Direct conversion of straw to ethanol by Fusarium oxysporum: Effect of cellulose crystallinity

Rapid and Effective Oxidative Pretreatment of Woody Biomass at Mild Reaction Conditions and Low Oxidant Loadings

“…Microplate (96-well; Sarstedt, Germany) layout for three-tier screening. Fusarium oxysporum wild-type strain 11C (WT) and putative transformants were inoculated into columns (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12) and screened across five treatments: T1; no ethanol, T2; hygryomcinB (60 μg ml -1 ; Sigma, UK), T3; 0.5% (vv -1 ) ethanol (ethyl alcohol; Sigma, UK), T4; 6% (vv -1 ) ethanol T5; 10% (vv -1 ) ethanol with treatments positioned in rows (A-F) (Primary-tier) (A) or into rows (A-H) and then transferred and Generating and screening Fusarium oxysporum strain 11C transformants. Inoculation of Fusarium oxysporum strain 11C into Mung bean broth [32] for 5 days at 25°C and collection of conidia by filtration through sterile cheesecloth, washing twice with sterile distilled water and adjusting spore concentration to 10 4 conidia per ml (A) Inoculation of Agrobacterium tumefaciens strain AGL-1 to minimal medium (MM) [34] supplemented with Kanamycin (50 μg ml -1 ) for 2 days at 28°C (OD 600nm 0.4-0.6) followed by dilution to OD 600nm of 0.15 in induction medium (IM) [34] supplemented with 200 μM acetosyringone (AS) and incubation for 6 hours at 28°C (B) Co-cultivation of an equal volume of bacterial and fungal cells for 30 minutes at 28°C in liquid co-cultivation medium [34] followed by spreading 100 μl mix onto a UVsterilised cellulose filter membrane placed on solid cocultivation medium on large Petri plates (150x20mm) for 2 days at 25°C where if transformation is successful, T-DNA is transferred from AGL-1 to F. oxysporum strain 11C and randomly integration into the fungal genome results (C) Transfer of cellulose filter membrane to modified selection medium (SM) on large Petri plates (150x20mm) supplemented with 60 hygromycinB for 7-9 days at 25°C followed by isolation of hygromycinB (60 μg ml -1 ) resistant putatively transformed colonies into microtiter (96-well) plates with minimal medium [37] supplemented with hygromycinB (60 μg ml -1 ) for 3 days at 25°C (D) Transfer of putative transformants to microtiter (96-well) plate with minimal medium supplemented with ethanol and butanol selection for primary alcohol tolerance screening followed by purification and PCR prior to two additional rounds of screening (2° and 3°) and selection of candidates for future analysis (E).…”

“…In particular, the broad host range phytopathogen Fusarium oxysporum [11] can degrade and produce ethanol from various cellulosic substrates (e.g. untreated and pre-treated straw [12,13], brewer's spent grain [14], potato waste [15]). Previous work [13] identified F. oxysporum strain 11C as a promising microbial biocatalyst capable of producing high bioethanol yields from delignified wheat straw.…”

Ethanol Production from Cellulose by Two Lignocellulolytic Soil Fungi

“…For ethanol yield calculations, the amount of cellulose in the lignocellulosic substrates used here was considered as 50%. The conversions of cellulose to ethaRol were calculated based on the assumption that 1 g of cellulose on hydrolysis would yield 1.1 g of reducing sugars (25,26).…”

Purification, characterization and mass spectrometric sequencing of transaldolase from Fusarium oxysporum

“…A few microbial species such as Neurospora crassa and Fusarium oxysporum have been reported to acquire the ability of fermenting cellulose directly to ethanol in the 1980s. We have earlier reported on the direct conversion of biomass to ethanol by F. oxysporum [1,2]. Cellulases and xylanases from this microorganism have been characterized [3][4][5][6][7].…”