A Report on Finding a New Peptide Aldehyde from Cyanobacterium Nostoc sp. Bahar M by LC-MS and Marfey’s Analysis

Document Type : Research Paper


1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Division of Microbiology, Department of Food and Environmental Sciences, University of Helsinki, Finland


Background: Cyanobacteria have a worldwide distribution in the terrestrial habitats, occurring predominantly on the surface of the soils, stones, rocks, and trees, practically in moist, neutral or alkaline aeries. The unique natural and bioactive compounds from cyanobacteria with various biological activities and an extensive range of chemical classes have a significant capability for expansion of the pharmaceuticals and other biomedical purposes.
Objectives: Regardless of the progresses in our knowledge on cyanobacteria, however, cyanobacteria are still viewed as an unexplored source of potential drugs. In this study presence of bioactive compounds among the cyanobacteria culture collection of Iran, where a wide variety of strains can be found, was investigated.
Material and Methods: We explored one Nostoc strain isolated from rice fields in Golestan province of northern Iran for searching for novel products. The chemical construction of the new bioactive compound was clarified by application of liquid chromatography-mass spectrometer (LC-MS) and Marfey’s analysis of the degradation products.
Results: We found a novel peptide aldehyde compound from a hydrophilic extract of the Nostoc sp. Bahar_M, which is composed of the three subunits, 2-hydroxy-4-(4-hydroxyphenyl) butanoic acid (Hhpba), L-Ile, and L-argininal. According to the structural information, we predicted that the novel peptide-aldehyde compound probably to be trypsin inhibitors.
Conclusions: Results demonstrated that terrestrial cyanobacteria are a promissing resource of bioactive natural products.


Main Subjects

1.           Dembitsky VM, Rezanka T. Metabolites produced by nitrogen-fixing Nostoc species. Folia Microbiol (Praha). 2005;50(5):363-391. doi: 10.1007/BF02931419 pmid: 16475497
2.           Ehrenreich IM, Waterbury JB, Webb EA. Distribution and diversity of natural product genes in marine and freshwater cyanobacterial cultures and genomes. Appl Environ Microbiol. 2005;71(11):7401-7413. doi: 10.1128/AEM.71.11.7401-7413.2005 pmid: 16269782
3.           Dittmann E, Neilan BA, Borner T. Molecular biology of peptide and polyketide biosynthesis in cyanobacteria. Appl Microbiol Biotechnol. 2001;57(4):467-473. doi: 10.1007/s002530100810 pmid: 11764765
4.           Bui HT, Jansen R, Pham HT, Mundt S. Carbamidocyclophanes A-E, chlorinated paracyclophanes with cytotoxic and antibiotic activity from the Vietnamese cyanobacterium Nostoc sp. J Nat Prod. 2007;70(4):499-503. doi: 10.1021/np060324m pmid: 17311455
5.           Kajiyama S-i, Kanzaki H, Kawazu K, Kobayashi A. Nostofungicidine, an antifungal lipopeptide from the field-grown terrestrial blue-green alga Nostoc commune. Tetrahedron Lett. 1998;39(22):3737-3740. doi: 10.1016/s0040-4039(98)00573-5
6.           Jaki B, Heilmann J, Sticher O. New antibacterial metabolites from the cyanobacterium Nostoc commune(EAWAG 122b). J Nat Prod. 2000;63(9):1283-1285. doi: 10.1021/np000033s pmid: 11000038
