Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed Bioreactor
A recombinant <i>E. coli</i>, expressing nitrilase from <i>Acidovorax facilis</i> 72W with dual-site expression plasmid pRSFduet (<i>E. coli</i> pRSF-AfNit2), was constructed. It showed higher soluble expression of nitrilase than that in the pET21a plasmid. The re...
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MDPI AG
2023-02-01
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| Online Access: | https://www.mdpi.com/2073-4344/13/2/371 |
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| author | Xue-Jiao Liu Bao-Di Ma Xiao-Mei Wu Yi Xu |
| author_facet | Xue-Jiao Liu Bao-Di Ma Xiao-Mei Wu Yi Xu |
| author_sort | Xue-Jiao Liu |
| collection | DOAJ |
| description | A recombinant <i>E. coli</i>, expressing nitrilase from <i>Acidovorax facilis</i> 72W with dual-site expression plasmid pRSFduet (<i>E. coli</i> pRSF-AfNit2), was constructed. It showed higher soluble expression of nitrilase than that in the pET21a plasmid. The recombinant nitrilase can efficiently catalyze the hydrolysis of 3-cyanopyridine to nicotinic acid. The whole cells of <i>E. coli</i> pRSF-AfNit2 were immobilized by using sodium alginate/glutaraldehyde/polyethylene imine as the best immobilized reagents. The immobilized cells showed 95% activity recovery and excellent mechanical strength, with improved thermal stability and pH stability. They also retained 82% of initial activity after nearly two months of storage at 4 °C. A semi-continuous packed-bed bioreactor (sPBR) filled with the immobilized cells was studied for efficient production of nicotinic acid. After optimization, the highest space–time yield of 1576 g/(L·d) was obtained on 0.8 M substrate concentration at 2 mL/min of flow rate. The sPBR was repeatedly operated for 41 batches, keeping 100% conversion in the presence of 30 mM CaCl<sub>2</sub>. Finally, 95 g of nicotinic acid were obtained at 90% yield after separation and purification. The developed technology has potential application value. |
| first_indexed | 2024-03-11T09:00:37Z |
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| id | doaj.art-1bbf6f76c0fd49939657795e08b9be77 |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-11T09:00:37Z |
| publishDate | 2023-02-01 |
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| spelling | doaj.art-1bbf6f76c0fd49939657795e08b9be772023-11-16T19:42:12ZengMDPI AGCatalysts2073-43442023-02-0113237110.3390/catal13020371Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed BioreactorXue-Jiao Liu0Bao-Di Ma1Xiao-Mei Wu2Yi Xu3School of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418, ChinaSchool of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418, ChinaSchool of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418, ChinaSchool of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418, ChinaA recombinant <i>E. coli</i>, expressing nitrilase from <i>Acidovorax facilis</i> 72W with dual-site expression plasmid pRSFduet (<i>E. coli</i> pRSF-AfNit2), was constructed. It showed higher soluble expression of nitrilase than that in the pET21a plasmid. The recombinant nitrilase can efficiently catalyze the hydrolysis of 3-cyanopyridine to nicotinic acid. The whole cells of <i>E. coli</i> pRSF-AfNit2 were immobilized by using sodium alginate/glutaraldehyde/polyethylene imine as the best immobilized reagents. The immobilized cells showed 95% activity recovery and excellent mechanical strength, with improved thermal stability and pH stability. They also retained 82% of initial activity after nearly two months of storage at 4 °C. A semi-continuous packed-bed bioreactor (sPBR) filled with the immobilized cells was studied for efficient production of nicotinic acid. After optimization, the highest space–time yield of 1576 g/(L·d) was obtained on 0.8 M substrate concentration at 2 mL/min of flow rate. The sPBR was repeatedly operated for 41 batches, keeping 100% conversion in the presence of 30 mM CaCl<sub>2</sub>. Finally, 95 g of nicotinic acid were obtained at 90% yield after separation and purification. The developed technology has potential application value.https://www.mdpi.com/2073-4344/13/2/371nitrilasecell immobilizationnicotinic acidreaction optimizationsemi-continuous packed-bed bioreactor |
| spellingShingle | Xue-Jiao Liu Bao-Di Ma Xiao-Mei Wu Yi Xu Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed Bioreactor Catalysts nitrilase cell immobilization nicotinic acid reaction optimization semi-continuous packed-bed bioreactor |
| title | Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed Bioreactor |
| title_full | Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed Bioreactor |
| title_fullStr | Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed Bioreactor |
| title_full_unstemmed | Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed Bioreactor |
| title_short | Highly Efficient Biosynthesis of Nicotinic Acid by Immobilized Whole Cells of <i>E. coli</i> Expressing Nitrilase in Semi-Continuous Packed-Bed Bioreactor |
| title_sort | highly efficient biosynthesis of nicotinic acid by immobilized whole cells of i e coli i expressing nitrilase in semi continuous packed bed bioreactor |
| topic | nitrilase cell immobilization nicotinic acid reaction optimization semi-continuous packed-bed bioreactor |
| url | https://www.mdpi.com/2073-4344/13/2/371 |
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