1. Introduction CAS-Croucher Funding Scheme for Joint Laboratories CAS-CF03/04.SC01 CAS-CF03/04.SC01 Isolation and purification of marine bioactive compounds from marine organisms from Sanya Bay Hong Kong University of Science and Technology & South China Sea Institute of Oceanography Marine organisms and their associated microbes are promising sources of antibiotics, antifoulants and marine drugs. In this joint project, our objectives are: 1)To screen for marine bioactive compounds from sponges, gorgonian corals, mangrove plants and their associated microbes from Sanya Bay; 2)To isolate and purify marine bioactive compounds using bioassay-guided separation assay system; and 3)To identify chemical structures of bioactive compounds Sponge Acanthella cavernosa Bacterium P. luteoviolacea Fungus Fusarium sp. Materials & Methods Extract by solvents (I) Screen for bioactive compounds Marine organism Marine microbes Crude extract Test bioactivity Antibacterial 96 well plate Extract Bacteria Inhibition zone Bacteria Extract Petri dish Barnacle TubewormBryozoan Anti-larval settlement 24 well plate Swimming larvae Extract Cytotoxicity Cancer cell lines - MDA-MB-231 - MCF - HeLa 96 well plate Extract (II) Isolation of pure bioactive compounds Active extract Fractionate by HPLC 1 23 4 56 (III) Identification of chemical structures Assay bioactivity of each fraction NMR Further fractionate Pure active compound GC-MSLC-MS 1 HNMR 13 CNMR Culture Research Outcome (I) Bioactive compound screening (III) Bacterial culture collection (IV) Others  Screened >30 sponges, 20 gorgonian corals, 10 mangrove trees, 3 bacteria & 3 fungi  Obtained > 80 compounds, 20 were novel (II) Bioactivity of pure compounds Sesquiterpene Diterpenoids Macrolactin Polyoxygenated steroids  Identified > 20 antifouling, 10 antibacterial & 10 antitumor compounds  Isolated > 600 microbes (130 bacteria, 123 actinomycetes, 27 fungi & others)  Identified 150 bioactive strains  Obtained HPLC profiles for microbial extracts Liquid N 2 storage Liquid N 2 storage -80 o C storage Multiple copies of stocks Antibacterial lactones MIC against E. coli and B. subtilis = 0.1 µg/ml Antilarval diterpenoids Antilarval sesquiterpene Cytotoxic sesquiterpene IC 50 = 8.87 μg/mL EC 50 < 3.2 μg/ml EC 50 = 0.004 – 21.06 μg/ml Antilarval steriods EC 50 = 6.25 – 15.6 μg/ml  Published 24 research articles  Qiu et al. (2006) New polyoxygenated steroids from the South China Sea Gorgonian Echinogorgia aurantiaca. Pharmazie 61:645-647  Qi et al. (2006) Ten new antifouling briarane diterpenoids from the South China Sea gorgonian Junceella juncea. Tetrahedron 62:9123-9130  Lau et al. (2006) Marinomonas ostreistagni sp. nov., isolated from a pearl-oyster culture pond in Sanya, Hainan Province, China. Int J Syst Evol Microbiol 56:2271-2275  Yang et al. (2007) Effect of agitation on violacein production in Pseudoalteromonas luteoviolacea isolated from a marine sponge. Letters in Applied Microbiology,44 (6): 625-630.  Qi et al. (2008) Antifouling and antibacterial compounds of the South China Sea gorgonians Subergorgia suberosa and Scripearia gracillis. Nat Prod Res 22(2): 154-166  Filed 3 US patents, 1 Chinese patent  Trained 2 PhD students & 2 post-docs  Developed further collaboration between the two research teams  Corals and sponges from Sanya Bay are good sources of bioactive compounds; their associated microbes enable better compound supply.  Both physical and chemical culture condition greatly affect compound production and microbial growth. Optimal condition for microbial growth mismatches optimal condition for compound production.  Large culture collection of microbes serves as goldmine of exploration of marine bioactive compounds Conclusion Acknowledgements  Financial support by CAS-Croucher Funding Scheme for Joint Laboratories (CAS-CF03/04.SC01)