7.           Kanekiyo K, Lee JB, Hayashi K, Takenaka H, Hayakawa Y, Endo S, et al. Isolation of an antiviral polysaccharide, nostoflan, from a terrestrial cyanobacterium, Nostoc flagelliforme. J Nat Prod. 2005;68(7):1037-1041. doi: 10.1021/np050056c pmid: 16038544
8.           Murakami M, Ishida K, Okino T, Okita Y, Matsuda H, Yamaguchi K. Aeruginosins 98-A and B, trypsin inhibitors from the blue-green alga Microcystis aeruginosa (NIES-98). Tetrahedron Lett. 1995;36(16):2785-2788. doi: 10.1016/0040-4039(95)00396-t
9.           Fewer DP, Jokela J, Rouhiainen L, Wahlsten M, Koskenniemi K, Stal LJ, et al. The non-ribosomal assembly and frequent occurrence of the protease inhibitors spumigins in the bloom-forming cyanobacterium Nodularia spumigena. Mol Microbiol. 2009;73(5):924-937. doi: 10.1111/j.1365-2958.2009.06816.x pmid: 19691450
10.        Pluotno A, Carmeli S. Banyasin A and banyasides A and B, three novel modified peptides from a water bloom of the cyanobacterium Nostoc sp. Tetrahedron. 2005;61(3):575-583. doi: 10.1016/j.tet.2004.11.016
11.        von Elert E, Oberer L, Merkel P, Huhn T, Blom JF. Cyanopeptolin 954, a chlorine-containing chymotrypsin inhibitor of Microcystis aeruginosa NIVA Cya 43. J Nat Prod. 2005;68(9):1324-1327. doi: 10.1021/np050079r pmid: 16180807
12.        Zafrir-Ilan E, Carmeli S. Eight novel serine proteases inhibitors from a water bloom of the cyanobacterium Microcystis sp. Tetrahedron. 2010;66(47):9194-9202. doi: 10.1016/j.tet.2010.09.067
13.        Taori K, Paul VJ, Luesch H. Kempopeptins A and B, serine protease inhibitors with different selectivity profiles from a marine cyanobacterium, Lyngbya sp. J Nat Prod. 2008;71(9):1625-1629. doi: 10.1021/np8002172 pmid: 18693761
14.        Reshef V, Carmeli S. Protease inhibitors from a water bloom of the cyanobacterium Microcystis aeruginosa. Tetrahedron. 2001;57(14):2885-2894. doi: 10.1016/s0040-4020(01)00141-7
15.        Becher PG, Baumann HI, Gademann K, Jüttner F. The cyanobacterial alkaloid nostocarboline: an inhibitor of acetylcholinesterase and trypsin. J Appl Phycol. 2008;21(1):103-110. doi: 10.1007/s10811-008-9335-3
16.        Reshef V, Carmeli S. New microviridins from a water bloom of the cyanobacterium Microcystis aeruginosa. Tetrahedron. 2006;62(31):7361-7369. doi: 10.1016/j.tet.2006.05.028
17.        Rippka R, Stanier RY, Deruelles J, Herdman M, Waterbury JB. Generic Assignments, Strain Histories and Properties of Pure Cultures of Cyanobacteria. Microbiology. 1979;111(1):1-61. doi: 10.1099/00221287-111-1-1
18.        Kaushik BD. Laboratory methods for blue-green algae. New Delhi: Associated Publishing co.; 1987.
19.        Desikachary TV. Cyanophyta. New Delhi: Indian council of Agricultural Research; 1959.
20.        Nowruzi B, Khavari-Nejad R-A, Nejadsattari T, Sivonen K, Fewer D. A proposal for the unification of two cyanobacterial strains of Nostoc as the same species. Rostaniha. 2017;17(2):161-172.
21.        Leikoski N, Fewer DP, Jokela J, Wahlsten M, Rouhiainen L, Sivonen K. Highly diverse cyanobactins in strains of the genus Anabaena. Appl Environ Microbiol. 2010;76(3):701-709. doi: 10.1128/AEM.01061-09 pmid: 20008171
22.        Staub R. Ernährungsphysiologisch-autökologische Untersuchungen an der planktischen BlaualgeOscillatoria rubescens DC. Schweizerische Zeitschrift für Hydrologie. 1961;23(1):82-198. doi: 10.1007/bf02505618
23.        Kotai J. Instructions for preparation of modified nutrient solution Z8 for algae. Norwegian Inst Water Res. 1972;11(69):5.
