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Planta Med 2019; 85(06): 503-512
DOI: 10.1055/a-0835-2332
Natural Product Chemistry and Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Induction of Secondary Metabolites from the Marine-Derived Fungus Aspergillus versicolor through Co-cultivation with Bacillus subtilis

Nada M. Abdel-Wahab   *
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
2   Department of Pharmacognosy, Faculty of Pharmacy, Minia University, Minia, Egypt
,
Sebastian Scharf   *
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
,
Ferhat C. Özkaya
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
,
Tibor Kurtán
3   Department of Organic Chemistry, University of Debrecen, Debrecen, Hungary
,
Attila Mándi
3   Department of Organic Chemistry, University of Debrecen, Debrecen, Hungary
,
Mostafa A. Fouad
2   Department of Pharmacognosy, Faculty of Pharmacy, Minia University, Minia, Egypt
,
Mohamed S. Kamel
2   Department of Pharmacognosy, Faculty of Pharmacy, Minia University, Minia, Egypt
4   Department of Pharmacognosy, Faculty of Pharmacy, Deraya University, New Minia, Egypt
,
Werner E. G. Müller
5   Institut für Physiologische Chemie, Universitätsmedizin der Johannes-Gutenberg-Universität Mainz, Mainz, Germany
,
Rainer Kalscheuer
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
,
Wenhan Lin
6   State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, China
,
Georgios Daletos
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
,
Weaam Ebrahim
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
7   Department of Pharmacognosy, Faculty of Pharmacy, Mansoura University, Mansoura, Egypt
,
Zhen Liu
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
,
Peter Proksch
1   Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
› Author Affiliations
Further Information

Publication History

received 24 August 2018
revised 09 January 2019

accepted 13 January 2019

Publication Date:
30 January 2019 (online)

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Abstract

A new cyclic pentapeptide, cotteslosin C (1), a new aflaquinolone, 22-epi-aflaquinolone B (3), and two new anthraquinones (9 and 10), along with thirty known compounds (2, 4 – 8, 11 – 34) were isolated from a co-culture of the sponge-associated fungus Aspergillus versicolor with Bacillus subtilis. The new metabolites were only detected in the co-culture extract, but not when the fungus was grown under axenic conditions. Furthermore, the co-culture extract exhibited an enhanced accumulation of the known constituents versicolorin B (14), averufin (16), and sterigmatocyctin (19) by factors of 1.5, 2.0, and 4.7, respectively, compared to the axenic fungal culture. The structures of the isolated compounds were elucidated on the basis of 1D and 2D NMR spectra and mass spectrometry as well as by comparison with literature data. The absolute configuration of compounds 3, 9, and 10 was determined by ECD (electronic circular dichroism) analysis aided by TDDFT-ECD (time-dependent density functional theory electronic circular dichroism) calculations. Compounds 15, 18 – 21, and 26 exhibited strong to moderate cytotoxic activity against the mouse lymphoma cell line L5178Y, with IC50 values ranging from 2.0 to 21.2 µM, while compounds 14, 16, 31, 32, and 33 displayed moderate inhibitory activities against several gram-positive bacteria, with MIC values ranging from 12.5 to 50 µM.

Key words

sponge-derived fungus - Aspergillus versicolor - cyclic pentapeptide - co-culture - ECD calculations - cytotoxicity - antibacterial activity

* These authors contributed equally to this work.


Supporting Information

    HRESIMS and NMR spectra of compounds 1, 3, 9, and 10, ECD calculations for compound 10, HPLC chromatograms of EtOAc extracts from co-culture experiments and Marfeyʼs analysis for compound 1 are available as Supporting Information.

