A highly efficient and cost-effective recombinant production of a bacterial photolyase from the Antarctic isolate Hymenobacter sp. UV11
Photolyases are DNA-repairing flavoproteins that are represented in most phylogenetic taxa with the exception of placental mammals. These enzymes reduce the ultraviolet-induced DNA damage; thus, they have features that make them very attractive for dermatological or other medical uses, such as the prevention of human skin cancer and actinic keratosis. In this work, we identified a 50.8 kDa photolyase from the UVC-resistant Antarctic bacterium Hymenobacter sp. UV11. The enzyme was produced by recombinant DNA technology, purified using immobilized metal affinity chromatography and its activity was analyzed using different approaches: detection of cyclobutane pyrimidine dimers (CPDs) by immunochemistry, high-performance liquid chromatography and comet assays using Chinese Hamster Ovary (CHO) and immortalized nontumorigenic human epidermal (HaCat) cells. The information supports that the recombinant protein has the ability to repair the formation of CPDs, on both double- and single-stranded DNA. This CPD-photolyase was fully active on CHO and HaCat cell lines, suggesting that this enzyme could be used for medical or cosmetic purposes. Results also suggest that the UV11 CPD-photolyase uses MTHF as chromophore in the antenna domain. The potential use of this recombinant enzyme in the development of new inventions with pharmaceutical and cosmetic applications is discussed during this work.
KeywordsPhotolyase Hymenobacter UV-irradiation Photorepair Antarctica
The authors thank the Uruguayan Antarctic Institute for the logistic support during the stay in the Antarctic Base Artigas. S. Castro-Sowinski, W. Martinez Lopez and J. J. Marizcurrena are members of the National Research System (SNI, Sistema Nacional de Investigadores). This work was partially supported by PEDECIBA (Programa de Desarrollo de las Ciencias Básicas), CSIC (Comisión Sectorial de Investigación Científica; Project C667), ANII (Agencia Nacional de Investigación e Innovación, Project FMV_3_2016_1_1226654) and donations by Celsius Laboratory (http://www.celsius.uy/). The work of JJM was supported by ANII and CAP (Comisión Académica de Posgrado, UdelaR).
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Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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