Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl
Abstract Background AmbLOXe is a lipoxygenase, which is up-regulated during limb-redevelopment in the Mexican axolotl, Ambystoma mexicanum, an animal with remarkable regeneration capacity. Previous studies have shown that mammalian cells transformed with the gene of this epidermal lipoxygenase displ...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-04-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12934-018-0904-0 |
| _version_ | 1819153498076348416 |
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| author | Anne Stamm Sarah Strauß Peter Vogt Thomas Scheper Iliyana Pepelanova |
| author_facet | Anne Stamm Sarah Strauß Peter Vogt Thomas Scheper Iliyana Pepelanova |
| author_sort | Anne Stamm |
| collection | DOAJ |
| description | Abstract Background AmbLOXe is a lipoxygenase, which is up-regulated during limb-redevelopment in the Mexican axolotl, Ambystoma mexicanum, an animal with remarkable regeneration capacity. Previous studies have shown that mammalian cells transformed with the gene of this epidermal lipoxygenase display faster migration and wound closure rate during in vitro wound healing experiments. Results In this study, the gene of AmbLOXe was codon-optimized for expression in Escherichia coli and was produced in the insoluble fraction as protein aggregates. These inclusion bodies or nanopills were shown to be reservoirs containing functional protein during in vitro wound healing assays. For this purpose, functional inclusion bodies were used to coat cell culture surfaces prior cell seeding or were added directly to the medium after cells reached confluence. In both scenarios, AmbLOXe inclusion bodies led to faster migration rate and wound closure, in comparison to controls containing either no AmbLOXe or GFP inclusion bodies. Conclusions Our results demonstrate that AmbLOXe inclusion bodies are functional and may serve as stable reservoirs of this enzyme. Nevertheless, further studies with soluble enzyme are also necessary in order to start elucidating the exact molecular substrates of AmbLOXe and the biochemical pathways involved in the wound healing effect. |
| first_indexed | 2024-12-22T15:06:08Z |
| format | Article |
| id | doaj.art-aa63b2832c4f43b2b4157f480bad4b34 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-22T15:06:08Z |
| publishDate | 2018-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-aa63b2832c4f43b2b4157f480bad4b342022-12-21T18:21:58ZengBMCMicrobial Cell Factories1475-28592018-04-011711910.1186/s12934-018-0904-0Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotlAnne Stamm0Sarah Strauß1Peter Vogt2Thomas Scheper3Iliyana Pepelanova4Institute of Technical Chemistry, Gottfried Wilhelm Leibniz UniversityDepartment of Plastic, Aesthetic, Hand and Reconstructive Surgery, Hannover Medical SchoolDepartment of Plastic, Aesthetic, Hand and Reconstructive Surgery, Hannover Medical SchoolInstitute of Technical Chemistry, Gottfried Wilhelm Leibniz UniversityInstitute of Technical Chemistry, Gottfried Wilhelm Leibniz UniversityAbstract Background AmbLOXe is a lipoxygenase, which is up-regulated during limb-redevelopment in the Mexican axolotl, Ambystoma mexicanum, an animal with remarkable regeneration capacity. Previous studies have shown that mammalian cells transformed with the gene of this epidermal lipoxygenase display faster migration and wound closure rate during in vitro wound healing experiments. Results In this study, the gene of AmbLOXe was codon-optimized for expression in Escherichia coli and was produced in the insoluble fraction as protein aggregates. These inclusion bodies or nanopills were shown to be reservoirs containing functional protein during in vitro wound healing assays. For this purpose, functional inclusion bodies were used to coat cell culture surfaces prior cell seeding or were added directly to the medium after cells reached confluence. In both scenarios, AmbLOXe inclusion bodies led to faster migration rate and wound closure, in comparison to controls containing either no AmbLOXe or GFP inclusion bodies. Conclusions Our results demonstrate that AmbLOXe inclusion bodies are functional and may serve as stable reservoirs of this enzyme. Nevertheless, further studies with soluble enzyme are also necessary in order to start elucidating the exact molecular substrates of AmbLOXe and the biochemical pathways involved in the wound healing effect.http://link.springer.com/article/10.1186/s12934-018-0904-0AmbLOXeAxolotl (Ambystoma mexicanum)Active inclusion bodiesIn vitro wound healing assayLipoxygenaseNanopills |
| spellingShingle | Anne Stamm Sarah Strauß Peter Vogt Thomas Scheper Iliyana Pepelanova Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl Microbial Cell Factories AmbLOXe Axolotl (Ambystoma mexicanum) Active inclusion bodies In vitro wound healing assay Lipoxygenase Nanopills |
| title | Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl |
| title_full | Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl |
| title_fullStr | Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl |
| title_full_unstemmed | Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl |
| title_short | Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl |
| title_sort | positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the mexican axolotl |
| topic | AmbLOXe Axolotl (Ambystoma mexicanum) Active inclusion bodies In vitro wound healing assay Lipoxygenase Nanopills |
| url | http://link.springer.com/article/10.1186/s12934-018-0904-0 |
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