Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i>
Myrosinases (EC 3.2.1.147) are enzymes known for the generation of hydrolysis products that have a potential beneficial effect on human health. Their reaction mechanisms are widely studied, in order to improve and optimize secondary metabolite production processes. In this work, kinetic and biochemi...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-01-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/12/2/233 |
| _version_ | 1797482257073569792 |
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| author | Carolina Curiqueo Andrea Mahn Antonio Castillo |
| author_facet | Carolina Curiqueo Andrea Mahn Antonio Castillo |
| author_sort | Carolina Curiqueo |
| collection | DOAJ |
| description | Myrosinases (EC 3.2.1.147) are enzymes known for the generation of hydrolysis products that have a potential beneficial effect on human health. Their reaction mechanisms are widely studied, in order to improve and optimize secondary metabolite production processes. In this work, kinetic and biochemical properties of the broccoli myrosinase enzyme produced from its cDNA cloned in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> were investigated. The results revealed that the thermal stability of the enzyme produced in <i>S. cerevisiae</i> was slightly higher (30 to 60 °C) than that of myrosinase produced in <i>E. coli</i> (20 to 50 °C). The effect of pH on the enzymatic activity was similar in both enzymes, with pH 3 being the optimum value under the reaction conditions used. The kinetic behavior of both enzymes was adjusted to the Michaelis–Menten model. The catalytic efficiency was up to 4 times higher in myrosinase produced in <i>S. cerevisiae</i>, compared to myrosinase produced in <i>E. coli</i>. The glycosylations present in the enzyme would be related to the formation of a dimeric quaternary structure and would not play an essential role in enzymatic activity, since both enzymes were biologically active. These results will probably allow the development of strategies for the production of bioactive metabolites of medical interest. |
| first_indexed | 2024-03-09T22:29:38Z |
| format | Article |
| id | doaj.art-2cfdf9a087af4966a1892c0ade92431f |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T22:29:38Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-2cfdf9a087af4966a1892c0ade92431f2023-11-23T18:58:37ZengMDPI AGBiomolecules2218-273X2022-01-0112223310.3390/biom12020233Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i>Carolina Curiqueo0Andrea Mahn1Antonio Castillo2Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile (USACH), Av. L. B. O’Higgins 3363, Estación Central, Santiago 9170022, ChileDepartamento de Ingeniería Química, Facultad de Ingeniería, Universidad de Santiago de Chile (USACH), Av. L. B. O’Higgins 3363, Estación Central, Santiago 9170022, ChileDepartamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile (USACH), Av. L. B. O’Higgins 3363, Estación Central, Santiago 9170022, ChileMyrosinases (EC 3.2.1.147) are enzymes known for the generation of hydrolysis products that have a potential beneficial effect on human health. Their reaction mechanisms are widely studied, in order to improve and optimize secondary metabolite production processes. In this work, kinetic and biochemical properties of the broccoli myrosinase enzyme produced from its cDNA cloned in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> were investigated. The results revealed that the thermal stability of the enzyme produced in <i>S. cerevisiae</i> was slightly higher (30 to 60 °C) than that of myrosinase produced in <i>E. coli</i> (20 to 50 °C). The effect of pH on the enzymatic activity was similar in both enzymes, with pH 3 being the optimum value under the reaction conditions used. The kinetic behavior of both enzymes was adjusted to the Michaelis–Menten model. The catalytic efficiency was up to 4 times higher in myrosinase produced in <i>S. cerevisiae</i>, compared to myrosinase produced in <i>E. coli</i>. The glycosylations present in the enzyme would be related to the formation of a dimeric quaternary structure and would not play an essential role in enzymatic activity, since both enzymes were biologically active. These results will probably allow the development of strategies for the production of bioactive metabolites of medical interest.https://www.mdpi.com/2218-273X/12/2/233broccoli myrosinasecDNA expression<i>Escherichia coli</i><i>Saccharomyces cerevisiae</i> |
| spellingShingle | Carolina Curiqueo Andrea Mahn Antonio Castillo Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> Biomolecules broccoli myrosinase cDNA expression <i>Escherichia coli</i> <i>Saccharomyces cerevisiae</i> |
| title | Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> |
| title_full | Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> |
| title_fullStr | Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> |
| title_full_unstemmed | Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> |
| title_short | Broccoli Myrosinase cDNA Expression in <i>Escherichia coli</i> and <i>Saccharomyces cerevisiae</i> |
| title_sort | broccoli myrosinase cdna expression in i escherichia coli i and i saccharomyces cerevisiae i |
| topic | broccoli myrosinase cDNA expression <i>Escherichia coli</i> <i>Saccharomyces cerevisiae</i> |
| url | https://www.mdpi.com/2218-273X/12/2/233 |
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