24.        Niva A. Estimation of algal growth potential. Norwegian Inst Water Res. 1976;11(82):2-25.
25.        Hoffmann D, Hevel JM, Moore RE, Moore BS. Sequence analysis and biochemical characterization of the nostopeptolide A biosynthetic gene cluster from Nostoc sp. GSV224. Gene. 2003;311:171-180. doi: 10.1016/S0378-1119(03)00587-0 pmid: 12853152
26.        Jokela J, Herfindal L, Wahlsten M, Permi P, Selheim F, Vasconcelos V, et al. A novel cyanobacterial nostocyclopeptide is a potent antitoxin against microcystins. Chembiochem. 2010;11(11):1594-1599. doi: 10.1002/cbic.201000179 pmid: 20575133
27.        Nowruzi B, Haghighat S, Fahimi H, Mohammadi E. Nostoc cyanobacteria species: a new and rich source of novel bioactive compounds with pharmaceutical potential. J Pharm Health Serv Res. 2018;9(1):5-12. doi: 10.1111/jphs.12202
28.        B. N, Khavari-Nejad RA, Sivonen K, Kazemi B, Najafi F, Nejadsattari T. A gene expression study on strains of Nostoc (Cyanobacteria) revealing antimicrobial activity under mixotrophic conditions. African J Biotech. 2012;11(51):11296-11308. doi: 10.5897/ajb11.4129
29.        Chlipala GE, Mo S, Orjala J. Chemodiversity in freshwater and terrestrial cyanobacteria - a source for drug discovery. Curr Drug Targets. 2011;12(11):1654-1673. doi: 10.2174/138945011798109455 pmid: 21561419
30.        Burja AM, Banaigs B, Abou-Mansour E, Grant Burgess J, Wright PC. Marine cyanobacteria—a prolific source of natural products. Tetrahedron. 2001;57(46):9347-9377. doi: 10.1016/s0040-4020(01)00931-0
31.        Tan LT. Bioactive natural products from marine cyanobacteria for drug discovery. Phytochemistry. 2007;68(7):954-979. doi: 10.1016/j.phytochem.2007.01.012 pmid: 17336349
32.        Matthew S, Ratnayake R, Becerro MA, Ritson-Williams R, Paul VJ, Luesch H. Intramolecular modulation of serine protease inhibitor activity in a marine cyanobacterium with antifeedant properties. Mar Drugs. 2010;8(6):1803-1816. doi: 10.3390/md8061803 pmid: 20631871
33.        Tooming-Klunderud A, Rohrlack T, Shalchian-Tabrizi K, Kristensen T, Jakobsen KS. Structural analysis of a non-ribosomal halogenated cyclic peptide and its putative operon from Microcystis: implications for evolution of cyanopeptolins. Microbiology. 2007;153(Pt 5):1382-1393. doi: 10.1099/mic.0.2006/001123-0 pmid: 17464052
34.        Kalaitzis JA, Lauro FM, Neilan BA. Mining cyanobacterial genomes for genes encoding complex biosynthetic pathways. Nat Prod Rep. 2009;26(11):1447-1465. doi: 10.1039/b817074f pmid: 19844640
35.        Jaspars M, Lawton LA. Cyanobacteria - a novel source of pharmaceuticals. Curr Opin Drug Discov Devel. 1998;1(1):77-84. pmid: 19649793
36.        Ahmadi Moghadam A, Nowruzi B. A new report of n fixation by two species of cyanobacteria. Iran J Sci Technol. 2008;32(2):147-151.
37.        Singh S, Kate BN, Banerjee UC. Bioactive compounds from cyanobacteria and microalgae: an overview. Crit Rev Biotechnol. 2005;25(3):73-95. doi: 10.1080/07388550500248498 pmid: 16294828
38.        Sivonen K, Börner T. Bioactive compounds produced by cyanobacteria; The cyanobacteria: molecular biology, genomics and evolution. Norfolk: Caister academic press; 2008. 159-197 p.
39.        Gademann K, Portmann C. Secondary Metabolites from Cyanobacteria: Complex Structures and Powerful Bioactivities. Curr Org Chem. 2008;12(4):326-341. doi: 10.2174/138527208783743750
40.        Golakoti T, Yoshida WY, Chaganty S, Moore RE. Isolation and Structures of Nostopeptolides A1, A2 and A3 from the Cyanobacterium Nostoc sp. GSV224. Tetrahedron. 2000;56(46):9093-9102. doi: 10.1016/s0040-4020(00)00764-x
41.        Golakoti T, Yoshida WY, Chaganty S, Moore RE. Isolation and structure determination of nostocyclopeptides A1 and A2 from the terrestrial cyanobacterium Nostoc sp. ATCC53789. J Nat Prod. 2001;64(1):54-59. doi: 10.1021/np000316k pmid: 11170666