  • Supporting Information
 

  • References

  • 1 Rateb ME, Ebel R. Secondary metabolites of fungi from marine habitats. Nat Prod Rep 2011; 28: 290-344
    CrossrefPubMedGoogle Scholar
  • 2 Almeida C, El Maddah F, Kehraus S, Schnakenburg G, König GM. Endolides A and B, vasopressin and serotonin-receptor interacting N-methylated peptides from the sponge-derived fungus Stachylidium sp. Org Lett 2016; 18: 528-531
    CrossrefPubMedGoogle Scholar
  • 3 Liu J, Gu B, Yang L, Yang F, Lin H. New anti-inflammatory cyclopeptides from a sponge-derived fungus Aspergillus violaceofuscus . Front Chem 2018; 6: 226
    CrossrefPubMedGoogle Scholar
  • 4 Brakhage AA, Schroeckh V. Fungal secondary metabolites – strategies to activate silent gene clusters. Fungal Genet Biol 2011; 48: 15-22
    CrossrefPubMedGoogle Scholar
  • 5 Marmann A, Aly A, Lin W, Wang B, Proksch P. Co-cultivation – a powerful emerging tool for enhancing the chemical diversity of microorganisms. Mar Drugs 2014; 12: 1043-1065
    CrossrefPubMedGoogle Scholar
  • 6 Liu S, Dai H, Heering C, Janiak C, Lin W, Liu Z, Proksch P. Inducing new secondary metabolites through co-cultivation of the fungus Pestalotiopsis sp. with the bacterium Bacillus subtilis . Tetrahedron Lett 2017; 58: 257-261
    CrossrefPubMedGoogle Scholar
  • 7 Ebrahim W, El-Neketi M, Lewald LI, Orfali RS, Lin W, Rehberg N, Kalscheuer R, Daletos G, Proksch P. Metabolites from the fungal endophyte Aspergillus austroafricanus in axenic culture and in fungal-bacterial mixed cultures. J Nat Prod 2016; 79: 914-922
    CrossrefPubMedGoogle Scholar
  • 8 Abdelwahab MF, Kurtán T, Mándi A, Müller WEG, Fouad MA, Kamel MS, Liu Z, Ebrahim W, Daletos G, Proksch P. Induced secondary metabolites from the endophytic fungus Aspergillus versicolor through bacterial co-culture and OSMAC approaches. Tetrahedron Lett 2018; 59: 2647-2652
    CrossrefPubMedGoogle Scholar
  • 9 Fremlin LJ, Piggott AM, Lacey E, Capon RJ. Cottoquinazoline A and cotteslosins A and B, metabolites from an Australian marine-derived strain of Aspergillus versicolor . J Nat Prod 2009; 72: 666-670
    CrossrefPubMedGoogle Scholar
  • 10 Neff SA, Lee SU, Asami Y, Ahn JS, Oh H, Baltrusaitis J, Gloer JB, Wicklow DT. Aflaquinolones A–G: secondary metabolites from marine and fungicolous isolates of Aspergillus spp. J Nat Prod 2012; 75: 464-472
    CrossrefPubMedGoogle Scholar
  • 11 Hamasaki T, Hatsuda Y, Terashima N, Renbutsu M. The structure of two new metabolites of Aspergillus versicolor . Agric Biol Chem 1965; 29: 166-167
    CrossrefPubMedGoogle Scholar
  • 12 Jakšić D, Puel O, Canlet C, Kopjar N, Kosalec I, Klarić MŠ. Cytotoxicity and genotoxicity of versicolorins and 5-methoxysterigmatocystin in A549 cells. Arch Toxicol 2012; 86: 1583-1591
    CrossrefPubMedGoogle Scholar
  • 13 Mándi A, Mudianta IW, Kurtán T, Garson MJ. Absolute configuration and conformational study of psammaplysins A and B from the balinese marine sponge Aplysinella strongylata . J Nat Prod 2015; 78: 2051-2056
