Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl

Microbial Cell Factories

Sarah Strauß¹  Peter Vogt¹  Iliyana Pepelanova²  Anne Stamm²  Thomas Scheper² 

[1] Department of Plastic, Aesthetic, Hand and Reconstructive Surgery, Hannover Medical School;Institute of Technical Chemistry, Gottfried Wilhelm Leibniz University;

关键词: AmbLOXe;    Axolotl (Ambystoma mexicanum);    Active inclusion bodies;    In vitro wound healing assay;    Lipoxygenase;    Nanopills;   

DOI  :  10.1186/s12934-018-0904-0

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown