Abstract
Herein we report the isolation of a novel actinomycete, strain MCCB 268T, from the sediment sample collected from a high Arctic fjord Kongsfjorden. MCCB 268T showed greater than 97% 16S rRNA gene sequence similarity with those of Pseudonocardia konjuensis LM 157T (98.06%), Pseudonocardia soli NW8-21 (97.22%) Pseudonocardia endophytica YIM 56035 (97.08%) and Pseudonocardia nantongensis KLBMP 1282 (97.34%) showing that the strain should be assigned to the genus Pseudonocardia. DNA–DNA hybridization with Pseudonocardia konjuensis LM 157T showed only 41.5% relatedness to strain MCCB 268T. The whole genome of the strain MCCB 268T was sequenced. Whole-genome average nucleotide identity, dDDH (%) and genome tree analysis demonstrated that strain significantly differed from other Pseudonocardia species. The G + C content was 70.5 mol%. MCCB 268T exhibited in vitro cytotoxicity and through bioassay guided fractionation followed by HPLC separation a cytotoxic compound (I) was isolated. The compound (I) was identified as 1-acetyl-β-carboline through NMR spectra and high-resolution mass spectrometry. Compound (I) showed cytotoxicity against lung cancer cell line and mode of anticancer activity was found to be through the induction of apoptosis. Based on the genotypic and phenotypic features, MCCB 268T ought to be classified as a novel species under the genus Pseudonocardia for which the name Pseudonocardia cytotoxica sp. nov. is proposed (= CCUG72333T = JCM32718T).
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References
Abramovitch RA, Spenser ID (1964) The carbolines. Adv Heterocycl Chem 3:79–207
Allen JRF, Holmstedt BR (1980) The simple β-carboline alkaloids. Phytochemistry 19:1573–1582. https://doi.org/10.1016/S0031-9422(00)83773-5
Brownlee KA, Finney DJ, Tattersfield F (1952) Probit analysis: a statistical treatment of the sigmoid response curve. J Am Stat Assoc 47:687. https://doi.org/10.2307/2280787
Cao R, Peng W, Wang Z, Xu A (2007) beta-Carboline alkaloids: biochemical and pharmacological functions. Curr Med Chem 14:479–500
Chan YY, Juang SH, Huang GJ et al (2014) The constituents of Michelia compressa var. Formosana and their bioactivities. Int J Mol Sci 15:10926–10935. https://doi.org/10.3390/ijms150610926
Chen HH, Qin S, Li J et al (2009) Pseudonocardia endophytica sp. nov., isolated from the pharmaceutical plant Lobelia clavata. Int J Syst Evol Microbiol 59:559–563. https://doi.org/10.1099/ijs.0.64740-0
Cheng HR, Jiang N (2006) Extremely rapid extraction of DNA from bacteria and yeasts. Biotechnol Lett 28:55–59. https://doi.org/10.1007/s10529-005-4688-z
Chun J, Kim BK, Lim Y-W et al (2007) EzTaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences. Int J Syst Evol Microbiol 57:2259–2261. https://doi.org/10.1099/ijs.0.64915-0
Collins MD, Pirouz T, Goodfellow M, Minnikin DE (1977) Distribution of menaquinones in actinomycetes and corynebacteria. J Gen Microbiol 100:221–230. https://doi.org/10.1099/00221287-100-2-221
De Ley J, Cattoir H, Reynaerts A (1970) The quantitative measurement of DNA hybridization from renaturation rates. Eur J Biochem 12:133–142. https://doi.org/10.1111/j.1432-1033.1970.tb00830.x
de Meester C (1995) Genotoxic potential of beta-carbolines: a review. Mutat Res 339:139–153
Dhaneesha M, Benjamin Naman C, Krishnan KP et al (2017) Streptomyces artemisiae MCCB 248 isolated from Arctic fjord sediments has unique PKS and NRPS biosynthetic genes and produces potential new anticancer natural products. Biotechnology 7:32. https://doi.org/10.1007/s13205-017-0610-3
Dhaneesha M, Hasin O, Sivakumar KC et al (2019) DNA binding and molecular dynamic studies of polycyclic tetramate macrolactams (PTM) with potential anticancer activity isolated from a sponge-associated Streptomyces zhaozhouensis subsp. mycale subsp. nov. Mar Biotechnol 21:124–137. https://doi.org/10.1007/s10126-018-9866-9
Felsenstein J (1985) Phylogenies and the comparative method. Am Nat 125:1–15. https://doi.org/10.1086/284325
Fitch WM (1971) Toward defining the course of evolution: minimum change for a specific tree topology. Syst Biol 20:406–416. https://doi.org/10.1093/sysbio/20.4.406
Fu P, Johnson M, Chen H et al (2014) Carpatamides A-C, cytotoxic arylamine derivatives from a marine-derived streptomyces sp. J Nat Prod 77:1245–1248. https://doi.org/10.1021/np500207p
Gonzalez JM (2002) Brief report A fluorimetric method for the estimation of G + C mol % content in microorganisms by thermal denaturation temperature. Environ Microbiol 4:770–773