    CrossrefPubMedGoogle Scholar
  • 14 Superchi S, Scafato P, Górecki M, Pescitelli G. Absolute configuration determination by quantum mechanical calculation of chiroptical spectra: basics and applications to fungal metabolites. Curr Med Chem 2017; 25: 287-320
    PubMedGoogle Scholar
  • 15 Castonguay A, Brassard P. C-Alkylation of 1,3-dihydroxyanthraquinones. Total syntheses of (±)-averufin and (±)-bipolarin. Can J Chem 1977; 55: 1324-1332
    CrossrefPubMedGoogle Scholar
  • 16 Hodge RP, Harris CM, Harris TM. Verrucofortine, a major metabolite of Penicillium verrucosum var. cyclopium, the fungus that produces the mycotoxin verrucosidin. J Nat Prod 1988; 51: 66-73
    CrossrefPubMedGoogle Scholar
  • 17 Pan C, Shi Y, Chen X, Chen CTA, Tao X, Wu B. New compounds from a hydrothermal vent crab-associated fungus Aspergillus versicolor XZ-4. Org Biomol Chem 2017; 15: 1155-1163
    CrossrefPubMedGoogle Scholar
  • 18 Lee YM, Li H, Hong J, Cho HY, Bae KS, Kim MA, Kim DK, Jung JH. Bioactive metabolites from the sponge-derived fungus Aspergillus versicolor . Arch Pharmacal Res 2010; 33: 231-235
    CrossrefPubMedGoogle Scholar
  • 19 Steyn PS, Vleggaar R, Wessels PL, Cole RJ. Structure and carbon-13 nuclear magnetic resonance assignments of versiconal acetate, versiconol acetate, and versiconol, metabolites from cultures of Aspergillus parasiticus treated with dichlorvos. J Chem Soc Perkin Trans 1979; 1: 451-459
    PubMedGoogle Scholar
  • 20 Dou Y, Wang X, Jiang D, Wang H, Jiao Y, Lou H, Wang X. Metabolites from Aspergillus versicolor, an endolichenic fungus from the lichen Lobaria retigera . Drug Discoveries Ther 2014; 8: 84-88
    PubMedGoogle Scholar
  • 21 Yamazaki M, Satoh Y, Maebayashi Y, Horie Y. Monoamine oxidase inhibitors from a fungus: Emericella navahoensis . Chem Pharm Bull 1988; 36: 670-675
    CrossrefPubMedGoogle Scholar
  • 22 Kikuchi N, Teshiba M, Tsutsumi T, Fudou R, Nagasawa H, Sakuda S. Endocrocin and its derivatives from the Japanese mealybug Planococcus kraunhiae . Biosci Biotechnol Biochem 2011; 75: 764-767
    CrossrefPubMedGoogle Scholar
  • 23 Hamasaki T, Matsui K, Isono K, Hatsuda Y. A new metabolite from Aspergillus versicolor . Agric Biol Chem 1973; 37: 1769-1770
    CrossrefPubMedGoogle Scholar
  • 24 Kim HS, Park IY, Park YJ, Lee JH, Hong YS, Lee JJ. A novel dihydroxanthenone, AGI-B4 with inhibition of VEGF-induced endothelial cell growth. J Antibiot 2002; 55: 669-672
    CrossrefPubMedGoogle Scholar
  • 25 Hamasaki T, Sato Y, Hatsuda Y. Structure of sydowinin A, sydowinin B, and sydowinol, metabolites from Aspergillus sydowi . Agric Biol Chem 1975; 39: 2341-2345
    PubMedGoogle Scholar
  • 26 Kato H, Yoshida T, Tokue T, Nojiri Y, Hirota H, Ohta T, Williams RM, Tsukamoto S. Notoamides A–D: prenylated indole alkaloids isolated from a marine derived fungus, Aspergillus sp. Angew Chem Int Ed Engl 2007; 119: 2304-2306
    CrossrefPubMedGoogle Scholar
