Microscale direct transesterification of microbial biomass with ethanol for screening of microorganisms by its fatty acid content

Abstract We present an improved method of direct transesterification suitable for the quantitative analysis of multiple dry samples for its fatty acid content, using a minimal amount of biomass and reactants. The method features an acid-catalyzed direct alcoholysis of microgram samples of dry biomass; the rationale behind the solvent and reagent proportions chosen is discussed. The method was validated using seven microbial strains with diverse lipid content (Saccharomyces cerevisiae, Saccharomyces boulardii, Candida tropicalis, Haematococcus pluvialis, Chlorella vulgaris, Spirulina platensis and Schizochytrium limacinum), and compared with a macroscale direct transesterification method, and with gravimetric analysis of lipids extracted with solvents. The microscale method showed a conversion of 98.06 ± 0.87% of the lipids, using approximately 3 mg of dry biomass, 1mL of 0.2M H2SO4 dissolved in anhydrous ethanol (the acid is the catalyzer and ethanol the reactant)). The mixture was maintained at 70 °C for 20 h with periodic mixing, and then extracted with 2mL n-heptane and analyzed by GC-FID. The lipid content was then calculated considering dilution and sample mass. This method is effective, reliable, and technically attractive for analytical and comparative purposes.

Isolation and screening of microorganisms with potential for biotransformation of terpenic substrates

The objective of the present work was to isolate and select strains with potential to perform the biotransformation of terpenic substrates. Microorganisms obtained from a collection culture and also isolated from a natural source of terpene substrate were tested. Seventeen strains were selected by their resistance to terpenes in potato dextrose agar containing up to 1 percent of limonene or α-pinene and β-pinene (1:1). Subsequently, 10 strains were selected by their capacity of using these terpenes as sole carbon source in a mineral medium. The biotransformation capacity of these strains was tested and the products obtained were identified by GC-MS.

Lab-Scale production of Bacillus atrophaeus' spores by solid state fermentation in fifferent types of bioreactors

Studies were conducted to evaluate Bacillus atrophaeus spores' production by solid-state fermentation (SSF) using sugarcane bagasse as support and soybean molasses as substrate at lab-scale in column bioreactors (forced aeration), plastic bags and Erlenmeyer flasks (aeration by diffusion). Different moisture contents (84 percent, 86 percent and 88 percent; 89 percent, 91 percent and 93 percent) and aeration rates (30mL/min, 45mL/min, 60mL/min and 90mL/min) were studied. The best condition for spore production (3.3x10(10) CFU.g-¹dry matter) in column bioreactor was 80 percent of initial humidity and no aeration. In Erlenmeyer flasks and plastic bags the best sporulation production reached 1.7 up to 4.7x10(10) CFU.g-1dry matter with 88-93 percent of initial moisture. The aeration rate had no significant effect on the spore yield. The initial moisture had a significant effect depending on the bioreactor type. Sporulation kinetic's assay was carried out and it showed the possibility to reduce the time of spore formation in two days.

Estudos foram conduzidos para avaliar a produção de esporos de Bacillus atrophaeus, em escala laboratorial, por fermentação em estado sólido (FES) em biorreatores de coluna (aeração forçada), sacos plásticos e frascos tipo Erlenmeyer (aeração por difusão), usando bagaço de cana como suporte e melaço de soja como substrato. Diferentes teores de umidade (84 por cento, 86 por cento e 88 por cento, 89 por cento, 91 por cento e 93 por cento) e taxas de aeração (30mL/min, 45mL/min, 60mL/min e 90mL/min) foram estudados. A melhor condição para a produção de esporos no biorreator de coluna (3.3 x 10(10) CFU.g-1 matéria seca) foi 80 por cento de umidade inicial, sem aeração. Em frascos Erlenmeyer e sacos de plástico a melhor esporulação foi na faixa de 1.7 a 4.7 x 10(10) CFU.g-1 matéria seca, com 88-93 por cento de umidade inicial. A taxa de aeração não teve efeito significativo sobre o rendimento da esporulação. A umidade inicial apresentou efeito significativo relacionado ao tipo do biorreator. O estudo da cinética da esporulação demonstrou a possibilidade de reduzir o tempo de incubação para esporulação em dois dias.

Coffee residues as substrates for aroma production by Ceratocystis fimbriata in solid state fermentation

Neste trabalho duas diferentes cepas de Ceratocystis fimbriata foram testadas para a producão de aromas frutais em fermentacão no estado sólido (FES) utilizando como substratos casca e polpa de café, suplementados com glicose. Os experimentos foram realizados em frascos Erlenmeyer de 250 mL. As condicões experimentais foram: umidade inicial de 70%, adicão de 20% de glicose e pH 6,0. Os frascos foram cobertos com gaze e a aeracão ocorreu por difusão passiva. A análise do headspace da cultura foi feita por cromatografia gasosa e 12 compostos foram detectados utilizando a casca de café. A análise respirométrica foi realizada para o acompanhamento do crescimento do microrganismo pela determinacão do dióxido de carbono produzido. A producão de ésteres caracterizou o aroma frutal da cultura. A concentracão máxima de voláteis totais foi alcancada após 72 h de cultivo em casca de café (28 µmol.L-1.g-1). Os principais compostos produzidos foram acetato de etila, etanol e acetaldeído, representando 84,7%, 7,6% and 2,0% dos voláteis totais, respectivamente.