Gosink JJ, Woese CR, Staley JT (1998) Polaribacter gen. nov., with three new species, P. irgensii sp. nov., P. franzmannii sp. nov. and P. filamentus sp. nov., gas vacuolate polar marine bacteria of the Cytophaga-Flavobacterium-Bacteroides group and reclassification of ’Flectobacillus glomera. Int J Syst Bacteriol 48:223–235. https://doi.org/10.1099/00207713-48-1-223
Ishida J, Wang HK, Bastow KF et al (1999) Antitumor agents 201. Cytotoxicity of harmine and beta-carboline analogs. Bioorg Med Chem Lett 9:3319–3324
Kelly KL (1964) Inter-society color council—national bureau of standards color name charts illustrated with centroid colors. US Government Printing Office, Washington
Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120. https://doi.org/10.1007/BF01731581
Klappenbach JA, Goris J, Vandamme P et al (2007) DNA–DNA hybridization values and their relationship to whole-genome sequence similarities. Int J Syst Evol Microbiol 57:81–91. https://doi.org/10.1099/ijs.0.64483-0
Knoblauch C, Sahm K, Jorgensen BB (1999) Psychrophilic sulfate-reducing bacteria isolated from permanently cold Arctic marine sediments: description of Desulfofrigus oceanense gen. nov., sp. nov., Desulfofrigus fragile sp. nov., Desulfofaba gelida gen. nov., sp. nov. Int J Syst Bacteriol 49:1631–1643. https://doi.org/10.1099/00207713-49-4-1631
Koren S, Walenz BP, Berlin K et al (2017) Canu: scalable and accurate long-read assembly via adaptive k-mer weighting and repeat separation. Genome Res 27:722–736. https://doi.org/10.1101/gr.215087.116
Kumar S, Stecher G, Li M et al (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. https://doi.org/10.1093/molbev/msy096
Lazzarini A, Cavaletti L, Toppo G, Marinelli F (2000) Rare genera of actinomycetes as potential producers of new antibiotics. Antonie Van Leeuwenhoek 78:399–405
Lee M-J, Kong D, Han K et al (2012) Structural analysis and biosynthetic engineering of a solubility-improved and less-hemolytic nystatin-like polyene in Pseudonocardia autotrophica. Appl Microbiol Biotechnol 95:157–168. https://doi.org/10.1007/s00253-012-3955-x
Li S, Tian X, Niu S et al (2011) Pseudonocardians A–C, new diazaanthraquinone derivatives from a deap-sea actinomycete Pseudonocardia sp. SCSIO 01299. Mar Drugs 9:1428–1439. https://doi.org/10.3390/md9081428
Loveland-Curtze J, Miteva VI, Brenchley JE (2011) NOTE/NOTE Evaluation of a new fluorimetric DNA–DNA hybridization method. Can J Microbiol 255:250–255. https://doi.org/10.1139/W10-121
Maskey RP, Kock I, Helmke E, Laatsch H (2003) Isolation and structure determination of phenazostatin D, a new phenazine froma marine actinomycete isolate Pseudonocardia sp. B6273. Zeitschrift für Naturforsch B 58:692–694. https://doi.org/10.1515/znb-2003-0714
Meier-Kolthoff JP, Auch AF, Klenk HP, Göker M (2013) Genome sequence-based species delimitation with confidence intervals and improved distance functions. BMC Bioinform. https://doi.org/10.1186/1471-2105-14-60
Meier-Kolthoff JP, Göker M (2019) TYGS is an automated high-throughput platform for state-of-the-art genome-based taxonomy. Nat Commun 10:2182. https://doi.org/10.1038/s41467-019-10210-3
Min KL, Hah YC, Lee WY et al (2001) Pseudonocardia kongjuensis sp. nov., isolated from a gold mine cave. Int J Syst Evol Microbiol 51:1505–1510. https://doi.org/10.1099/00207713-51-4-1505
Minnikin DE, Collins MD, Goodfellow M (1979) Fatty acid and polar lipid composition in the classification of cellulomonas, oerskovia and related taxa. J Appl Bacteriol 47:87–95. https://doi.org/10.1111/j.1365-2672.1979.tb01172.x
Moloudizargari M, Mikaili P, Aghajanshakeri S et al (2013) Pharmacological and therapeutic effects of Peganum harmala and its main alkaloids. Pharmacogn Rev 7:199. https://doi.org/10.4103/0973-7847.120524
Oh D-C, Poulsen M, Currie CR, Clardy J (2009) Dentigerumycin: a bacterial mediator of an ant-fungus symbiosis. Nat Chem Biol 5:391–393. https://doi.org/10.1038/nchembio.159
Ramirez-Paredes JG, Larsson P, Thompson KD et al (2020) Reclassification of Francisella noatunensis subsp. orientalis Ottem et al. 2009 as Francisella orientalis sp. nov., Francisella noatunensis subsp. chilensis subsp. nov. and emended description of Francisella noatunensis. Int J Syst Evol Microbiol 70:2034–2048. https://doi.org/10.1099/ijsem.0.004009