  • 27 Chang YW, Yuan CM, Zhang J, Liu S, Cao P, Hua HM, Di YT, Hao XJ. Speramides A–B, two new prenylated indole alkaloids from the freshwater-derived fungus Aspergillus ochraceus KM007. Tetrahedron Lett 2016; 57: 4952-4955
    CrossrefPubMedGoogle Scholar
  • 28 Tsukamoto S, Kato H, Greshock TJ, Hirota H, Ohta T, Williams RM. Isolation of notoamide E, a key precursor in the biosynthesis of prenylated indole alkaloids in a marine-derived fungus, Aspergillus sp. J Am Chem Soc 2009; 131: 3834-3835
    CrossrefPubMedGoogle Scholar
  • 29 Qian-Cutrone J, Huang S, Shu YZ, Vyas D, Fairchild C, Menendez A, Krampitz K, Dalterio R, Klohr SE, Gao Q. Stephacidin A and B: two structurally novel, selective inhibitors of the testosterone-dependent prostate LNCaP cells. J Am Chem Soc 2002; 124: 14556-14557
    CrossrefPubMedGoogle Scholar
  • 30 Tsukamoto S, Umaoka H, Yoshikawa K, Ikeda T, Hirota H. Notoamide O, a structurally unprecedented prenylated indole alkaloid, and notoamides P–R from a marine-derived fungus, Aspergillus sp. J Nat Prod 2010; 73: 1438-1440
    CrossrefPubMedGoogle Scholar
  • 31 Lee SU, Asami Y, Lee D, Jang JH, Ahn JS, Oh H. Protuboxepins A and B and protubonines A and B from the marine-derived fungus Aspergillus sp. SF-5044. J Nat Prod 2011; 74: 1284-1287
    CrossrefPubMedGoogle Scholar
  • 32 Ishikawa N, Tanaka H, Koyama F, Noguchi H, Wang CC, Hotta K, Watanabe K. Non-Heme dioxygenase catalyzes atypical oxidations of 6,7-bicyclic systems to form the 6,6-quinolone core of viridicatin-type fungal alkaloids. Angew Chem Int Ed Engl 2014; 126: 13094-13098
    CrossrefPubMedGoogle Scholar
  • 33 Trisuwan K, Rukachaisirikul V, Borwornwiriyapan K, Phongpaichit S, Sakayaroj J. Benzopyranone, benzophenone, and xanthone derivatives from the soil fungus Penicillium citrinum PSU-RSPG95. Tetrahedron Lett 2014; 55: 1336-1338
    CrossrefPubMedGoogle Scholar
  • 34 Gao H, Zhou L, Cai S, Zhang G, Zhu T, Gu Q, Li D. Diorcinols B–E, new prenylated diphenyl ethers from the marine-derived fungus Aspergillus versicolor ZLN-60. J Antibiot 2013; 66: 539-542
    CrossrefPubMedGoogle Scholar
  • 35 Li XB, Zhou YH, Zhu RX, Chang WQ, Yuan HQ, Gao W, Zhang LL, Zhao ZT, Lou HX. Identification and biological evaluation of secondary metabolites from the endolichenic fungus Aspergillus versicolor . Chem Biodivers 2015; 12: 575-592
    CrossrefPubMedGoogle Scholar
  • 36 Sassa T, Takemura T, Ikeda M, Miura Y. Structure of radiclonic acid, a new plant growth-regulator produced by a fungus. Tetrahedron Lett 1973; 14: 2333-2334
    CrossrefPubMedGoogle Scholar
  • 37 Gao W, Jiang L, Ge L, Chen M, Geng C, Yang G, Li Q, Ji F, Yan Q, Zou Y, Zhong L, Liu X. Sterigmatocystin-induced oxidative DNA damage in human liver-derived cell line through lysosomal damage. Toxicol In Vitro 2015; 29: 1-7
    CrossrefPubMedGoogle Scholar
  • 38 Liu K, Zheng Y, Miao C, Xiong Z, Xu L, Guan H, Yang Y, Zhao L. The antifungal metabolites obtained from the rhizospheric Aspergillus sp. YIM PH30001 against pathogenic fungi of Panax notoginseng . Nat Prod Res 2014; 28: 2334-2337
