In this study, the introduction of genes encoding xylose dehydrogenase driven by the promoter P<i>pdc</i> into <i>Z. mobilis</i> using a plasmid vector resulted in the accumulation of xylonic acid at a titer of 16.8 ± 1.6 g/L. To achieve stable xylonic acid production, a gene cassette for xylonic acid production was integrated into the genome at the chromosomal locus of <i>ZMO0038</i> and <i>ZMO1650</i> using the endogenous type I-F CRISPR-Cas system. The titer of the resulting recombinant strain XA3 reduced to 12.2 ± 0.56 g/L, which could be the copy number difference between the plasmid and chromosomal integration. Oxygen content was then identified to be the key factor for xylonic acid production. To further increase xylonic acid production capability, a recombinant strain, XA9, with five copies of a gene cassette for xylonic acid production was constructed by integrating the gene cassette into the genome at the chromosomal locus of <i>ZMO1094</i>, <i>ZMO1547</i>, and <i>ZMO1577</i> on the basis of XA3. The titer of xylonic acid increased to 51.9 ± 0.1 g/L with a maximum yield of 1.10 g/g, which is close to the theoretical yield in a pure sugar medium. In addition, the recombinant strain XA9 is genetically stable and can produce 16.2 ± 0.14 g/L of xylonic acid with a yield of 1.03 ± 0.01 g/g in the lignocellulosic hydrolysate. Our study thus constructed a recombinant strain, XA9, of <i>Z. mobilis</i> for xylonic acid production from lignocellulosic hydrolysate, demonstrating the capability of <i>Z. mobilis</i> as a biorefinery chassis for economic lignocellulosic biochemical production.https://www.mdpi.com/2311-5637/11/3/141xylonic acidxylose dehydrogenase<i>Zymomonas mobilis</i>metabolic engineeringlignocellulosic hydrolysate |
| spellingShingle | Banrui Ruan Xiongying Yan Zhaoqing He Qiaoning He Shihui Yang Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic Hydrolysate Fermentation xylonic acid xylose dehydrogenase <i>Zymomonas mobilis</i> metabolic engineering lignocellulosic hydrolysate |
| title | Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic Hydrolysate |
| title_full | Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic Hydrolysate |
| title_fullStr | Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic Hydrolysate |
| title_full_unstemmed | Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic Hydrolysate |
| title_short | Metabolic Engineering of <i>Zymomonas mobilis</i> for Xylonic Acid Production from Lignocellulosic Hydrolysate |
| title_sort | metabolic engineering of i zymomonas mobilis i for xylonic acid production from lignocellulosic hydrolysate |
| topic | xylonic acid xylose dehydrogenase <i>Zymomonas mobilis</i> metabolic engineering lignocellulosic hydrolysate |
| url | https://www.mdpi.com/2311-5637/11/3/141 |
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