The strain Actinosynnema pretiosum FIM06-0063 from Fujian Institute of Microbiology was cultivated on solid state ISP4 media and incubated in 1.5% soluble starch, 0.5% glucose, 1% soybean flour, 0.5% peptone, 0.1% NaCl, 0.5% yeast extract and 0.5% CaCO₃, pH 7.0 medium at 28 ℃ for 24 hours on a rotary shaker (250 r/min ). In the progress of fermention, 6 mL of the vegetative culture were transferred to 250 mL flasks containing 60 mL production medium (1% dextrin, 3% glycerol, 2% soybean flour, 1% corn steep liquor, 0.05% KH₂PO₄, 0.05% NaCl and 0.5% CaCO₃ pH = 7.0) and incubated at 28 ℃ for 5 days on a rotary shaker (250 r/min ).

Jishengella endophytica 161111 was isolated and identified from the healthy roots of the mangrove plant, Xylocarpus granatum, collected from the mangrove reserve zone in Hainan Province, China. The spores of J. endophytica 161111 were directly cultured in 1000 mL Erlenmeyer flasks containing 200 mL fermentation media consisted of glucose 2%, yeast extract 0.5%, peptone 0.5%, KNO₃ 1.5%, CaCO₃ 0.4%, and NaCl 0.4% (pH 7.2). The cultures were incubated on a rotatory shaker at 220 rpm at 28 ℃ for 30 days.

The producing organism, strain TP-A0451 was isolated from a stem of bracken, Pteridium aquilinum, collected in Toyama, Japan. The seed culture was incubated in a medium consisting of 1% soluble starch, 0.5% glucose, 0.3% NZ-case, 0.2% yeast extract, 0.5% tryptone, 0.1% K₂HPO₄, 0.05% MgSO₄.7H₂O and 0.3% CaCO₃(adjusted to pH 7.0 before sterilization) at 30 ℃ for 4 days on a rotary shaker (200rpm). Three-ml aliquots of the seed culture were transferred into thirty 500-ml/K flasks each containing 100 ml of the production medium consisting of 0.5% glucose, 2% glycerol, 2% soluble starch, 1.5% Pharmamedia, 0.3% yeast extract, 1% Diaion HP-20 (adjusted to pH 7.0 before sterilization). Fermentation was carried out for 6 days at 30 ℃ on a rotary shaker (200rpm).

Strains 'MaB-QuH-8' were either derived from an M. aquifolia plant (MaB-QuH-8) from a collection of plant material in Brazil. Agar slant cultures on oatmeal agar (g/l: oatmeal 20, agar 20; pH 7.0-7.2) were used to propagate an inoculum culture on a medium composed as follows (g/l): D-glucose 15, soy flour 15, CaCO₃ 1; KH₂PO₄ 3, NaCl 5;pH 6.5. Cultivation occurred in 500-ml Erlenmeyer flasks containing 100 ml medium for 72 h at 28 ℃ and 180 rpm on a rotary shaker. A 5-ml aliquot of this starting culture was used to inoculate 500-ml Erlenmeyer flasks containing 100 ml of a medium composed as follows (g/l): D-glucose 10, maltose 20, soytone 2, yeast extract 1, KH₂PO₄ 1, MgSO₄.7H₂O 0.5; pH 5.5 (prior to sterilization). The cultivation was carried out for 96 h at 28 ℃ on rotary shakers (180 rpm).

The producing strain Streptomyces sp. neau-D50 was isolated by moist incubation and desiccation method from healthy soybean root in Harbin, Heilongjiang Province, China. Streptomyces sp. neau-D50 was maintained on the yeast extract-malt extract-soluble starch (YMS) medium containing soluble amylum (10 g), yeast extract (2 g), KNO₃ (1 g) and agar (20 g) in 1 L tap water, pH 7. The seed medium consisted of glucose (20 g), soybean flour (15 g) and yeast autolysate (5.0 g) in 1.L water, pH 7. All the media were sterilised at 121℃ for 20 min. A slant culture was incubated for 6-8 days at 28℃ . A total of 10 mL of sterile water was added tothe slant of the YMS medium. The spores were scraped off and transferred into a sterile tube containing glassbeads, and the resulting spore suspension was then filtered through six layers of a sterile cheesecloth and adjusted to 10⁷ -10⁸ colony-forming units (c.f.u.) m/L using YMS medium. A 250-mL flask containing 20 mL of seed medium was inoculated with the spore suspension (2 mL) and incubated at 28℃ for 24 h, on a shaker operating at 250 r.p.m. Then, 8 mL of the culture was transferred into a 1-L Erlenmeyer flask containing 100 mL of the fermentation medium comprising glucose (0.5%), lactose (1.5%), cotton seed powder (2.0%), FeSO₄.7H₂O (0.6%), MnSO₄.H₂O (0.2%), MgSO₄.7H₂O (0.2%) and CaCO₃ (0.3%), pH 7. Fermentation was carried out at 28℃ for 7 days on a rotary shaker at 250 r.p.m.

