Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in Wines
Acetaldehyde is an important carbonyl compound commonly detected in wines. A high concentration of acetaldehyde can affect the flavor of wines and result in adverse effects on human health. Alcohol dehydrogenase I (ADH1) in <i>Saccharomyces cerevisiae</i> catalyzes the reduction reaction...
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MDPI AG
2023-12-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/12/1/38 |
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| author | Kun Geng Ying Lin Xueyun Zheng Cheng Li Shuting Chen He Ling Jun Yang Xiangyu Zhu Shuli Liang |
| author_facet | Kun Geng Ying Lin Xueyun Zheng Cheng Li Shuting Chen He Ling Jun Yang Xiangyu Zhu Shuli Liang |
| author_sort | Kun Geng |
| collection | DOAJ |
| description | Acetaldehyde is an important carbonyl compound commonly detected in wines. A high concentration of acetaldehyde can affect the flavor of wines and result in adverse effects on human health. Alcohol dehydrogenase I (ADH1) in <i>Saccharomyces cerevisiae</i> catalyzes the reduction reaction of acetaldehyde into ethanol in the presence of cofactors, showing the potential to reduce the content of acetaldehyde in wines. In this study, <i>ADH1</i> was successfully expressed in <i>Pichia pastoris</i> GS115 based on codon optimization. Then, the expression level of <i>ADH1</i> was enhanced by replacing its promoter with optimized promoters and increasing the copy number of the expression cassette, with ADH1 being purified using nickel column affinity chromatography. The enzymatic activity of purified ADH1 reached 605.44 ± 44.30 U/mg. The results of the effect of ADH1 on the content of acetaldehyde in wine revealed that the acetaldehyde content of wine samples was reduced from 168.05 ± 0.55 to 113.17 ± 6.08 mg/L with the addition of 5 mM NADH and the catalysis of ADH1, and from 135.53 ± 4.08 to 52.89 ± 2.20 mg/L through cofactor regeneration. Our study provides a novel approach to reducing the content of acetaldehyde in wines through enzymatic catalysis. |
| first_indexed | 2024-03-08T09:50:53Z |
| format | Article |
| id | doaj.art-704c2d5f3bd241f9be265f4501aa11c2 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-08T09:50:53Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-704c2d5f3bd241f9be265f4501aa11c22024-01-29T14:05:45ZengMDPI AGMicroorganisms2076-26072023-12-011213810.3390/microorganisms12010038Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in WinesKun Geng0Ying Lin1Xueyun Zheng2Cheng Li3Shuting Chen4He Ling5Jun Yang6Xiangyu Zhu7Shuli Liang8School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaDepartment of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USASchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaAcetaldehyde is an important carbonyl compound commonly detected in wines. A high concentration of acetaldehyde can affect the flavor of wines and result in adverse effects on human health. Alcohol dehydrogenase I (ADH1) in <i>Saccharomyces cerevisiae</i> catalyzes the reduction reaction of acetaldehyde into ethanol in the presence of cofactors, showing the potential to reduce the content of acetaldehyde in wines. In this study, <i>ADH1</i> was successfully expressed in <i>Pichia pastoris</i> GS115 based on codon optimization. Then, the expression level of <i>ADH1</i> was enhanced by replacing its promoter with optimized promoters and increasing the copy number of the expression cassette, with ADH1 being purified using nickel column affinity chromatography. The enzymatic activity of purified ADH1 reached 605.44 ± 44.30 U/mg. The results of the effect of ADH1 on the content of acetaldehyde in wine revealed that the acetaldehyde content of wine samples was reduced from 168.05 ± 0.55 to 113.17 ± 6.08 mg/L with the addition of 5 mM NADH and the catalysis of ADH1, and from 135.53 ± 4.08 to 52.89 ± 2.20 mg/L through cofactor regeneration. Our study provides a novel approach to reducing the content of acetaldehyde in wines through enzymatic catalysis.https://www.mdpi.com/2076-2607/12/1/38alcohol dehydrogenase I<i>Pichia pastoris</i>cofactor regenerationmulticopyacetaldehydewine |
| spellingShingle | Kun Geng Ying Lin Xueyun Zheng Cheng Li Shuting Chen He Ling Jun Yang Xiangyu Zhu Shuli Liang Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in Wines Microorganisms alcohol dehydrogenase I <i>Pichia pastoris</i> cofactor regeneration multicopy acetaldehyde wine |
| title | Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in Wines |
| title_full | Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in Wines |
| title_fullStr | Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in Wines |
| title_full_unstemmed | Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in Wines |
| title_short | Enhanced Expression of Alcohol Dehydrogenase I in <i>Pichia pastoris</i> Reduces the Content of Acetaldehyde in Wines |
| title_sort | enhanced expression of alcohol dehydrogenase i in i pichia pastoris i reduces the content of acetaldehyde in wines |
| topic | alcohol dehydrogenase I <i>Pichia pastoris</i> cofactor regeneration multicopy acetaldehyde wine |
| url | https://www.mdpi.com/2076-2607/12/1/38 |
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