Two possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluation
Epiphytic bacteria from marine macroalgae synthesize enzymes of industrial and biotechnological interest. In this study, we obtained two DNA candidate fragments for lipid-degrading enzymes from the total DNA of Ulva lactuca-associated epiphytic bacteria. First, we evaluated a method for total ba...
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| Format: | Article |
| Language: | English |
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Pontificia Universidad Javeriana
2020-07-01
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| Series: | Universitas Scientiarum |
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| Online Access: | https://revistas.javeriana.edu.co/index.php/scientarium/article/view/25583 |
| _version_ | 1818843803198423040 |
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| author | Ruiz-Toquica Jordan Steven, Comba-González Natalia Beatriz, Montoya-Castaño Dolly |
| author_facet | Ruiz-Toquica Jordan Steven, Comba-González Natalia Beatriz, Montoya-Castaño Dolly |
| author_sort | Ruiz-Toquica Jordan Steven, Comba-González Natalia Beatriz, Montoya-Castaño Dolly |
| collection | DOAJ |
| description | Epiphytic bacteria from marine macroalgae synthesize enzymes of
industrial and biotechnological interest. In this study, we obtained two
DNA candidate fragments for lipid-degrading enzymes from the total
DNA of Ulva lactuca-associated epiphytic bacteria. First, we evaluated a
method for total bacterial DNA isolation from the surface of U. lactuca
thalli. Then, we designed sets of primers and used them directly for
PCR amplification. The resulting PCR products were sequence-analyzed
and used for expression and functional evaluation with the Escherichia
coli pBAD-TOPO system. We obtained high molecular weight and
good quality total bacterial DNA that served as a template to identify
a fragment corresponding to an Acetyl-CoA C-Acetyltransferase (or
Thiolase), and a candidate fragment for a versatile “true” lipase. We
expressed the possible “true” lipase gene fragment heterologously in
Escherichia coli and obtained proof of hydrolytic activity on Tributyrin,
Tween-20, and Olive-oil media. This study resulted in new knowledge
on U. lactuca-associated epiphytic bacteria as possible brand-new sources
of enzymes such as thiolases and “true” lipases. However, future studies
are required to describe the characteristics and important applications of
these candidate enzymes. |
| first_indexed | 2024-12-19T05:03:40Z |
| format | Article |
| id | doaj.art-1b4b40d9736a4faca8c63bb122e99d55 |
| institution | Directory Open Access Journal |
| issn | 0122-7483 2027-1352 |
| language | English |
| last_indexed | 2024-12-19T05:03:40Z |
| publishDate | 2020-07-01 |
| publisher | Pontificia Universidad Javeriana |
| record_format | Article |
| series | Universitas Scientiarum |
| spelling | doaj.art-1b4b40d9736a4faca8c63bb122e99d552022-12-21T20:35:01ZengPontificia Universidad JaverianaUniversitas Scientiarum0122-74832027-13522020-07-0125224727510.11144/Javeriana.SC25-2.tpceTwo possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluationRuiz-Toquica Jordan Steven, Comba-González Natalia Beatriz, Montoya-Castaño Dolly0Grupo de Bioprocesos y Bioprospección, Instituto de Biotecnología, Universidad Nacional de Colombia, Carrera 45 No. 26 – 85, Bogotá, Colombia, ZIP Code: 111321.Epiphytic bacteria from marine macroalgae synthesize enzymes of industrial and biotechnological interest. In this study, we obtained two DNA candidate fragments for lipid-degrading enzymes from the total DNA of Ulva lactuca-associated epiphytic bacteria. First, we evaluated a method for total bacterial DNA isolation from the surface of U. lactuca thalli. Then, we designed sets of primers and used them directly for PCR amplification. The resulting PCR products were sequence-analyzed and used for expression and functional evaluation with the Escherichia coli pBAD-TOPO system. We obtained high molecular weight and good quality total bacterial DNA that served as a template to identify a fragment corresponding to an Acetyl-CoA C-Acetyltransferase (or Thiolase), and a candidate fragment for a versatile “true” lipase. We expressed the possible “true” lipase gene fragment heterologously in Escherichia coli and obtained proof of hydrolytic activity on Tributyrin, Tween-20, and Olive-oil media. This study resulted in new knowledge on U. lactuca-associated epiphytic bacteria as possible brand-new sources of enzymes such as thiolases and “true” lipases. However, future studies are required to describe the characteristics and important applications of these candidate enzymes.https://revistas.javeriana.edu.co/index.php/scientarium/article/view/25583epiphytic bacteria; lipases; pcr; thiolases; ulva lactuca. |
| spellingShingle | Ruiz-Toquica Jordan Steven, Comba-González Natalia Beatriz, Montoya-Castaño Dolly Two possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluation Universitas Scientiarum epiphytic bacteria; lipases; pcr; thiolases; ulva lactuca. |
| title | Two possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluation |
| title_full | Two possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluation |
| title_fullStr | Two possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluation |
| title_full_unstemmed | Two possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluation |
| title_short | Two possible candidate enzymes from Ulva lactuca-associated epiphytic bacteria obtained through PCR and functional evaluation |
| title_sort | two possible candidate enzymes from ulva lactuca associated epiphytic bacteria obtained through pcr and functional evaluation |
| topic | epiphytic bacteria; lipases; pcr; thiolases; ulva lactuca. |
| url | https://revistas.javeriana.edu.co/index.php/scientarium/article/view/25583 |
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