The organism Streptomyces sp. (DSM 11575) was isolated from root nodules of Alnus glutinosa collected near Jena (Germany). The fermentation was carried out for 14 days at 28 ℃ in four fermentors containing 5 liters of a medium composed as follows: glucose 1%, Na₂PO₄.2H₂O 0.069%, KH₂PO₄ 0.026%, MgSO₄.7H₂O 0.02%, Na(Fe)EDTA 0.001%, NaCl 0.03%, salt-solution 1 ml [H₃BO₄ 1.5.10^-4%, MnSO₄ 8.10^-5%, ZnSO₄.7H₂O 6.10^-5%, CuSO₄.5H₂O 1.10^-5%, NaMoO₄.2H₂O 2.5.10^-6% and CoSO₄.7H₂O 1.10-7%], pH 6.8.

Healthy specimens of the Egyptian sponge Aplysina fistularis were collected from Sharm El-Sheikh from January to February 2008. Induction of mutation by UV irradiation: Spores of Hedaya48 were gently scraped from the surface of ISP-2 agar plates, washed with sterile normal saline (0.90%) and filtered through glass wool. Spore suspensions were checked microscopically and diluted to have a count of 10⁴ spore/ml. Three milliliters of spore suspension was exposed to UV light (Philips TUV 30-W lamp) for different exposure times (5, 10, 15, 20, 25, 30, 35 and 40 min) placed about 25 cm above the liquid surface and gently swirled in a petri dish. After incubation in the dark, spores were plated on ISP-2 agar, incubated at 28 ℃ and observed after 72 h. Mutation, survival rates and antibiotic production were determined. Optimization of saadamycin production: The optimization of production of the anti-mycotic antibiotic, saadamycin, was carried out in 250-ml Erlenmeyer flasks containing 50 ml of starch nitrate medium and monitored in terms of mcg/ml. Duplicate flasks were pooled for analysis, and each result was an average of triplicate assays. Each parameter optimized earlier was incorporated in subsequent experiments. Optimized medium: Finally, production medium containing (g/l) starch, 10; glucose, 10; NaNO₃,1.0; valine, 0.5; alanine, 0.25; phenylalanine, 0.25; KH₂PO₄, 1.0; MgSO₄, 0.5; CaCO₃, 2.0; NaCl, 1.0; FeSO₄, 0.2; seawater 1 l; and pH 6.5 at 35 ℃ was recommended for saadamycin production by mutant Ah22.

Healthy specimens of the Egyptian sponge Aplysina fistularis were collected from Sharm El-Sheikh from January to February 2008. Induction of mutation by UV irradiation: Spores of Hedaya48 were gently scraped from the surface of ISP-2 agar plates, washed with sterile normal saline (0.90%) and filtered through glass wool. Spore suspensions were checked microscopically and diluted to have a count of 10⁴ spore/ml. Three milliliters of spore suspension was exposed to UV light (Philips TUV 30-W lamp) for different exposure times (5, 10, 15, 20, 25, 30, 35 and 40 min) placed about 25 cm above the liquid surface and gently swirled in a petri dish. After incubation in the dark, spores were plated on ISP-2 agar, incubated at 28 ℃ and observed after 72 h. Mutation, survival rates and antibiotic production were determined. Optimization of saadamycin production: The optimization of production of the anti-mycotic antibiotic, saadamycin, was carried out in 250-ml Erlenmeyer flasks containing 50 ml of starch nitrate medium and monitored in terms of mcg/ml. Duplicate flasks were pooled for analysis, and each result was an average of triplicate assays. Each parameter optimized earlier was incorporated in subsequent experiments. Optimized medium: Finally, production medium containing (g/l) starch, 10; glucose, 10; NaNO₃,1.0; valine, 0.5; alanine, 0.25; phenylalanine, 0.25; KH₂PO₄, 1.0; MgSO₄, 0.5; CaCO₃, 2.0; NaCl, 1.0; FeSO₄, 0.2; seawater 1 l; and pH 6.5 at 35 ℃ was recommended for saadamycin production by mutant Ah22.

