Truncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide
Abstract Formate dehydrogenase (FDH) is widely applied in regeneration of redox cofactors. There are continuing interests to engineer FDH for improved catalytic activity and cofactor preference. In the crystal structure of FDH from Pseudomonas sp. 101 (pseFDH), the C terminus with 9 amino acid resid...
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Nature Portfolio
2024-11-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-79885-z |
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| author | Xiaojia Guo Xueying Wang Yinghan Hu Lingyun Zhang Zongbao K. Zhao |
| author_facet | Xiaojia Guo Xueying Wang Yinghan Hu Lingyun Zhang Zongbao K. Zhao |
| author_sort | Xiaojia Guo |
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| description | Abstract Formate dehydrogenase (FDH) is widely applied in regeneration of redox cofactors. There are continuing interests to engineer FDH for improved catalytic activity and cofactor preference. In the crystal structure of FDH from Pseudomonas sp. 101 (pseFDH), the C terminus with 9 amino acid residues cannot be resolved. However, our earlier work showed mutations at C terminus led pseFDH variants to favor a non-natural cofactor nicotinamide cytosine dinucleotide (NCD). Here, we investigated the role of C-terminal residues on cofactor preference by truncating their corresponding C terminus of pseFDH variants. Sequence comparison analysis showed that C-terminal residues were barely conservative among different FDHs. pseFDH and mutants with their C termini truncated were constructed, and the resulted variants showed improved preference to NCD mainly because NAD-dependent activity dropped more substantially. Further structure analysis showed that these pseFDH variants had their cofactor binding domains reconstructed to favor molecular interactions with NCD. Our work indicated that C-terminal residues of pseFDH affected enzyme activity and cofactor preference, which provides a new approach for ameliorating the performance of redox enzymes. |
| format | Article |
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| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
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| spelling | doaj-art-eaf2b6db501a4ec49b3424656facff512025-08-20T02:22:16ZengNature PortfolioScientific Reports2045-23222024-11-011411810.1038/s41598-024-79885-zTruncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotideXiaojia Guo0Xueying Wang1Yinghan Hu2Lingyun Zhang3Zongbao K. Zhao4MOE Key Laboratory of Bio-Intelligent Manufacturing, School of Bioengineering, Dalian University of TechnologyLaboratory of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of SciencesLaboratory of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of SciencesLaboratory of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of SciencesMOE Key Laboratory of Bio-Intelligent Manufacturing, School of Bioengineering, Dalian University of TechnologyAbstract Formate dehydrogenase (FDH) is widely applied in regeneration of redox cofactors. There are continuing interests to engineer FDH for improved catalytic activity and cofactor preference. In the crystal structure of FDH from Pseudomonas sp. 101 (pseFDH), the C terminus with 9 amino acid residues cannot be resolved. However, our earlier work showed mutations at C terminus led pseFDH variants to favor a non-natural cofactor nicotinamide cytosine dinucleotide (NCD). Here, we investigated the role of C-terminal residues on cofactor preference by truncating their corresponding C terminus of pseFDH variants. Sequence comparison analysis showed that C-terminal residues were barely conservative among different FDHs. pseFDH and mutants with their C termini truncated were constructed, and the resulted variants showed improved preference to NCD mainly because NAD-dependent activity dropped more substantially. Further structure analysis showed that these pseFDH variants had their cofactor binding domains reconstructed to favor molecular interactions with NCD. Our work indicated that C-terminal residues of pseFDH affected enzyme activity and cofactor preference, which provides a new approach for ameliorating the performance of redox enzymes.https://doi.org/10.1038/s41598-024-79885-zFormate dehydrogenaseDirected evolutionCofactor preferenceC terminusNon-natural cofactor |
| spellingShingle | Xiaojia Guo Xueying Wang Yinghan Hu Lingyun Zhang Zongbao K. Zhao Truncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide Scientific Reports Formate dehydrogenase Directed evolution Cofactor preference C terminus Non-natural cofactor |
| title | Truncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide |
| title_full | Truncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide |
| title_fullStr | Truncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide |
| title_full_unstemmed | Truncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide |
| title_short | Truncating the C terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide |
| title_sort | truncating the c terminus of formate dehydrogenase leads to improved preference to nicotinamide cytosine dinucleotide |
| topic | Formate dehydrogenase Directed evolution Cofactor preference C terminus Non-natural cofactor |
| url | https://doi.org/10.1038/s41598-024-79885-z |
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