    CrossrefPubMedGoogle Scholar
  • 39 Li ZX, Wang XF, Ren GW, Yuan XL, Deng N, Ji GX, Li W, Zhang P. Prenylated diphenyl ethers from the marine algal-derived endophytic fungus Aspergillus tennesseensis . Molecules 2018; 23: E2368
    CrossrefPubMedGoogle Scholar
  • 40 Xu X, Yang H, Xu H, Yin L, Chen Z, Shen H. Diphenyl ethers from a marine-derived isolate of Aspergillus sp. CUGB-F046. Nat Prod Res 2018; 32: 821-825
    CrossrefPubMedGoogle Scholar
  • 41 Kjer J, Debbab A, Aly AH, Proksch P. Methods for isolation of marine-derived endophytic fungi and their bioactive secondary products. Nat Protoc 2010; 5: 479-490
    CrossrefPubMedGoogle Scholar
  • 42 Ashour M, Edrada R, Ebel R, Wray V, Wätjen W, Padmakumar K, Müller WEG, Lin WH, Proksch P. Kahalalide derivatives from the Indian sacoglossan mollusk Elysia grandifolia . J Nat Prod 2006; 69: 1547-1553
    CrossrefPubMedGoogle Scholar
  • 43 Clinical and Laboratory Standards Institute. Methods for Dilution antimicrobial Susceptibility Tests for Bacteria that grow aerobically. Approved Standard – Ninth Ed. CLSI Document M07-A9. Wayne, PA: Clinical and Laboratory Standards Institute; 2012
    Google Scholar
  • 44 MacroModel. New York: Schrödinger LLC; 2015. Available at https://www.schrodinger.com/macromodel Accessed January 23, 2018
    Google Scholar
  • 45 Yanai T, Tew DP, Handy NC. A new hybrid exchange – correlation functional using the Coulomb-attenuating method (CAM-B3LYP). Chem Phys Lett 2004; 393: 51-57
    CrossrefPubMedGoogle Scholar
  • 46 Pescitelli G, Bruhn T. Good computational practice in the assignment of absolute configurations by TDDFT calculations of ECD spectra. Chirality 2016; 28: 466-474
    CrossrefPubMedGoogle Scholar
  • 47 Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA, Cheeseman JR, Scalmani G, Barone V, Mennucci B, Petersson GA, Nakatsuji H, Caricato M, Li X, Hratchian HP, Izmaylov AF, Bloino J, Zheng G, Sonnenberg JL, Hada M, Ehara M, Toyota K, Fukuda R, Hasegawa J, Ishida M, Nakajima T, Honda Y, Kitao O, Nakai H, Vreven T, Montgomery jr. JA, Peralta JE, Ogliaro F, Bearpark M, Heyd JJ, Brothers E, Kudin KN, Staroverov VN, Kobayashi R, Normand J, Raghavachari K, Rendell A, Burant JC, Iyengar SS, Tomasi J, Cossi M, Rega N, Millam JM, Klene M, Knox JE, Cross JB, Bakken V, Adamo C, Jaramillo J, Gomperts R, Stratmann RE, Yazyev O, Austin AJ, Cammi R, Pomelli C, Ochterski JW, Martin RL, Morokuma K, Zakrzewski VG, Voth GA, Salvador P, Dannenberg JJ, Dapprich S, Daniels AD, Farkas Ö, Foresman JB, Ortiz JV, Cioslowski J, Fox DJ. Gaussian 09, Revision B.01. Wallingford, CT: Gaussian, Inc.; 2010
    Google Scholar
  • 48 Stephens PJ, Harada N. ECD cotton effect approximated by the Gaussian curve and other methods. Chirality 2010; 22: 229-233
    PubMedGoogle Scholar
  • 49 Varetto U. MOLEKEL 5.4. Manno, Switzerland: Swiss National Supercomputing Centre; 2009
    Google Scholar