42.        Luesch H, Hoffmann D, Hevel JM, Becker JE, Golakoti T, Moore RE. Biosynthesis of 4-methylproline in cyanobacteria: cloning of nosE and nosF genes and biochemical characterization of the encoded dehydrogenase and reductase activities. J Org Chem. 2003;68(1):83-91. doi: 10.1021/jo026479q pmid: 12515465
43.        Becker JE, Moore RE, Moore BS. Cloning, sequencing, and biochemical characterization of the nostocyclopeptide biosynthetic gene cluster: molecular basis for imine macrocyclization. Gene. 2004;325:35-42. doi: 10.1016/j.gene.2003.09.034 pmid: 14697508
44.        Hunsucker SW, Klage K, Slaughter SM, Potts M, Helm RF. A preliminary investigation of the Nostoc punctiforme proteome. Biochem Biophys Res Commun. 2004;317(4):1121-1127. doi: 10.1016/j.bbrc.2004.03.173 pmid: 15094385
45.        Fewer DP, Jokela J, Paukku E, Osterholm J, Wahlsten M, Permi P, et al. New structural variants of aeruginosin produced by the toxic bloom forming cyanobacterium Nodularia spumigena. PLoS One. 2013;8(9):e73618. doi: 10.1371/journal.pone.0073618 pmid: 24040002
46.        Liu L, Jokela J, Wahlsten M, Nowruzi B, Permi P, Zhang YZ, et al. Nostosins, Trypsin Inhibitors Isolated from the Terrestrial Cyanobacterium Nostoc sp. Strain FSN. J Nat Prod. 2014;77(8):1784-1790. doi: 10.1021/np500106w pmid: 25069058
47.        Liu L, Jokela J, Herfindal L, Wahlsten M, Sinkkonen J, Permi P, et al. 4-Methylproline guided natural product discovery: co-occurrence of 4-hydroxy- and 4-methylprolines in nostoweipeptins and nostopeptolides. ACS Chem Biol. 2014;9(11):2646-2655. doi: 10.1021/cb500436p pmid: 25203327
48.        Shim SH, Chlipala G, Orjala J. Isolation and structure determination of a proteasome inhibitory metabolite from a culture of Scytonema hofmanni. J Microbiol Biotechnol. 2008;18(10):1655-1658. pmid: 18955814
49.        Turk B. Targeting proteases: successes, failures and future prospects. Nat Rev Drug Discov. 2006;5(9):785-799. doi: 10.1038/nrd2092 pmid: 16955069
50.        Smith JL, Boyer GL, Zimba PV. A review of cyanobacterial odorous and bioactive metabolites: Impacts and management alternatives in aquaculture. Aquaculture. 2008;280(1-4):5-20. doi: 10.1016/j.aquaculture.2008.05.007
51.        Welker M, von Dohren H. Cyanobacterial peptides - nature's own combinatorial biosynthesis. FEMS Microbiol Rev. 2006;30(4):530-563. doi: 10.1111/j.1574-6976.2006.00022.x pmid: 16774586
52.        Baumann HI, Keller S, Wolter FE, Nicholson GJ, Jung G, Sussmuth RD, et al. Planktocyclin, a cyclooctapeptide protease inhibitor produced by the freshwater cyanobacterium Planktothrix rubescens. J Nat Prod. 2007;70(10):1611-1615. doi: 10.1021/np0700873 pmid: 17935298
53.        Richman DD. HIV therapeutics. Science. 1996;272(5270):1886-1888. doi: 10.1126/science.272.5270.1886 pmid: 8658159
54.        Mohammadian M, Farzampanah L, Behtash-oskouie A, Majdi S, Mohseni G, Imandar M, et al. A biosensor for detect nitrite (NO2-) and hydroxylamine (nh2oh) by using of hydroxylamine oxidase and modified electrode with ZnO nanoparticles. Int J Electrochem Sci. 2013;8(9):11215-11227.
55.        Borowitzka MA. Pharmaceuticals and agrochemicals from microalgae. In: Cohen Z, editor. Chemicals from microalgae: Taylor and Francis Ltd; 1999. p. 313-352.
56.        Koivunen E, Itkonen O, Halila H, Stenman UH. Cyst fluid of ovarian cancer patients contains high concentrations of trypsinogen-2. Cancer Res. 1990;50(8):2375-2378. pmid: 2180568
57.        Nyberg P, Ylipalosaari M, Sorsa T, Salo T. Trypsins and their role in carcinoma growth. Exp Cell Res. 2006;312(8):1219-1228. doi: 10.1016/j.yexcr.2005.12.024 pmid: 16457812
58.        Wang Y, Luo W, Reiser G. Trypsin and trypsin-like proteases in the brain: proteolysis and cellular functions. Cell Mol Life Sci. 2008;65(2):237-252. doi: 10.1007/s00018-007-7288-3 pmid: 17965832