热带假丝酵母固体发酵代谢物提取方法的比较分析

A Comparison of Extraction Methods for Metabolites in Solid Fermentation with Candida tropicalis

本试验比较了几种提取微生物固体发酵代谢物的方法,旨在筛选出一种尽可能多保留了代谢物的方法。试验采用热带假丝酵母固体发酵棉粕源底物,以液相色谱串联质谱(LC-MS)为分析平台,分别采用50%冷甲醇溶液(甲醇法)、酸性甲醇-乙腈溶液(酸性甲醇-乙腈法)和50%乙腈溶液(乙腈法)提取发酵代谢物。结果表明:采用酸性甲醇-乙腈法、乙腈法和甲醇法分别提取出1 831、1 522和1 262种代谢物,提取效率表现为酸性甲醇-乙腈法>甲醇法>乙腈法。本试验中3种方法相比,酸性甲醇-乙腈法提取的代谢物种类最多,提取效率最佳,是微生物固体发酵代谢物提取的有效方法。

In order to find a good extraction method that can keep metabolites as many as possible,several extraction methods for solid fermentation metabolites were compared in this study.Substrates with cottonseed meal were fermented by Candida tropicalis,and the analysis of metabolites was carried out using the liquid chromatographic-mass spectrometry(LC-MS) platform.Three different extraction solutions,including 50% cold methanol solution(MeOH method),acidic methanol-acetonitrile solution(AMF method) and 50% acetonitrile solution(MeCN method),were used.The results showed that the number kinds of metabolites extracted by methods of AMF,MeCN and MeOH were 1 831,1 522 and 1 262,respectively;extraction efficiency was shown as AMF method>MeOH method>MeCN method.Among the three methods in this experiment,AMF method is an effective method for metabolites extraction in solid fermentation,and acts better on both kinds of metabolites and extraction efficiency.