BACKGROUND:Oleaginous fungi can accumulate lipids by utilizing a wide range of waste substrates. They are an important source for the industrial production of omega-6 polyunsaturated fatty acids (gamma-linolenic and arachidonic acid) and have been suggested as an alternative route for biodiesel production. Initial research steps for various applications include the screening of fungi in order to find efficient fungal producers with desired fatty acid composition. Traditional cultivation methods (shake flask) and lipid analysis (extraction-gas chromatography) are not applicable for large-scale screening due to their low throughput and time-consuming analysis. Here we present a microcultivation system combined with high-throughput Fourier transform infrared (FTIR) spectroscopy for efficient screening of oleaginous fungi. RESULTS:The microcultivation system enables highly reproducible fungal fermentations throughout 12 days of cultivation. Reproducibility was validated by FTIR and HPLC data. Analysis of FTIR spectral ester carbonyl peaks of fungal biomass offered a reliable high-throughput at-line method to monitor lipid accumulation. Partial least square regression between gas chromatography fatty acid data and corresponding FTIR spectral data was used to set up calibration models for the prediction of saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, unsaturation index, total lipid content and main individual fatty acids. High coefficients of determination (R2 = 0.86-0.96) and satisfactory residual predictive deviation of cross-validation (RPDCV = 2.6-5.1) values demonstrated the goodness of these models. CONCLUSIONS:We have demonstrated in this study, that the presented microcultivation system combined with rapid, high-throughput FTIR spectroscopy is a suitable screening platform for oleaginous fungi. Sample preparation for FTIR measurements can be automated to further increase throughput of the system.

译文

背景:油脂性真菌可以利用多种废物底物来积累脂质。它们是工业生产omega-6多不饱和脂肪酸(γ-亚麻酸和花生四烯酸)的重要来源,并已被建议作为生物柴油生产的替代途径。用于各种应用的初始研究步骤包括筛选真菌,以便找到具有所需脂肪酸组成的有效真菌生产者。传统的培养方法(摇瓶)和脂质分析(萃取气相色谱法)由于通量低且分析费时,因此不适用于大规模筛选。在这里,我们介绍了结合高通量傅立叶变换红外(FTIR)光谱的微培养系统,以有效筛选油性真菌。
结果:微培养系统能够在整个培养的12天中实现高度可重复的真菌发酵。通过FTIR和HPLC数据验证了可重复性。真菌生物质的FTIR光谱酯羰基峰分析提供了一种可靠的高通量在线方法来监测脂质积累。气相色谱脂肪酸数据和相应的FTIR光谱数据之间的偏最小二乘回归用于建立校准模型,用于预测饱和脂肪酸,单不饱和脂肪酸,多不饱和脂肪酸,不饱和指数,总脂质含量和主要单个脂肪酸。较高的测定系数(R2 = 0.86-0.96)和令人满意的交叉验证残差预测偏差(RPDCV = 2.6-5.1)证明了这些模型的优越性。
结论:我们在这项研究中证明,所提出的微培养系统与快速,高通量FTIR光谱相结合是一种适合油性真菌的筛选平台。 FTIR测量的样品制备可以自动化,以进一步提高系统的通量。

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