Single cell protein is a biotechnological product made from microorganisms grown under suitable condition for biomass production. A limited number of food products based on single cell protein is available in global market and the most noted example in terms of tangible profits is the mycoprotein Quorn, produced by Marlow Foods from Fusarium venenatum mycelium. Fungi, in general, are good raw material for food protein production once they grow in a diversity of affordable sustainable substrates as agricultural straw and bagasse, yielding a nutritionally complete biomass along with other metabolites. Therefore, fungi are important cell factories for food industry. Brazil is one of the world’s most biodiverse countries regarding to plants, animals and also fungi, from which innumerable new biotechnological applications can be developed. In the present work Penicillium sclerotiorum was cultivated under varied glucose, peptone and sodium chloride concentrations. Biomass yields ranged between 1.89 g and 7.80 g, being greater in culture media with higher glucose contents. Ash levels ranged from 3.55% to 18.11%, on a dry matter basis (d.m.), with macro and micronutrients ranging from 11.9 to 137.5 mg/100 g for Ca, 87.2 to 154.2 mg/100 g for Mg, 0.9 to 3.0 mg/100 g for Zn (no significant difference), 0.6 to 1.7 mg/100 g for Fe (no significant difference), based on d.m. Protein levels ranged from 21.56% to 41.88% based on d.m. Fatty acid profiles revealed the presence of essential fatty acids linoleic and linolenic, the former in greater amounts in all samples. There were not detected ochratoxins A and B, cyclopiazonic acid, penicillic acid, citrinin and patulin in the analyzed fungal extracts. Besides, as biomass yield and nutritional content were successfully modulated by changing fungal culture media composition, the process showed good potential for industrial development. These results show P. sclerotiorum as a singular producer of nutritious and safe biomass from which a novel single cell protein food can be developed.