Chemical Characterization and Biotechnological Applicability of Pigments Isolated from Antarctic Bacteria
Considering the global trend in the search for alternative natural compounds with antioxidant and sun protection factor (SPF) boosting properties, bacterial carotenoids represent an opportunity for exploring pigments of natural origin which possess high antioxidant activity, lower toxicity, no residues, and no environmental risk and are readily decomposable. In this work, three pigmented bacteria from the Antarctic continent, named Arthrobacter agilis 50cyt, Zobellia laminarie 465, and Arthrobacter psychrochitiniphilus 366, were able to withstand UV-B and UV-C radiation. The pigments were extracted and tested for UV absorption, antioxidant capacity, photostability, and phototoxicity profile in murine fibroblasts (3T3 NRU PT–OECD TG 432) to evaluate their further potential use as UV filters. Furthermore, the pigments were identified by ultra-high-performance liquid chromatography–photodiode array detector–mass spectrometry (UPLC-PDA-MS/MS). The results showed that all pigments presented a very high antioxidant activity and good stability under exposure to UV light. However, except for a fraction of the A. agilis 50cyt pigment, they were shown to be phototoxic. A total of 18 different carotenoids were identified from 23 that were separated on a C18 column. The C50 carotenes bacterioruberin and decaprenoxanthin (including its variations) were confirmed for A. agilis 50cyt and A. psychrochitiniphilus 366, respectively. All-trans-bacterioruberin was identified as the pigment that did not express phototoxic activity in the 3T3 NRU PT assay (MPE < 0.1). Zeaxanthin, β-cryptoxanthin, β-carotene, and phytoene were detected in Z. laminarie 465. In conclusion, carotenoids identified in this work from Antarctic bacteria open perspectives for their further biotechnological application towards a more sustainable and environmentally friendly way of pigment exploitation.
KeywordsAntarctic pigments, Psychrophilic bacteria Arthrobacter Zobellia Antioxidant Carotenoids
The authors are grateful to Fundação de Amparo à Pesquisa do Estado de São Paulo–FAPESP for financial funding (process numbers 2014/17936-1, 2016/05640-6, and 2017/21790-0). The MycoAntar Project (CNPq) and the Brazilian Antarctic Program are also acknowledged for making the sampling feasible in the OPERANTAR XXXIII (summer 2014/2015) and OPERANTAR XXXIV (summer 2015/2016). We also would like to thank Dr. Marcos Eberlin and Dr. Fabio Neves from ThoMSon Mass Spectrometry Laboratory in UNICAMP for the analytical chemistry training.
This study was funded by the São Paulo Research Foundation–FAPESP (grant numbers 2014/17936-1, 2016/05640-6, and 2017/21790-0).
Compliance with Ethical Standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of Interest
The authors declare that they have no conflict of interest.
- Bowman JP, McCammon SA, Brown MV et al (1997) Diversity and association of psychrophilic bacteri in Antarctic sea ice. Appl Environ Microbiol 63:3068–3078Google Scholar
- Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free radical method to evaluate antioxidant activity. LWT -Food Sci Technol 28:25–30Google Scholar
- Britton G, Pfander H, Liaaen-Jensen S (2004) Carotenoids handbook. Birkhäuser Verlag, Basel BostonGoogle Scholar
- Ceridono M, Tellner P, Bauer D, Barroso J, Alépée N, Corvi R, de Smedt A, Fellows MD, Gibbs NK, Heisler E, Jacobs A, Jirova D, Jones D, Kandárová H, Kasper P, Akunda JK, Krul C, Learn D, Liebsch M, Lynch AM, Muster W, Nakamura K, Nash JF, Pfannenbecker U, Phillips G, Robles C, Rogiers V, van de Water F, Liminga UW, Vohr HW, Wattrelos O, Woods J, Zuang V, Kreysa J, Wilcox P (2012) The 3T3 neutral red uptake phototoxicity test: practical experience and implications for phototoxicity testing - the report of an ECVAM-EFPIA workshop. Regul Toxicol Pharmacol 63:480–488CrossRefGoogle Scholar
