Details
| Original language | English |
|---|---|
| Article number | 57 |
| Journal | Microbial cell factories |
| Volume | 17 |
| Issue number | 1 |
| Early online date | 7 Apr 2018 |
| Publication status | Published - 2018 |
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.
Keywords
- Active inclusion bodies, AmbLOXe, Axolotl (Ambystoma mexicanum), In vitro wound healing assay, Lipoxygenase, Nanopills
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Microbial cell factories, Vol. 17, No. 1, 57, 2018.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl
AU - Stamm, Anne
AU - Straub, Sarah
AU - Vogt, Peter
AU - Scheper, Thomas
AU - Pepelanova, Iliyana
N1 - Funding: This work was performed within the framework of the BIOFABRICATION FOR NIFE initiative, financially supported by the Ministry of Economy and Culture (MWK) of Lower Saxony, Germany.
PY - 2018
Y1 - 2018
N2 - 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.
AB - 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.
KW - Active inclusion bodies
KW - AmbLOXe
KW - Axolotl (Ambystoma mexicanum)
KW - In vitro wound healing assay
KW - Lipoxygenase
KW - Nanopills
UR - http://www.scopus.com/inward/record.url?scp=85045065835&partnerID=8YFLogxK
U2 - 10.1186/s12934-018-0904-0
DO - 10.1186/s12934-018-0904-0
M3 - Article
C2 - 29626934
AN - SCOPUS:85045065835
VL - 17
JO - Microbial cell factories
JF - Microbial cell factories
SN - 1475-2859
IS - 1
M1 - 57
ER -