Journal
ADVANCED SCIENCE
Volume 9, Issue 7, Pages -Publisher
WILEY
DOI: 10.1002/advs.202103503
Keywords
artificial melanosome; o-nitrobenzyl tyrosine; photoactivatable tyrosinase; skin protection; transdermal delivery
Categories
Funding
- National Research Foundation of Korea (NRF) - Ministry of Science, ICT and Future Planning [NRF-2021R1A2C2008821]
- Korea Medical Device Development Fund grant - Korea government (Ministry of Science and ICT) [KMDF_PR_20200901_0151]
- Korea Medical Device Development Fund grant - Korea government (Ministry of Trade, Industry and Energy) [KMDF_PR_20200901_0151]
- Korea Medical Device Development Fund grant - Korea government (Ministry of Health and Welfare) [KMDF_PR_20200901_0151]
- Korea Medical Device Development Fund grant - Korea government (Ministry of Food and Drug Safety) [KMDF_PR_20200901_0151]
- 'R&D Program for Forest Science Technology by Korea Forest Service (Korea Forestry Promotion Institute) [2021405B10-2123-0101]
- Dong-A University research fund
- Korea Forestry Promotion Institute (KOFPI) [2021405B10-2123-0101] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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This study presents a method for in situ biosynthesis of artificial melanin in skin using photoactivatable tyrosinase. By introducing a photocleavable o-nitrobenzyl tyrosine into the tyrosinase, its enzymatic activity can be recovered through photo-cleavage. Using this method, melanin particles and hydrogel can be synthesized in the skin for tissue protection. Encapsulating the photoactivatable tyrosinase into lipid nanoparticles allows transdermal delivery into the skin tissue.
Tyrosinase-mediated melanin synthesis is an essential biological process that can protect skin from UV radiation and radical species. This work reports on in situ biosynthesis of artificial melanin in native skin using photoactivatable tyrosinase (PaTy). The I41Y mutant of Streptomyces avermitilis tyrosinase (SaTy) shows enzymatic activity comparable to that of wild-type SaTy. This Y41 is replaced with photocleavable o-nitrobenzyl tyrosine (ONBY) using the introduction of amber codon and ONBY-tRNA synthetase/tRNA pairs. The ONBY efficiently blocks the active site and tyrosinase activity is rapidly recovered by the photo-cleavage of ONBY. The activated PaTy successfully oxidizes L-tyrosine and tyramine-conjugated hyaluronic acid (HA_T) to synthesize melanin particles and hydrogel, respectively. To produce artificial melanin in living tissues, PaTy is encapsulated into lipid nanoparticles as an artificial melanosome. Using liposomes containing PaTy (PaTy_Lip), PaTy is transdermally delivered into ex vivo porcine skin and in vivo mouse skin tissues, thus achieving the in situ biosynthesis of artificial melanin for skin tissue protection under UV irradiation. The results of this study demonstrate that this biomimetic system can recapitulate the biosynthetic analogs of naturally occurring melanin. It should therefore be considered to be a promising strategy for producing protective biological molecules within living systems for tissue protection.
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