Applied Microbiology and Biotechnology

, Volume 72, Issue 5, pp 1063–1073 | Cite as

Effect of culture conditions on mycelial growth, antibacterial activity, and metabolite profiles of the marine-derived fungus Arthrinium c.f. saccharicola

Applied Microbial and Cell Physiology


The effects of culture conditions and competitive cultivation with bacteria on mycelial growth, metabolite profile, and antibacterial activity of the marine-derived fungus Arthrinium c.f. saccharicola were investigated. The fungus grew faster at 30°C, at pH 6.5 and in freshwater medium, while exhibited higher antibacterial activity at 25°C, at pH 4.5, 5.5, and 7.5, and in 34 ppt seawater medium. The fungus grew faster in a high-nitrogen medium that contained 0.5% peptone and/or 0.5% yeast extract, while exhibiting higher bioactivity in a high-carbon medium that contained 2% glucose. The fungal growth was inhibited when it was co-cultured with six bacterial species, particularly the bacterium Pseudoalteromonas piscida. The addition of a cell free culture broth of this bacterium significantly increased the bioactivity of the fungus. Metabolite profiles of the fungus revealed by gas chromatography (GC)-mass spectrometry showed clear difference among different treatments, and the change of relative area of three peaks in GC profile followed a similar trend with the bioactivity variation of fungal extracts. Our results showed clear differences in the optimal conditions for achieving maximal mycelial growth and bioactivity of the fungus, which is important for the further study on the mass cultivation and bioactive compounds isolation from this fungus.



The authors would like to thank Dr. SCK Lau and all the lab mates for their constructive comments in revising the manuscript, and Dr. Virginia Unkefer for proofreading the manuscript. This work was supported by a CAG grant (CA04/05.Sc01) of the Research Grant Council of HKSAR and a CAS-Croucher Foundation grant (CAS-CF03/04.SC01) to PY Qian.


