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.

Comparison of two lipid extraction methods produced by yeast in cheese whey

This work aimed to evaluate nine strains of yeast, previously identified as good producers of lipids in honey medium, for selecting the most suitable strain for the production of lipids in cheese whey medium and compared two well known extraction methods of lipids from the culture medium. The highest yield of total lipids was 1.27 g.L-1 produced by Cryptococcus laurentii 11. A comparison was made between the two culture media: cheese whey and liquid YEPG, and two lipid extraction methods: Bligh and Dyer and Folch et al. for C. laurentii. The experiments were performed with 2² full factorial design using two factors and two levels. Lipid content was higher in cheese whey and there was no difference in the extraction methods statistically. The method of Bligh and Dyer was used in preference to Folch et al. as it resulted in larger mean of total lipids.

Study on Microbial Oil Production with Chlorella pyrenoidosa / 微生物学通报

Chlorella pyrenoidosa No.2 was screened from five species of microalga Chlorella sp. for its higher lipid yield. Effects of medium components and culture conditions on cell growth as well as lipid ac-cumulation of C. pyrenoidosa No.2 were investigated and the results showed that the optimum medium rec-ipe was 20.0 g/L glucose,0.08 g/L glycine,1.0 g/L K2HPO4?3H2O,0.4 g/L MgSO4?7H2O and 0.004 g/L FeSO4?7H2O. The optimum culture temperature,initial pH,shaking rate and light intensity were 28℃,6.0,130 r/min and 650 Lux,respectively. Biomass and lipid content increased from 3.73 g/L and 40.15% to 6.56 g/L and 59.90% when Chlorella pyrenoidosa No.2 was cultivated under the above optimal conditions for 7 days,with lipid yield raised by 162%. Chlorella pyrenoidosa No.2 could produce lipid with xylose as carbon source,and so is potential for lipid production from renewable materials such as lignocellulose. GC analysis demonstrated that the fatty acid composition of the lipid was similar to that of vegetable oil and its unsaturated fatty acid content reached around 71%,thus it is a promising material for biodiesel production.