Production of scopularide A in submerged culture with Scopulariopsis brevicaulis

Abstract

BackgroundMarine organisms produce many novel compounds with useful biological activity, but are currently underexploited. Considerable research has been invested in the study of compounds from marine bacteria, and several groups have now recognised that marine fungi also produce an interesting range of compounds. During product discovery, these compounds are often produced only in non-agitated culture conditions, which are unfortunately not well suited for scaling up. A marine isolate of Scopulariopsis brevicaulis, strain LF580, produces the cyclodepsipeptide scopularide A, which has previously only been produced in non-agitated cultivation.ResultsScopulariopsis brevicaulis LF580 produced scopularide A when grown in batch and fed-batch submerged cultures. Scopularide A was extracted primarily from the biomass, with approximately 7% being extractable from the culture supernatant. By increasing the biomass density of the cultivations, we were able to increase the volumetric production of the cultures, but it was important to avoid nitrogen limitation. Specific production also increased with increasing biomass density, leading to improvements in volumetric production up to 29-fold, compared with previous, non-agitated cultivations. Cell densities up to 36 g L-1 were achieved in 1 to 10 L bioreactors. Production of scopularide A was optimised in complex medium, but was also possible in a completely defined medium.ConclusionsScopularide A production has been transferred from a non-agitated to a stirred tank bioreactor environment with an approximately 6-fold increase in specific and 29-fold increase in volumetric production. Production of scopularide A in stirred tank bioreactors demonstrates that marine fungal compounds can be suitable for scalable production, even with the native production organism.

Effects of Cultural Conditions in Enhancing the Production of Anti-MRSA Activity of Lasiodiplodia pseudotheobromae IBRL OS-64, an Endophytic Fungus Isolated from Leaf of Ocimum sanctum L. in Submerged Fermentation System

“…The present study revealed that the static condition produced the highest anti-MRSA activity and the activity was decreased with the increment of agitation speeds. The result obtained was in agreement with previous studies that reported the highest production of bioactive compound was obtained under static condition (Bringmann et al, 2007;Tamminen et al, 2014). The favorable effect of air flow on the secondary metabolite production could be attributed to the enhancement of product formation by microorganism under-forced aeration.…”

Production of Calcaride A by Calcarisporium sp. in Shaken Flasks and Stirred Bioreactors

“…KF525 generally grew as pellets in flasks and inclusion of agar to obtain filamentous growth ( cf. [ 7 ]) appeared to suppress calcaride A production (data not shown). Therefore, bioreactors were inoculated with pre-cultures containing small pellets and the cultures continued to grow as small pellets (1–3 mm diameter).…”

“…The casamino acid glucose medium described in [ 10 ] does not supply sufficient nitrogen for all of the carbon to be consumed unless carbon is incorporated into storage compounds or secondary metabolites. The carbon–nitrogen ratio (C/N) of this medium was ~67 (g/g), which was much higher than the level at which nitrogen is expected to limit growth (C/N > 10; [ 7 , 20 ]). A high C/N ratio may be useful to trigger production of secondary metabolites such as the calcarides that do not contain nitrogen, and we continued to use medium with high C/N ratios.…”

“…Obtaining adequate growth of the producing organism may also be challenging, particularly for filamentous organisms. However, recently, production of several diverse products from marine fungi has been successfully scaled up in stirred bioreactors (e.g., aspergiolide A [ 5 ], compound 1403C [ 6 ], scopularide A [ 7 ], and pigments [ 8 ]).…”

Effect of Agitation Speed and Cultivation Time on the Production of the Emestrin Produced by Emericella nidulans Marine Fungal

“…Anti-leukemia active compound of sorbicilactone was produced in lab scale from a static cultur of Penicillium chrysogenum (Bringmann et al, 2007). So it was with cytotoxic compound of scopularide that was produced static culture of fungi Scopulariopsis brevicaulis (Tamminen et al, 2014). Our results suggested that, for emestrin production in large scale, it is necessary to improve culture conditions, such as mixing, aeration, and nutrient source optimation.…”