Tamminen, A.

Wiebe, M.

OceanRep

 Production of bioactive compound(s) with the marine fungus Calcarisporium sp. Anu Tamminen, Yanming Wang and Marilyn Wiebe VTT Technical Research Centre of Finland, P.O. Box 1000, FI-02044, VTT, Finland  Template for VTT scientific posters  Size: A0, ratio 1:1.  You can send a draft of the poster for layout checking to VTT graphic  designers by email:  vtt.visualistit@vtt.fi.   Please remember to leave enough time  for layout checking (3 - 5 days before  the seminar).  Send the poster to printing (pdf or ppt):  esa.tanskanen@vtt.fi  Please attach these instructions for printing: • Dead line (time of delivery) • Delivering address + person (in case you don´t pick up the poster  by yourself from Esa Tanskanen, Tekniikantie 4, Espoo, 1. floor).  Introduction Increasingly, unique bioactive compounds are being obtained from fungi isolated from marine environments. A Calcarisporium sp. (KF525) isolated from the German Wadden Sea has recently been described as producing  interesting cyclic and linear polyesters (calcarides) with antibacterial activity (Silber et al. 2013a) and unrelated, unique calcaripeptides (Silber et al. 2013b). Calcarisporium sp. KF525 is a slow-growing fungus (µmax ~ 0.06 h‒1) and the calcarides are produced during the later stages of growth (or during stationary phase). Large amounts may accumulate in cultures allowed to stand for months, but this is not practical as a sustainable production method. Therefore, production of calcaride A was assessed in flask and bioreactor cultures (Fig. 1). These studies provide a basis for developing an improved production process, which could provide sufficient material for further bioactivity testing. Results  Calcarisporium sp. KF525 grew well on a variety of carbon sources.  Calcaride A was produced well on sucrose, malt extract and starch, substrates which provide slow release of glucose (Fig. 2).  Production was reduced or delayed on glucose and fructose (Fig. 2).  Less calcaride A was produced from fructose than from sucrose in bioreactor cultures, and sucrose was used as carbon source in further experiments.  Calcarisporium sp. KF525 produced 0.8 ± 0.1 mg calcaride A [g biomass]-1 in sucrose containing medium with C/N ~23.  References 1. Silber J, Ohlendorf B, Labes A, Erhard A, Imhoff JF. 2013a. Calcarides A-E, antibacterial macrocyclic and linear polyesters from a Calcarisporium strain. Mar Drugs 11, 3309-3323.  2. Silber J, Ohlendorf B, Labes A, Näther C, Imhoff JF. 2013b. Calcaripeptides A-C, cyclodepsipeptides from a Calcarisporium strain. J. Nat. Prod . 76, 1461-1467. 3. Vogel HJ. 1956. Convenient growth medium for Neurospora (Medium N). Microb Genet Bull 243, 112-119. ACKNOWLEDGMENTS This study was financially supported by the European Commission through the Seventh Framework Programme (FP7-KBBE-2010-4) under grant agreement N° FP7-265926 MARINE FUNGI and is coordinated by Johannes F. Imhoff (KiWiZ, Kieler Wirkstoff-Zentrum am GEOMAR, Am Kiel-Kanal 44, 24106 Kiel, alabes@geomar.de, www.kiwiz.org). Fig. 1. Calcarisporium sp. KF525 (spores and mycelia, left) was grown in 1 L Biostat Q bioreactor (right), inoculated with 10% (v/v) pre-cultures as small pellets.  Fig. 4. Calcaride A production by Calcarisporium sp. KF525 in bioreactors, in defined medium at pH 5.3, with 4% sucrose as carbon source, 3% marine salt, 22°C. The C/N 37 culture was inoculated with more biomass than the other cultures. Results  Calcarisporium sp. KF525 produced calcarides in stirred tank reactors, but specific production was generally lower than in flask cultures.  Calcaride A production was reduced in the presence of excess nitrogen (Fig. 4).   Maximum calcaride A production was obtained in < 14 days, depending on conditions. Fig. 2. Calcaride A and biomass production by Calcarisporium sp. KF525 in defined medium,  flask cultures, pH ~5.3, C/N ~23, with 3% marine salt, 22 C. 2345670 5 10 15 20pHTime (h)pH 6.5pH 5pH 4Fig. 3. Calcaride A and biomass production by Calcarisporium sp. KF525 in casamino acid medium,  flask cultures at various pH values, with 4% sucrose as carbon source, C/N ~66,  3% marine salt, 22°C. 012344 5 6.5Calcaride A (mg [g biomass]-1)pH10 d21 d024684 5 6.5Biomass (g L-1)pH10 d21 d0510152025304 5 6.5Calcaride A (mg L-1)pH10 d21 d0246810Calcaride A (mg L-1)17 d25 d024681012Biomass (g L-1)17 d25 dResults  Calcaride A production was dependent on culture pH (Fig. 3).  Production in cultures initially at pH 6.5 increased after the pH was reduced during growth (Fig. 3), but remained low in bioreactor cultures. 00.10.20.30.40.50.60.70 5 10 15Calcaride A (mg [g biomass]-1)Time (days)C/N 37C/N 24CN/ 170123456789100 5 10 15Calcaride A (mg L-1)Time (days)C/N 37C/N 24CN/ 17Conclusion • Calcaride A is produced by Calcarisporium sp. KF525 at acidic pH values in N-limited cultures.  • Production was partially inhibited in the presence of glucose.  • The time of maximum production was reduced from >3 weeks to < 2 weeks by ensuring optimal conditions for production. • Production of calcaride A has been transferred from flasks to bioreactors.  

Production of bioactive compound(s) with the marine fungus Calcarisporium sp.

Abstract

Similar works

Full text

Available Versions

OceanRep