A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142
Phosphite dehydrogenase (PtxD) is a promising enzyme for NAD(P)H regeneration. To expand the usability of PtxD, we cloned, expressed, and analyzed PtxD from the marine cyanobacterium Cyanothece sp. ATCC 51142 (Ct-PtxD). Ct-PtxD exhibited maximum activity at pH 9.0°C and 50°C and high stability over...
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| Format: | Article |
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Frontiers Media S.A.
2023-08-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1255582/full |
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| author | Gamal Nasser Abdel-Hady Gamal Nasser Abdel-Hady Takahisa Tajima Takahisa Tajima Takeshi Ikeda Takenori Ishida Hisakage Funabashi Hisakage Funabashi Akio Kuroda Akio Kuroda Ryuichi Hirota Ryuichi Hirota |
| author_facet | Gamal Nasser Abdel-Hady Gamal Nasser Abdel-Hady Takahisa Tajima Takahisa Tajima Takeshi Ikeda Takenori Ishida Hisakage Funabashi Hisakage Funabashi Akio Kuroda Akio Kuroda Ryuichi Hirota Ryuichi Hirota |
| author_sort | Gamal Nasser Abdel-Hady |
| collection | DOAJ |
| description | Phosphite dehydrogenase (PtxD) is a promising enzyme for NAD(P)H regeneration. To expand the usability of PtxD, we cloned, expressed, and analyzed PtxD from the marine cyanobacterium Cyanothece sp. ATCC 51142 (Ct-PtxD). Ct-PtxD exhibited maximum activity at pH 9.0°C and 50°C and high stability over a wide pH range of 6.0–10.0. Compared to previously reported PtxDs, Ct-PtxD showed increased resistance to salt ions such as Na+, K+, and NH4+. It also exhibited high tolerance to organic solvents such as ethanol, dimethylformamide, and methanol when bound to its preferred cofactor, NAD+. Remarkably, these organic solvents enhanced the Ct-PtxD activity while inhibiting the PtxD activity of Ralstonia sp. 4506 (Rs-PtxD) at concentrations ranging from 10% to 30%. Molecular electrostatic potential analysis showed that the NAD+-binding site of Ct-PtxD was rich in positively charged residues, which may attract the negatively charged pyrophosphate group of NAD+ under high-salt conditions. Amino acid composition analysis revealed that Ct-PtxD contained fewer hydrophobic amino acids than other PtxD enzymes, which reduced the hydrophobicity and increased the hydration of protein surface under low water activity. We also demonstrated that the NADH regeneration system using Ct-PtxD is useful for the coupled chiral conversion of trimethylpyruvic acid into L-tert-leucine using leucine dehydrogenase under high ammonium conditions, which is less supported by the Rs-PtxD enzyme. These results imply that Ct-PtxD might be a potential candidate for NAD(P)H regeneration in industrial applications under the reaction conditions containing salt and organic solvent. |
| first_indexed | 2024-03-12T14:22:11Z |
| format | Article |
| id | doaj.art-36d2e4223af349a7b6cd7fd114148614 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-03-12T14:22:11Z |
| publishDate | 2023-08-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-36d2e4223af349a7b6cd7fd1141486142023-08-18T16:14:13ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-08-011110.3389/fbioe.2023.12555821255582A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142Gamal Nasser Abdel-Hady0Gamal Nasser Abdel-Hady1Takahisa Tajima2Takahisa Tajima3Takeshi Ikeda4Takenori Ishida5Hisakage Funabashi6Hisakage Funabashi7Akio Kuroda8Akio Kuroda9Ryuichi Hirota10Ryuichi Hirota11Unit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanDepartment of Genetics, Faculty of Agriculture, Minia University, Minia, EgyptUnit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanSeto Inland Sea Carbon-neutral Research Center, Hiroshima University, Hiroshima, JapanUnit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanUnit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanUnit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanSeto Inland Sea Carbon-neutral Research Center, Hiroshima University, Hiroshima, JapanUnit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanSeto Inland Sea Carbon-neutral Research Center, Hiroshima University, Hiroshima, JapanUnit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanSeto Inland Sea Carbon-neutral Research Center, Hiroshima University, Hiroshima, JapanPhosphite dehydrogenase (PtxD) is a promising enzyme for NAD(P)H regeneration. To expand the usability of PtxD, we cloned, expressed, and analyzed PtxD from the marine cyanobacterium Cyanothece sp. ATCC 51142 (Ct-PtxD). Ct-PtxD exhibited maximum activity at pH 9.0°C and 50°C and high stability over a wide pH range of 6.0–10.0. Compared to previously reported PtxDs, Ct-PtxD showed increased resistance to salt ions such as Na+, K+, and NH4+. It also exhibited high tolerance to organic solvents such as ethanol, dimethylformamide, and methanol when bound to its preferred cofactor, NAD+. Remarkably, these organic solvents enhanced the Ct-PtxD activity while inhibiting the PtxD activity of Ralstonia sp. 4506 (Rs-PtxD) at concentrations ranging from 10% to 30%. Molecular electrostatic potential analysis showed that the NAD+-binding site of Ct-PtxD was rich in positively charged residues, which may attract the negatively charged pyrophosphate group of NAD+ under high-salt conditions. Amino acid composition analysis revealed that Ct-PtxD contained fewer hydrophobic amino acids than other PtxD enzymes, which reduced the hydrophobicity and increased the hydration of protein surface under low water activity. We also demonstrated that the NADH regeneration system using Ct-PtxD is useful for the coupled chiral conversion of trimethylpyruvic acid into L-tert-leucine using leucine dehydrogenase under high ammonium conditions, which is less supported by the Rs-PtxD enzyme. These results imply that Ct-PtxD might be a potential candidate for NAD(P)H regeneration in industrial applications under the reaction conditions containing salt and organic solvent.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1255582/fullphosphite dehydrogenaseNAD(P)H regenerationsalt resistanceorganic solvent tolerancelow water activity |
| spellingShingle | Gamal Nasser Abdel-Hady Gamal Nasser Abdel-Hady Takahisa Tajima Takahisa Tajima Takeshi Ikeda Takenori Ishida Hisakage Funabashi Hisakage Funabashi Akio Kuroda Akio Kuroda Ryuichi Hirota Ryuichi Hirota A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142 Frontiers in Bioengineering and Biotechnology phosphite dehydrogenase NAD(P)H regeneration salt resistance organic solvent tolerance low water activity |
| title | A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142 |
| title_full | A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142 |
| title_fullStr | A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142 |
| title_full_unstemmed | A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142 |
| title_short | A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142 |
| title_sort | novel salt and organic solvent tolerant phosphite dehydrogenase from cyanothece sp atcc 51142 |
| topic | phosphite dehydrogenase NAD(P)H regeneration salt resistance organic solvent tolerance low water activity |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1255582/full |
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