- ECVAM DB-ALM (2008) 3T3 Neutral Red Uptake (NRU) phototoxicity assay. DB-ALM Protocol n ° 78, pp 1–19Google Scholar
- Ehling-Schulz M, Scherer S (1999) UV protection in cyanobacteria. Eur J Phycol 34(4):329–323Google Scholar
- Giuffrida D, Sutthiwong N, Dugo P et al (2016) Characterisation of the C50 carotenoids produced by strains of the cheese-ripening bacterium Arthrobacter arilaitensis. Int Dairy J 55:10–16Google Scholar
- Harm W (1980) Biological effects of ultraviolet radiation. Cambridge University Press, LondonGoogle Scholar
- Holzhütter HG (1997) A general measure of in vitro phototoxicity derived from pairs of dose-response curves and its use for predicting the in vivo phototoxicity of chemicals. ATLA Altern Lab Anim 25:445–462Google Scholar
- ICH (1996) Photostability Testing of New Drug Substances and Products Q1B, International Conference on Harmonisation, IFPMA, GenevaGoogle Scholar
- Kleinig H, Heumann W, Meister W, Englert G (1977) Carotenoids of rhizobia. I. New carotenoids from rhizobium lupini. Helv Chim Acta 60:254–258. Google Scholar
- Kuhlman KR, Allenbach LB, Ball CL, Fusco WG, la Duc MT, Kuhlman GM, Anderson RC, Stuecker T, Erickson IK, Benardini J, Crawford RL (2005) Enumeration, isolation, and characterization of ultraviolet (UV-C) resistant bacteria from rock varnish in the Whipple Mountains, California. Icarus 174:585–595CrossRefGoogle Scholar
- Link L, Sawyer J, Venkateswaran K, Nicholson W (2004) Extreme spore UV resistance of Bacillus pumilus isolates obtained from an ultraclean spacecraft assembly facility. Microb Ecol:159–163Google Scholar
- Margesin R, Schinner F, Marx JC, Gerday C (2008) Psychrophiles, from Biodiversity to Biotechnology. Springer, BerlinGoogle Scholar
- Mathews-Roth MM (1987) Photoprotection by carotenoids. Fed Proc 46:1890–1893Google Scholar
- OECD (2004) OECD guidelines for the testing of chemicals. Test:1–21. https://doi.org/10.1787/9789264203785-en
- Prior RL, Hoang H, Gu L, Wu X, Bacchiocca M, Howard L, Hampsch-Woodill M, Huang D, Ou B, Jacob R (2003) Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORACFL)) of plasma and other biological and food samples. J Agric Food Chem 51:3273–3279CrossRefGoogle Scholar
- Sajilata MG, Singhal RS, Kamat MY (2008) The carotenoid pigment zeaxanthin - a review. Compr Rev Food Sci Food Saf:29–49Google Scholar
- Schwender J, Seemann M, Lichtenthaler HK, Rohmer M (1996) Biosynthesis of isoprenoids (carotenoids, sterols, prenyl side-chains of chlorophylls and plastoquinone) via a novel pyruvate/glyceraldehyde 3-phosphate non-mevalonate pathway in the green alga Scenedesmus obliquus. Biochem J 316(Pt 1):73–80CrossRefGoogle Scholar
- Shaath N (2007) SPF boosters & photostability of ultraviolet filters. Happi 10:77–83Google Scholar
- Silva TR, Duarte AWF, Passarini MRZ, Ruiz ALTG, Franco CH, Moraes CB, de Melo IS, Rodrigues RA, Fantinatti-Garboggini F, Oliveira VM (2018) Bacteria from Antarctic environments: diversity and detection of antimicrobial, antiproliferative, and antiparasitic activities. Polar Biol 41:1505–1519CrossRefGoogle Scholar
- Spielmann H, Balls M, Dupuis J, Pape WJ, Pechovitch G, de Silva O, Holzhütter HG, Clothier R, Desolle P, Gerberick F, Liebsch M, Lovell WW, Maurer T, Pfannenbecker U, Potthast JM, Csato M, Sladowski D, Steiling W, Brantom P (1998) The international EU/COLIPA in vitro phototoxicity validation study: results of phase II (blind trial). Part 1: the 3T3 NRU phototoxicity test. Toxicol In Vitro 12:305–327CrossRefGoogle Scholar
- Wynn-Williams DD, Edwards HGM (2002) Environmental UV radiation: biological strategies for protection and avoidance. Astrobiol Quest Cond Life:245–260Google Scholar
- Wynn-Williams DD, Newton EM, Edwards HGM (2001) The role of habitat structure for biomolecule integrity and microbial survival under extreme environmental stress in Antarctica (and Mars?): ecology and technology. In Exo-/astro-biology (vol 496. pp. 225-237)Google Scholar