Streptomyces sp. GT20021503 was isolated from the stem of B. gymnorrhiza. Liquid organic medium 79 (dextrose 10 g, bacto peptone 10 g, casamino acids 1 g, yeast extract 2 g, NaCl 6 g, H₂O 1 L) (2 × 100 mL/flask) was inoculated with a suspension of mycelium and spores (about 1 × 1 cm) of the title strain grown on agar slants or agar plates (oatmeal 20 g, agar 18 g, H₂O 1 L, pH 7.2,). After incubation for 48 h on a rotary shaker at 28 ℃, the culture was transferred to 3200 mL of medium 2 (oatmeal 20 g, CaCl₂ 5.7 × 10^-3 g, Fe-citrate 2.5 × 10^-3 g, MnSO₄ 5.0 × 10^-4 g, ZnCl₂ 2.5 × 10^-4 g, CuSO₄ 4.0 × 10^-5 g, Na₂B₄O₂ 5.0 × 10^-5g, CoCl 1.0 × 10^-5 g, NaMoO₄ 2.1x 10^-5g, H₂O 1 L, pH 7.8) (eight 1000 mL-scale Erlenmeyer flasks with 400 mL of medium 2 each) and incubated at 28 ℃ under shaking conditions for 48 h to yield prefermentation culture, which was poured into a 300 L-scale fermenter filled with 200 L of medium 2 and fermented for 5 days.

The producing microorganism, strain TP-A0456, was isolated from the wild plant of Cryptomeria japonica collected in Kosugi-machi, Toyama, Japan. Streptomyces sp. TP-A0456 cultured on a slant agar medium was inoculated into five 500-ml K-l flasks each containing 100 ml of the seed medium consisting of soluble starch 1%, glucose 0.5%, NZ-case (Humco Scheffield Chemical Co.) 0.3%, yeast extract (Difco Laboratories) 0.2%, tryptone (Difco Laboratories) 0.5%, K₂HPO₄ 0.1%, MgSO₄-7H₂O 0.05% and CaCO₃ 0.3% (pH 7.0). The inoculated flasks were cultivated on a rotary shaker (200 rpm) at 30℃ for 4days. Three-ml of the seed culture was transferred into a hundred 500-ml K-l flasks each containing 100 ml of the production mediumconsisting of glucose 0.5%, glycerol 2%, soluble starch 2%, Pharmamedia (Traders Protein) 1.5%, yeast extract (Difco Laboratories) 0.3% and Diaion HP-20 (Mitsubishi Chemical Co.) 1%. The pH of the medium was adjusted to 7.0 before sterilization. The inoculated flasks were cultured on a rotary shaker (200 rpm) at 30 ℃ for 6 days.

The producing strain TP-A0595 was isolated from a leaf of leek Allium tuberosum collected in Toyama, Japan. Strain TP-A0595 was cultured at 30 ℃ for 4 days on a rotary shaker (200rpm) in four 500 ml/K flasks each containing 100 ml of the seed medium consisting of soluble starch 1%, glucose 0.5%, NZ-case (Humco Scheffield Chemical Co.) 0.3%, yeast extract (Difco Laboratories) 0.2%, tryptone (Difco Laboratories) 0.5%, K₂HPO₄ 0.1%, MgSO₄.7H₂O 0.05% and CaCO₃ 0.3% (pH 7.0). Three-ml of the seed culture were transferred into one hundred 500 ml/K flasks each containing 100 ml of the production medium consisting of glucose 0.5%, glycerol 2%, soluble starch 2%, Pharmamedia (Difco Laboratories) 1.5%, yeast extract 0.3% and Diaion HP-20 (Mitsubishi Chemical Co.) 1%. The pH of the medium was adjusted to 7.0 before sterilization. The inoculated flasks were shaken on a rotary shaker (200rpm) at 30 ℃ for 6 days.

The endophytic fungus 0248 was isolated from healthy garlic based on antifungal activity. Fresh mycelium were picked and inoculated into 500 mL Erlenmeyer flasks containing 100 mL PD medium. After 2 days of incubation at 28 ± 1 °C on a rotary shaker at 150 rpm, 3 mL culture liquid was transferred as seeds into a 300-mL Erlenmeyer flask containing 30 mL medium (20 gL 1 dextrose, 5 gL 1 peptone, 1 gL 1 beef extract, 0.001 gL 1 ZnSO₄7H₂O and 0.01 gL 1 NH₄Cl). The resulting culture was kept on a rotary shaker at 180 rpm for 4 days at 28 ± 1 °C.

Strain MG-37 was isolated from sediment collected by us (ATB, GK) in the Raune Fjord, Norway (N 60° 15.898, E 5° 08.237) at a depth of 250 meters. Strain MG-37 was cultivated in a 10-liter stirred tank fermentor (Biostat S, B. Braun Melsungen, Germany) in a complex medium that consisted of soluble starch 10 g, glucose 10 g, glycerol 10 g, cornsteep powder (Marcor) 2.5 g, Bacto peptone 5.0 g, yeast extract (Ohly Kat) 2.0 g, NaCl 1.0 g and CaCO₃ 3.0 g in 1.0 liter tap water, adjusted to pH 7.3 prior to sterilization. The fermentor was inoculated with 5 vol-% of shake cultures, grown in 500-ml Erlenmeyer flasks with one baffle for 48 hours on a rotary shaker at 120 rpm and 27 ℃ using the same medium. The fermentation was carried out for 96 hours at 27 ℃ with an aeration rate of 0.5 liter/liter culture/minute and an agitation of 250 rpm.