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Mycosporine-Like Amino Acids from Biological Integuments of Historical Monuments

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Mycosporine-Like Amino Acids from Biological Integuments of Historical Monuments Int.J.Curr.Microbiol.App.Sci (2016) 5(8): 30-41 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 5 Number 8 (2016) pp. 30-41 Journal homepage: http://www.ijcmas.com Original Research Article http://dx.doi.org/10.20546/ijcmas.2016.508.004 Mycosporine-like Amino Acids from Biological Integuments of Historical Monuments Arun S. Sonker, Richa, Jainendra Pathak, Rajneesh, Vinod K. Kannaujiya and Rajeshwar P. Sinha* Laboratory of Photobiology and Molecular Microbiology, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi-221005, India *Corresponding author ABSTRACT Biological crusts collected from seven historical monuments in and around K e yw or ds Varanasi, India, were screened for the presence of photoprotective mycosporine- like amino acids (MAAs). Nine strains of cyanobacterial genera such as Lyngbya Biological films, Carotenoids, sp., Nostoc sp., Anabaena sp., Scytonema sp., Phormidium sp., Westiellopsis sp., Chlorophyll a, Aphanocapsa sp., Hapalosiphon sp. and Aphanothece sp. were found to be present in the samples collected from various monuments apart from other group of Cyanobacteria, Historical organisms. Cyanobacteria, the dominant population growing on all the monuments monuments, had a maximum diversity of 5 species in Brahaspati temple, 4 species in Ramnagar Mycosporine -like fort and LalKhan`s tomb, 3 species in Manikarnika ghat and Sanskrit University amino acids. and only 1 species in Sarnath and Bharat Mata temple. Pigment profile of the crusts Article Info from the seven monuments showed peaks at 665, 470, 310 and 386 nm that correspond to the presence of chlorophyll a, carotenoids, MAAs and scytonemin Accepted: respectively. High content of cholorophyll a was recorded in the crusts from 06 July 2016 Sarnath and the Brahaspati temple, whereas carotenoids content was higher in the Available Online: crust sample of LalKhan`s tomb. In all the collected samples from various 10 August 2016 monuments, photoprotective MAAs were found to be predominant than the photosynthetic pigments (chlorophyll a and carotenoids). Introduction early Earth, when there was no ozone layer Cyanobacteria are Gram-negative (Schopf, 2000; Hedges et al., 2001; Häder et prokaryotes having a cosmopolitan al., 2015). The ancient monument all over distribution ranging from hot-springs to the the world are deteriorating due to the Arctic and Antarctic regions (Stanier and deposition of cyanobacteria, resulting in the Cohen-Bazire, 1977). They were the first loss of basic structure and creates pits, photosynthetic oxygen-evolving prokaryotes cracks and fissures (Ortega-Calvo et al., which appeared during the Precambrian era 1991; Dhami et al., 2014). (Brocks et al., 1999). Fossil records, organic biomarkers and genomic sequence analyses On ancient monuments, cyanobacteria indicate the presence of cyanobacteria on the played an important role as pioneer for 30 Int.J.Curr.Microbiol.App.Sci (2016) 5(8): 30-41 establishing life on bare inorganic rocks and artistic work and are important part of produced considerable biomass. Most of glorious traditions and rich cultural heritage these cyanobacterial crusts growing on the of Varanasi. ancient monuments were colonial or filamentous and occurred in association with Photoprotective MAAs are <400 Da, other algae, fungi, bacteria, lichen and moss colorless, water-soluble compounds Protonema (Lüttge et al., 1995). Several composed of a cyclohexenone or studies on the cyanobacterial crust residing cyclohexenimine chromophore conjugated on the ancient monuments have been done with the nitrogen substituent of an amino in South-West USA and Mexico acid or its imino alcohol (Carreto et al., (Friedmann, 1972), monuments of Northern 2005; Richa and Sinha, 2015) and have Transvall, South Africa, marbles of absorption maxima in the range of 310-362 Parthenon (Acropolis-Athens) of Greece nm. Generally, the ring system contains a (Anagnostidis et al., 1983), Roman frescoes glycine subunit linked to the third carbon (Grilli-Caiola et al., 1987; Albertano and atom. Some MAAs also contains sulfate Grilli-Caiola, 1989), Inferniglio cave of Italy esters or glycosidic linkages through the (Abdelahad, 1989), Lund cathedral of imine substituents. Differences between the Sweden (Ortega-Calvoet al., 1991), Roman absorption spectra of MAAs are due to the Necropolis (Albertano et al., 1994) and attached side groups and nitrogen Goldengate highlands National Park, South substituent. The biosynthesis of MAAs has Africa (Wessels and Büdel,1989) etc. been predicted to occur via the first part of the shikimate pathway but concluding In Varanasi (India) region, so far no prior evidences