Optimization of Enniatin Production by Solid-Phase Fermentation of Fusarium tricinctum

Abstract

Enniatins are cyclic depsipeptides produced by fungi of the genus Fusarium that are of interest due to their pronounced biological activities; especially enniatins A, A 1 , B, and B 1 possess anticarcinogenic and anti-HIV properties. In the present study, F. tricinctum was grown on seven solid media and in one liquid growth medium with or without addition of peptone or of amino acid precursors in order to identify favourable media with simple cultivation conditions for maximum enniatin production. Additionally, the optimal duration of growth was investigated for the highest yields of enniatins. From the different media analysed, white beans (Phaseolus vulgaris, solid medium) induced the highest accumulation of enniatins A, A 1 , B, and B 1 , that reached a maximum of 1,365 mg total enniatins in 1 L growth medium after 18 days of fermentation. Fermentation of F. tricinctum on white beans gave the highest yield of enniatins compared to all other media analysed in this study.

Biological Detoxification of Enniatins

“…Initially the main substrates used for enniatin productions were chemically defined liquid media (Audhya and Russell, 1974;Madry et al, 1983;Visconti et al, 1992;, and semi solid media such as corn grit . More recently, several authors have reported the production of high amounts of enniatins using solid matrices such as rice (Moretti et al 2007), corn (Meca et al 2010), or several solid substrates including white beans, soybeans, radish, potato, wheat, corn and rice (Wang et al 2013). According to Wang et al (2013) enniatins production was highest in the white bean media without any supplementation of amino acids (1.365mg/L enniatins A, A1, B1 and B after 18 days).…”

“…Enniatins are strong hydrophobic compounds and their extraction from fungal extracts have been done using mild to strong hydrophobic solvents such as water/methanol and ethyl acetate , methanol , acetonitrile , hexane , chloroform media (Audhya and Russell, 1974;Madry et al, 1983), ethyl acetate (Wang et al, 2013). Among the techniques used so far for the detection and quantification of enniatins from extracts are quantitative paper chromatography or spectrophotometry (Audhya and Russell, 1974;Madry et al, 1983), mass spectrometry/mass spectrometry , HPLC-UV Wang et al, 2013, LC-MS-MS , LC-MS-LIT .…”

“…The fermentation of F. avenaceum DAOM in liquid media resulted in the production of a mixture of enniatins containing predominantly enniatin A, followed by enniatins A1 and B1 (Blais et al, 1991). Wang et al 2013 examined the production of F. tricinctum in seven solid media and in one liquid medium with variation in the supplementation of peptone or amino acid precursors. It was demonstrated that white bean solid media led to the highest enniatins yield in the growing condition chosen, after analysis using HPLC with UV-visible photodiode array detector.…”

“…Solid corn was also used as substrate for enniatin production using F. tricinctum, the separation of the crude extract produced enniatins A, A1, B, B1, B4 and J . Furthermore, extraction of enniatins from crude liquid or solid media has been carried out using non polar solvents such as hexane and dichloromethane (Blais et al, 1991), ethyl acetate (Wang et al, 2013), chloroform (Madry et al, 1983 or moderate non polar solvent mixtures of water and methanol (Cuomo et al, 1991).…”

“…Enniatins are secondary metabolites produced by several Fusarium species such as F. orthocerar (Gäumann et al, 1947) F. tricinctum, F. lateritium, F. oxysporum, F. avenaceum Wang et al, 2013) and a few other genera of fungi such as Alternaria (Mckee et al, 1997), Verticillium (Nilanonta et al, 2003) and Halosarpheia (Lin et al, 2002). More recently, Zobel et al (2005) also demonstrated the production of enniatin B by a recombinant Bacillus subtillis who engineered Aspergillus niger for the production of several variants of enniatin.…”