Engineering Patatin for enhanced lipase activity and long-chain fatty acid specificity via rational design.
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| Autores principales: | , , , , , , , , , , |
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| Formato: | Artículo científico |
| Lenguaje: | en |
| Publicado: |
Food chemistry
2025
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| _version_ | 1868266216514125826 |
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| author | Nie, Ting Bai, Xue Liao, Zikang Chen, Runsha Le, Qianyu Zhang, Yong Liu, Xiaodan Bian, Xiaoying Wu, Shuhong Wu, Jinhong Li, Xianghong |
| author_facet | Nie, Ting Bai, Xue Liao, Zikang Chen, Runsha Le, Qianyu Zhang, Yong Liu, Xiaodan Bian, Xiaoying Wu, Shuhong Wu, Jinhong Li, Xianghong Nie, Ting Bai, Xue Liao, Zikang Chen, Runsha Le, Qianyu Zhang, Yong Liu, Xiaodan Bian, Xiaoying Wu, Shuhong Wu, Jinhong Li, Xianghong |
| collection | PubMed - marine biology |
| contents | Engineering Patatin for enhanced lipase activity and long-chain fatty acid specificity via rational design. Nie, Ting Bai, Xue Liao, Zikang Chen, Runsha Le, Qianyu Zhang, Yong Liu, Xiaodan Bian, Xiaoying Wu, Shuhong Wu, Jinhong Li, Xianghong Lipase Substrate Specificity Protein Engineering Fatty Acids Plant Proteins Enzyme Stability Saccharomycetales Kinetics Biocatalysis Patatin, a multifunctional glycoprotein from potatoes, is a promising lipase for industrial applications due to its emulsifying, antioxidant, and lipid-modifying properties. However, its low expression efficiency and preference for short-chain substrates limit practical utility. Here, we addressed these challenges by heterologously expressing patatin in Pichia pastoris X-33, achieving a yield of 121 mg/L through optimized fermentation. The enzyme showed optimal activity at 35 °C and pH 10.0, with methanol enhancing activity, while Fe/Fe inhibited it. Rational design of the D286A mutant significantly improved long-chain substrate specificity (3.2-fold higher activity for pNP-C16) and thermal stability (ΔTm = +5.4 °C). Molecular dynamics revealed that the mutation disrupted an α-helix (residues 280-286), forming a flexible loop to accommodate long-chain substrates via hydrogen bonding and π-alkyl interactions. Structural integrity was confirmed by circular dichroism. This work provides a scalable platform for engineering patatin, with future studies targeting industrial-scale production and applications in functional lipids and biocatalysis. |
| format | Artículo científico |
| id | pubmed_40203696 |
| institution | PubMed |
| language | en |
| publishDate | 2025 |
| publisher | Food chemistry |
| record_format | pubmed |
| spellingShingle | Engineering Patatin for enhanced lipase activity and long-chain fatty acid specificity via rational design. Nie, Ting Bai, Xue Liao, Zikang Chen, Runsha Le, Qianyu Zhang, Yong Liu, Xiaodan Bian, Xiaoying Wu, Shuhong Wu, Jinhong Li, Xianghong Lipase Substrate Specificity Protein Engineering Fatty Acids Plant Proteins Enzyme Stability Saccharomycetales Kinetics Biocatalysis Engineering Patatin for enhanced lipase activity and long-chain fatty acid specificity via rational design. Nie, Ting Bai, Xue Liao, Zikang Chen, Runsha Le, Qianyu Zhang, Yong Liu, Xiaodan Bian, Xiaoying Wu, Shuhong Wu, Jinhong Li, Xianghong Lipase Substrate Specificity Protein Engineering Fatty Acids Plant Proteins Enzyme Stability Saccharomycetales Kinetics Biocatalysis Patatin, a multifunctional glycoprotein from potatoes, is a promising lipase for industrial applications due to its emulsifying, antioxidant, and lipid-modifying properties. However, its low expression efficiency and preference for short-chain substrates limit practical utility. Here, we addressed these challenges by heterologously expressing patatin in Pichia pastoris X-33, achieving a yield of 121 mg/L through optimized fermentation. The enzyme showed optimal activity at 35 °C and pH 10.0, with methanol enhancing activity, while Fe/Fe inhibited it. Rational design of the D286A mutant significantly improved long-chain substrate specificity (3.2-fold higher activity for pNP-C16) and thermal stability (ΔTm = +5.4 °C). Molecular dynamics revealed that the mutation disrupted an α-helix (residues 280-286), forming a flexible loop to accommodate long-chain substrates via hydrogen bonding and π-alkyl interactions. Structural integrity was confirmed by circular dichroism. This work provides a scalable platform for engineering patatin, with future studies targeting industrial-scale production and applications in functional lipids and biocatalysis. |
| title | Engineering Patatin for enhanced lipase activity and long-chain fatty acid specificity via rational design. |
| topic | Lipase Substrate Specificity Protein Engineering Fatty Acids Plant Proteins Enzyme Stability Saccharomycetales Kinetics Biocatalysis |
| url | https://pubmed.ncbi.nlm.nih.gov/40203696/ |