are lacking. It has been found that studies have been done on the presence of 3-dehydroquinate, which is formed in the photoprotective MAAs in the crust forming centre of the shikimate pathway, acts as a cyanobacteria of the monuments. Some of precursor for the synthesis of fungal these monuments are situated in the warm mycosporines and MAAs via gadusols temperate region having a very suitable (Shick et al., 2002). condition for the growth and development of cyanobacteria which is responsible for the MAAs provide protection from UV defragmentation of the monuments. Though radiation not only in their producers but also such problems are important in humid and to primary and secondary consumers tropical climates, there is little information through the food chain. MAAs has been about the cyanobacteria growing on the reported in diverse organisms, such as, monuments of India (Tripathy et al., 1999; bacteria, cyanobacteria, macroalgae, Pattanaik and Adhikary, 2002). Some phytoplankton and various animals such as historical monuments of Varanasi such as, arthropods, rotifers, molluscs, fishes, Sarnath temple is about three hundred years cnidarians, tunicates, poriferans, nematodes, old and a holy place for Buddhists, LalKha echinodermates, platythelminthes, ka Rauza build by Mughal emperor is polychaetes, bryozoans and protozoans situated at the bank of sacred river Ganges, (Sinha et al., 2007; Kannaujiya et al., 2014; Sanskrit University is about 150 years old, Pathak et al., 2015), but not in animals as Ramnagar fort was manufactured by the they lack the shikimate pathway, but these King of Varanasi about five hundred years compounds may be accumulated either via ago and Manikarnika ghat temple is about the food chain or synthesized by their 600 hundred years old, have wonderful symbiotic algal partner (Shick and Dunlap, 31 Int.J.Curr.Microbiol.App.Sci (2016) 5(8): 30-41 2002). Presently, about 22 MAAs have been 48 h. To examine the presence of reported from terrestrial, freshwater and cyanobacteria, a pinch of the rehydrated marine organisms. MAAs have high molar crusts were observed under compound extinction coefficient which favours them as microscope (Getneroptik KFS4 (I) 5582) strong photoprotectant. In addition, to their and photographed using Dewinter image photoprotective role in various organisms, microscope fitted with digital camera. MAAs are also of immense importance for Various cyanobacteria, such as, Anabaena humans as these compounds have been sp., Nostoc sp., Lyngbya sp. and Scytonema found to effectively block thymine dimer sp. etc. were found to be present in the formation by UVR in vitro, have antioxidant collected crusts. as well as growth stimulation activity in human cells and were recently found to Mycosporine-like amino acids (MAAs) reduce UV-induced aging (Oyamada et al., extraction and spectroscopic analysis 2008; Singh et al., 2010). In the present study, for the first time, we have reported For extraction of MAAs, equal amount of the diversity of cyanobacteria and the crust (0.5 mg) collected from various presence of photoprotective MAAs from the monuments were dissolved in 2 mL of exposed surfaces of different monuments of 100% methanol (HPLC-grade) and Varanasi, India. incubated at 4 °C for overnight. The methanol extracts were centrifuged at Materials and Methods 10,000 rpm for 10 min, and the supernatant was subjected to spectroscopic analysis Collection sites between 250-700 nm wavelengths, using a double beam spectrophotometer (UV-VIS The cyanobacterial samples were collected 2900, Hitachi, Japan). The raw spectra from various monuments such as, LalKhan`s (peaks) were analyzed using UV Probe tomb, Manikarnika ghat, Brahaspati temple, version software (Hitachi, Japan). Sarnath, Ramnagar fort, Sanskrit University and Bharat Mata temple. These monuments Purification and estimation of MAAs are situated between 25° 20' 0" North, 83° 0' 0"East in Varanasi district of Uttar Pradesh, After initial characterization of MAAs by India. These historical places are very spectroscopic analysis, methanolic extracts popular and some are protected by the were evaporated to dryness at 450C, and the Archaeological survey of India. Since the dried product was dissolved in 1ml Milli Q cyanobacterial colonization occurs in the water in a microcentrifuge tube. After upper surface of the crust, we have collected adding a few drops of chloroform, the the crust growing over these monuments by suspension was subjected to centrifugation, a non-destructive double layered adhesive and the water phase was carefully tape method and then transferred to pre- transferred into a fresh microcentrifuge tube sterilized screw-cap bottles and transported to remove contaminating lipophillic to the laboratory for further analyses. compounds. Identification of organisms Finally, the resulting suspension
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