Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i>
Tyrosine aminotransferase (TAT, E.C. 2.6.1.5) is a pyridoxal phosphate-dependent aminotransferase that is widely found in living organisms. It catalyzes the transfer of the amino group on tyrosine to α-ketoglutarate to produce 4-hydroxyphenylpyruvic acid (4-HPP) and is the first enzyme for tyrosine...
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MDPI AG
2023-10-01
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| Series: | International Journal of Molecular Sciences |
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| author | Shuai Dong Long Wang Huiting Qin Hongbin Zhan Donghao Wang Xiaoyan Cao |
| author_facet | Shuai Dong Long Wang Huiting Qin Hongbin Zhan Donghao Wang Xiaoyan Cao |
| author_sort | Shuai Dong |
| collection | DOAJ |
| description | Tyrosine aminotransferase (TAT, E.C. 2.6.1.5) is a pyridoxal phosphate-dependent aminotransferase that is widely found in living organisms. It catalyzes the transfer of the amino group on tyrosine to α-ketoglutarate to produce 4-hydroxyphenylpyruvic acid (4-HPP) and is the first enzyme for tyrosine degradation. Three <i>SmTATs</i> have been identified in the genome of <i>Salvia miltiorrhiza</i> (a model medicinal plant), but their information is very limited. Here, the expression profiles of the three <i>SmTAT</i> genes (<i>SmTAT1</i>, <i>SmTAT2</i>, and <i>SmTAT3</i>) were studied. All three genes expressed in different tissues and responded to methyl jasmonate stimuli. <i>Sm</i>TAT proteins are localized in the cytoplasm. The recombinant <i>Sm</i>TATs were subjected to in vitro biochemical properties. All three recombinant enzymes had TAT activities and <i>Sm</i>TAT1 had the highest catalytic activity for tyrosine, followed by <i>Sm</i>TAT3. Also, <i>Sm</i>TAT1 preferred the direction of tyrosine deamination to 4-HPP, while <i>Sm</i>TAT2 preferred transamination of 4-HPP to tyrosine. In parallel, transient overexpression of <i>SmTATs</i> in tobacco leaves revealed that all three <i>Sm</i>TAT proteins catalyzed tyrosine to 4-HPP in vivo, with <i>Sm</i>TAT1 exhibiting the highest enzymatic activity. Overall, our results lay a foundation for the production of tyrosine-derived secondary metabolites via metabolic engineering or synthetic biology in the future. |
| first_indexed | 2024-03-11T11:29:12Z |
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| language | English |
| last_indexed | 2024-03-11T11:29:12Z |
| publishDate | 2023-10-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e14cf9f927c7465992bd406f4449fffa2023-11-10T15:04:29ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-10-0124211557510.3390/ijms242115575Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i>Shuai Dong0Long Wang1Huiting Qin2Hongbin Zhan3Donghao Wang4Xiaoyan Cao5Key Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, College of Life Science, Shaanxi Normal University, Xi’an 710062, ChinaKey Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, College of Life Science, Shaanxi Normal University, Xi’an 710062, ChinaKey Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, College of Life Science, Shaanxi Normal University, Xi’an 710062, ChinaKey Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, College of Life Science, Shaanxi Normal University, Xi’an 710062, ChinaKey Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, College of Life Science, Shaanxi Normal University, Xi’an 710062, ChinaKey Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, College of Life Science, Shaanxi Normal University, Xi’an 710062, ChinaTyrosine aminotransferase (TAT, E.C. 2.6.1.5) is a pyridoxal phosphate-dependent aminotransferase that is widely found in living organisms. It catalyzes the transfer of the amino group on tyrosine to α-ketoglutarate to produce 4-hydroxyphenylpyruvic acid (4-HPP) and is the first enzyme for tyrosine degradation. Three <i>SmTATs</i> have been identified in the genome of <i>Salvia miltiorrhiza</i> (a model medicinal plant), but their information is very limited. Here, the expression profiles of the three <i>SmTAT</i> genes (<i>SmTAT1</i>, <i>SmTAT2</i>, and <i>SmTAT3</i>) were studied. All three genes expressed in different tissues and responded to methyl jasmonate stimuli. <i>Sm</i>TAT proteins are localized in the cytoplasm. The recombinant <i>Sm</i>TATs were subjected to in vitro biochemical properties. All three recombinant enzymes had TAT activities and <i>Sm</i>TAT1 had the highest catalytic activity for tyrosine, followed by <i>Sm</i>TAT3. Also, <i>Sm</i>TAT1 preferred the direction of tyrosine deamination to 4-HPP, while <i>Sm</i>TAT2 preferred transamination of 4-HPP to tyrosine. In parallel, transient overexpression of <i>SmTATs</i> in tobacco leaves revealed that all three <i>Sm</i>TAT proteins catalyzed tyrosine to 4-HPP in vivo, with <i>Sm</i>TAT1 exhibiting the highest enzymatic activity. Overall, our results lay a foundation for the production of tyrosine-derived secondary metabolites via metabolic engineering or synthetic biology in the future.https://www.mdpi.com/1422-0067/24/21/15575tyrosine aminotransferase<i>Salvia miltiorrhiza</i>catalytic activitytransient over-expression |
| spellingShingle | Shuai Dong Long Wang Huiting Qin Hongbin Zhan Donghao Wang Xiaoyan Cao Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i> International Journal of Molecular Sciences tyrosine aminotransferase <i>Salvia miltiorrhiza</i> catalytic activity transient over-expression |
| title | Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i> |
| title_full | Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i> |
| title_fullStr | Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i> |
| title_full_unstemmed | Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i> |
| title_short | Expression Patterns and Functional Analysis of Three <i>SmTAT</i> Genes Encoding Tyrosine Aminotransferases in <i>Salvia miltiorrhiza</i> |
| title_sort | expression patterns and functional analysis of three i smtat i genes encoding tyrosine aminotransferases in i salvia miltiorrhiza i |
| topic | tyrosine aminotransferase <i>Salvia miltiorrhiza</i> catalytic activity transient over-expression |
| url | https://www.mdpi.com/1422-0067/24/21/15575 |
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