Richter M, Rosselló-Móra R (2009) Shifting the genomic gold standard for the prokaryotic species definition. Proc Natl Acad Sci 106:19126–19131. https://doi.org/10.1073/pnas.0906412106
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. https://doi.org/10.1093/oxfordjournals.molbev.a040454
Sasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids. USFCC Newsl 20:1–6
Shin HJ, Lee H-S, Lee D-S (2010) The synergistic antibacterial activity of 1-acetyl-beta-carboline and beta-lactams against methicillin-resistant Staphylococcus aureus (MRSA). J Microbiol Biotechnol 20:501–505. https://doi.org/10.4014/jmb.0910.10019
Shirling EB, Gottlieb D (1966) Methods for characterization of Streptomyces species. Int J Syst Bacteriol 16:313–340. https://doi.org/10.1099/00207713-16-3-313
Staneck JL, Roberts GD (1974) Simplified approach to identification of aerobic actinomycetes by thin-layer chromatography. Appl Microbiol 28:226–231
Sun B, Morikawa T, Matsuda H et al (2004) Structures of new beta-carboline-type alkaloids with antiallergic effects from Stellaria dichotoma(1,2). J Nat Prod 67:1464–1469. https://doi.org/10.1021/np040080a
Taira Z, Kanzawa S, Dohara C et al (1997) Intercalation of Six.BETA.-Carboline Derivatives into DNA. Eisei kagaku 43:83–91. https://doi.org/10.1248/jhs1956.43.83
Tatusova T, Dicuccio M, Badretdin A et al (2016) NCBI prokaryotic genome annotation pipeline. Nucleic Acids Res. https://doi.org/10.1093/nar/gkw569
Thawai C (2018) Pseudonocardia soli sp. nov., isolated from mountain soil. Int J Syst Evol Microbiol 68:1307–1312. https://doi.org/10.1099/ijsem.0.002672
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. https://doi.org/10.1093/nar/22.22.4673
Tian X-P, Long L-J, Li S-M et al (2013) Pseudonocardia antitumoralis sp. nov., a deoxynyboquinone-producing actinomycete isolated from a deep-sea sediment. Int J Syst Evol Microbiol 63:893–899. https://doi.org/10.1099/ijs.0.037135-0
Uezono T, Maruyama W, Matsubara K et al (2001) Norharman, an indoleamine-derived β-carboline, but not Trp-P-2, a γ-carboline, induces apoptotic cell death in human neuroblastoma SH-SY5Y cells. J Neural Transm 108:943–953. https://doi.org/10.1007/s007020170014
Wang W, Nam SJ, Lee BC, Kang H (2008) β-carboline alkaloids from a Korean tunicate Eudistoma sp. J Nat Prod 71:163–166. https://doi.org/10.1021/np070064o
Xing K, Qin S, Bian GK et al (2012) Pseudonocardia nantongensis sp. nov., a novel endophytic actinomycete isolated from the coastal halophyte Tamarix chinensis Lour. Antonie van Leeuwenhoek. Int J Gen Mol Microbiol 102:659–667. https://doi.org/10.1007/s10482-012-9764-5
Yoon SH, Ha S, min, Lim J, et al (2017) A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie van Leeuwenhoek. Int J Gen Mol Microbiol. https://doi.org/10.1007/s10482-017-0844-4
Zhou T-S, Ye W-C, Wang Z-T et al (1998) β-Carboline alkaloids from hypodematium squamuloso-pilosum. Phytochemistry 49:1807–1809. https://doi.org/10.1016/S0031-9422(98)00232-5
Acknowledgements
STP wish to acknowledge University Grants Commission, New Delhi for the Raman Postdoctoral Fellowship for Indian Scholars in the United States of America (F. No. 5-28/2016(IC) dated 10th Feb 2016) and the Director, NCPOR, Ministry of Earth Sciences, Government of India for providing the logistic support for the filed access to Arctic. DM would like to thank Cochin University of Science and Technology and Kerala State Council for Science, Technology and Environment, Govt. of Kerala, India for providing fellowship funding.
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This study was funded by University Grants Commission, New Delhi under the Raman Postdoctoral Fellowship for Indian Scholars in the United States of America (F. No. 5-28/2016(IC) dated 10th Feb 2016) for STP.
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STP, KPK, and JBM conceived and designed research. MD, MU, TSM, SV, RKS and STP conducted experiments. PF, AA, MD and STP analyzed data. MD wrote the manuscript. STP and JBM edited the manuscript. All authors read and approved the manuscript.
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Dhaneesha, M., Umar, M., Merlin, T.S. et al. Pseudonocardia cytotoxica sp. nov., a novel actinomycete isolated from an Arctic fjord with potential to produce cytotoxic compound. Antonie van Leeuwenhoek 114, 23–35 (2021). https://doi.org/10.1007/s10482-020-01490-7
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DOI: https://doi.org/10.1007/s10482-020-01490-7