Shapaval Volha, Afseth Nils Kristian, Vogt Gjermund, Kohler Achim
Nofima AS, Centre for Biospectroscopy and Data modeling, Osloveien 1, N-1430 Ås, Norway.
Microb Cell Fact. 2014 Sep 11;13(1):86. doi: 10.1186/1475-2859-13-86.
Fungal production of polyunsaturated fatty acids (PUFAs) is a highly potential approach in biotechnology. Currently the main focus is directed towards screening of hundreds strains in order to select of few potential ones. Thus, a reliable method for screening a high number of strains within a short period of time is needed. Here, we present a novel method for screening of PUFA-producing fungi by high-throughput microcultivation and FTIR spectroscopy. In the study selected Mucor fungi were grown in media with different carbon sources and fatty acid profiles were predicted on the basis of the obtained spectral data. FTIR spectra were calibrated against fatty acid analysis by GC-FD. The calibration models were cross-validated and correlation coefficients (R2) from 0.71 to 0.78 with RMSECV (root mean squared error) from 2.86% to 6.96% (percentage of total fat) were obtained. The FTIR results show a strong correlation to the results obtained by GC analysis, where high total contents of unsaturated fatty acids (both PUFA and MUFA) were achieved for Mucor plumbeus VI02019 cultivated in canola, olive and sunflower oil and Mucor hiemalis VI01993 cultivated in canola and olive oil.
真菌生产多不饱和脂肪酸(PUFAs)是生物技术中一种极具潜力的方法。目前主要集中于筛选数百种菌株,以挑选出少数有潜力的菌株。因此,需要一种可靠的方法在短时间内筛选大量菌株。在此,我们提出一种通过高通量微培养和傅里叶变换红外光谱(FTIR)对产PUFA真菌进行筛选的新方法。在该研究中,选择的毛霉属真菌在含有不同碳源的培养基中生长,并根据获得的光谱数据预测脂肪酸谱。FTIR光谱通过气相色谱 - 火焰离子化检测(GC - FD)进行脂肪酸分析校准。校准模型进行了交叉验证,得到的相关系数(R2)为0.71至0.78,均方根误差交叉验证值(RMSECV)为2.86%至6.96%(占总脂肪的百分比)。FTIR结果显示与GC分析结果有很强的相关性,其中在油菜籽、橄榄油和向日葵油中培养的铅灰毛霉VI02019以及在油菜籽和橄榄油中培养的冬生毛霉VI01993实现了高含量的不饱和脂肪酸(包括PUFA和MUFA)。