  1. Acar JF (1980) The disc susceptibility test. In: Lorian V (ed) Antibiotics in laboratory medicine, Williams and Wilkins, Baltimore, pp 24–54Google Scholar
  2. Audhya TK, Russell DW (1974) Production of enniatins by Fusarium sambucinum: selection of high-yield conditions from liquid surface cultures. J Gen Microbiol 82:181–190Google Scholar
  3. Banerjee R, De KB, Bhattacharyya BC (1992) Optimization of extracellular protease biosynthesis by a newly isolated Rhizopus oryzae. Indian J Technol 30:275–280Google Scholar
  4. Bode HB, Bethe B, Hofs R, Zeeck A (2002) Big effects from small changes: possible ways to explore nature’s chemical diversity. ChemBioChem 3:619–627CrossRefGoogle Scholar
  5. Bruckner B, Blechschmidt D (1991) Nitrogen regulation of gibberellin biosynthesis in Gibberella fujikuroi. Appl Microbiol Biotechnol 35:646–650Google Scholar
  6. Bugni TS, Ireland CM (2004) Marine-derived fungi: a chemically and biologically diverse group of microorganisms. Nat Prod Rep 21:143–163CrossRefGoogle Scholar
  7. Burgess JG, Jordan EM, Bregu M, Mearns-Spragg AM, Boyd KG (1999) Microbial antagonism: a neglected avenue of natural products research. J Biotechnol 70:27–32CrossRefGoogle Scholar
  8. Calvo AM, Wilson RA, Bok JW, Keller NP (2002) Relationship between secondary metabolism and fungal development. Microbiol Mol Biol Rev 66:447–459CrossRefGoogle Scholar
  9. Candau R, Avalos J, Cerda Olmedo E (1992) Regulation of gibberellin biosynthesis in Gibberella fujikuroi. Plant Physiol 100:1184–1188CrossRefGoogle Scholar
  10. Chisti Y, Moo-Young M (1993) Clean-in-place systems bioreactor: design, validation and operation. ASME Bioeng Div Publ Bed 27:5–12Google Scholar
  11. Cueto M, Jensen PR, Kauffman C, Fenical W, Lobkovsky E, Clardy J (2001) Pestalone, a new antibiotic produced by a marine fungus in response to bacterial challenge. J Nat Prod 64:1444–1446CrossRefGoogle Scholar
  12. Frisvad JC, Samson RA (1991) Filamentous fungi in foods and feeds—ecology, spoilage, and mycotoxin production. In: Arora DK, Mukerji KG, Marth EH (eds) Handbook of applied mycology. Marcel Dekker, New York, pp 31–68Google Scholar
  13. Gallo M, Katz E (1972) Regulation of secondary metabolite biosynthesis: catabolite repression of phenoxazinone synthase and actinomycin formation by glucose. J Bacteriol 109:659–667Google Scholar
  14. Garbayo J, Vilchez C, Nava-Saucedo JE, Barbotin JN (2003) Nitrogen, carbon and light-mediated regulation studies of carotenoid biosynthesis in immobilized mycelia of Gibberella fujikuroi. Enzyme Microb Technol 33:629–634CrossRefGoogle Scholar
  15. Grabley S, Thiericke R (1999) Drug discovery from nature. Grabley S, Thiericke R (eds) Springer, Berlin Heidelberg New York, p 38–48Google Scholar
  16. Hays A, Hobbs G, Smith CP, Oliver SG, Butler PR (1997) Environmental signals triggering methylenomycin production by Streptomyces coelicolor A3 (2). J Bacteriol 179:5511–5515Google Scholar
  17. Lau SCK, Mak KKW, Chen F, Qian PY (2002) Bioactivity of bacterial strains from marine biofilms in Hong Kong waters for the induction of larval settlement in the marine polychaete Hydroides elegans. Mar Ecol Prog Ser 226:301–310Google Scholar
  18. Lau SCK, Tsoi MMY, Li XC, Plakhotnikova I, Wu M, Wong PK, Qian PY (2004) Loktanella hongkongensis sp. nov., a novel member of the α-Proteobacteria originating from marine biofilms in Hong Kong waters. Int J Syst Evol Microbiol 54:2281–2284CrossRefGoogle Scholar
  19. Lau SCK, Tsoi MMY, Li XC, Dobretsov S, Plakhotnikova I, Wu M, Wong PK, Qian PY (2005) Pseudoalteromonas spongiae sp. nov., a new member of the ã -subclass of Proteobacteria isolated from the sponge Mycale adhaerens in Hong Kong waters. Int J Syst Evol Microbiol 55:1593–1596CrossRefGoogle Scholar
  20. Lee OO, Qian PY (2003) Chemical control of bacterial epibiosis and larval settlement of Hydroides elegans in the red sponge Mycale adherens. Biofouling 19:171–180 (suppl)CrossRefGoogle Scholar
  21. Llorens A, Matco R, Hinojo MJ, Logrieco A, Jimenez M (2004) Influence of the interactions among ecological variables in the characterization of Zearalenone producing isolates of Fusarium spp. Syst Appl Microbiol 27:253–260CrossRefGoogle Scholar
  22. Maier A, Maul C, Zerlin M, Grabley S, Thiericke R (1999) Biomolecular-chemical screening a novel screening approach for the discovery of biologically active secondary metabolites II. Application studies with pure metabolites. J Antibiot 52:945–951Google Scholar
  23. Masuma R, Tanaka Y, Omura S (1983) Ammonium ion-depressed fermentation of tylosin by the use of a natural zeolite and its significance in the study of biosynthetic regulation of the antibiotic. J Ferment Technol 61:607–614Google Scholar
  24. Masuma R, Yamaguchi Y, Noumi M, Omura S, Michio N (2001) Effect of seawater concentration on hyphal growth and antimicrobial metabolite production in marine fungi. Mycoscience 42:455–459CrossRefGoogle Scholar
  25. Mearns-Spragg A, Boyd KG, Hubble MO, Burgess JG (1997) Antibiotics from surface-associated marine bacteria. Proceedings of the fourth underwater science symposium. Society for Underwater Technology, London, pp 147–157Google Scholar
  26. Mearns-Spragg A, Bregu M, Boyd KG, Burgess JG (1998) Cross-species induction and enhancement of antimicrobial activity produced by epibiotic bacteria from marine algae and invertebrates, after exposure to terrestrial bacteria. Lett Appl Microbiol 27:142–146CrossRefGoogle Scholar
  27. Miao L, Qian PY (2005) Antagonistic antimicrobial activity of marine fungi and bacteria isolated from natural marine biofilm from Hong Kong waters. Aquat Microb Ecol 38:231–238Google Scholar
  28. Patterson GL, Bolis CM (1997) Fungal cell-wall polysaccharides elicit an antifungal secondary metabolite (phytoalexin) in the cyanobacterium Scytonema acellatum. J Phycol 33:54–60CrossRefGoogle Scholar
  29. Singh CJ (2002) Optimization of an extracellular protease of Chrysosporium keratinophilum and its potential in bioremediation of keratinic wastes. Mycopathologia 156:151–156CrossRefGoogle Scholar
  30. Stevens F (1917) In: Johnston and Stevenson, J Dept Agric P R 1:223Google Scholar
  31. Wang GYS, Abrell LM, Avelar A, Borgeson BM, Crews P (1998) New hirsutane based sesquiterpenes from salt water cultures of a marine sponge-derived fungus and the terrestrial fungus Coriolus consors. Tetrahedron 54:7335–7342CrossRefGoogle Scholar
  32. Yakoby N, Kobiler I, Dinoor A, Prusky D (2000) pH regulation of pectate lyase secretion modulates the attack of Colletotrichum gloeosporioides on avocado fruits. Appl Environ Microbiol 66:1026–1030CrossRefGoogle Scholar
  33. Zhang J, Marcin C, Shifflett MA, Brix T, Salmon P, Greasham R, Buckland B, Chartrain M (1996) Development of a defined medium fermentation process for physostigimine production by Streptomyces grisiofuscus. Appl Microbiol Biotechnol 44:568–575CrossRefGoogle Scholar
  34. Zientz E, Dandekar T, Gross R (2004) Metabolic interdependence of obligate intracellular bacteria and their insect hosts. Microbiol Mol Biol Rev 68:745–770CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Coastal Marine Laboratory, Department of BiologyHong Kong University of Science and TechnologyKowloonChina

Personalised recommendations